Shaping of Hepatic Ischemia/Reperfusion Events: The Crucial Role of Mitochondria

  Citation: Teodoro, J.S.; Da Silva, R.T.; Machado, I.F.; Panisello-Roselló, A.; Roselló-Catafau, J.; Rolo, A.P.; Palmeira, C.M. Shaping of Hepatic Ischemia/Reperfusion Events: The Crucial Role of Mitochondria. Cells 2022,11, 688. https://doi.org/ 10.3390/cells11040688 Academic Editors: Anna-Liisa Nieminen and John J. Lemasters Received: 11 January 2022 Accepted: 11 February 2022 Published: 16 February 2022 Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). cells Review Shaping of Hepatic Ischemia/Reperfusion Events: The Crucial Role of Mitochondria João S. Teodoro 1,2,3,† , Rui T. Da Silva 2,4,† , Ivo F. Machado 2,3 , Arnau Panisello-Roselló4, Joan Roselló-Catafau 4, Anabela P. Rolo 1,2 and Carlos M. Palmeira 1,2,* 1MitoLab, Department of Life Sciences, University of Coimbra, 3000 Coimbra, Portugal; [email protected] (J.S.T.); [email protected] (A.P.R.) 2MitoLab, Mitochondria, Metabolism and Disease Group, Center for Neurosciences and Cell Biology, Faculdade de Medicina, University of Coimbra, 3000 Coimbra, Portugal; [email protected] (R.T.D.S.); [email protected] (I.F.M.) 3IIIUC–Institute of Interdisciplinary Research, University of Coimbra, Pólo II da Universidade de Coimbra, 3000 Coimbra, Portugal 4Experimental Pathology Department, Institute of Biomedical Research of Barcelona (IIBB), CSIC-IDIBAPS, 08036 Barcelona, Spain; [email protected] (A.P.-R.); [email protected] (J.R.-C.) *Correspondence: [email protected]; Tel.: +351-239-240-700 † These authors contributed equally to this work. Abstract: Hepatic ischemia reperfusion injury (HIRI) is a major hurdle in many clinical scenarios, including liver resection and transplantation. Various studies and countless surgical events have led to the observation of a strong correlation between HIRI induced by liver transplantation and early allograft-dysfunction development. The detrimental impact of HIRI has driven the pursuit of new ways to alleviate its adverse effects. At the core of HIRI lies mitochondrial dysfunction. Various studies, from both animal models and in clinical settings, have clearly shown that mitochondrial function is severely hampered by HIRI and that its preservation or restoration is a key indicator of successful organ recovery. Several strategies have been thus implemented throughout the years, targeting mitochondrial function. This work briefly discusses some the most utilized approaches, ranging from surgical practices to pharmacological interventions and highlights how novel strategies can be investigated and implemented by intricately discussing the way mitochondrial function is affected by HIRI. Keywords: mitochondria; liver; ischemia/reperfusion; liver surgery; conditioning 1. Introduction 1.1. The Liver The liver is an organ with dozens of functions in the body, ranging from the betterknown bile production to assist digestion, to others that are referenced less often, such as its involvement in carbohydrate (production and storage of glycogen; release of glucose to circulation; gluconeogenesis to generate glucose from amino acids, lactate or glycerol), protein (most circulating proteins are produced by the liver, as are most amino acids and some hormones such as angiotensinogen; hormones and other circulating proteins are broken down in the liver) or lipid (production of cholesterol, lipogenesis and triglyceride synthesis) metabolism. Other functions involve the detoxification of xenobiotics and some heavy metals, serving as a blood reservoir, producing lymph, storing vitamins and metallic ions and promoting blood immune activity (by the liver native Kupffer cells, as well as a panoply of other immune cells) [1,2]. Given of its range of roles, it is unsurprising that liver transplantation is one of the most commonly performed organ transplants. Liver injury and failure is an increasingly common event [ 3 ] and, despite improvements in survivability and transplant success, there is still a tremendous unbalance between liver donations and necessities. This is further Cells 2022,11, 688. https://doi.org/10.3390/cells11040688 https://www.mdpi.com/journal/cells Cells 2022,11, 688 2 of 24 aggravated by the fact that many (if not most) potential donors are barred from providing organs due to innate conditions, such as steatosis or steatohepatitis, cirrhosis, or cancer, just to name a few [ 4 ]. As such, novel therapeutic approaches and interventions that can increase both the pool of available donors as well as the restoration of homeostasis upon liver insult, both of which are urgently needed. One of the most common types of hepatic injury is that caused by the cut and eventual restoration of circulation, which is commonly known as ischemia and reperfusion. 1.2. Ischemia/Reperfusion Hepatic ischemia-reperfusion injury (HIRI) is an accumulation of processes and events that revolve around cellular and organelle damage upon blood-flow restriction (ischemia), which is followed by a seemingly contradictory augmentation of injury upon the restoration of blood flow (reperfusion) [ 5 ]. HIRI is the main reason for complications and even mortality in the setting of hepatic surgery or transplantation [ 6 , 7 ]. Interestingly, despite intense investigation on the matter, the exact causes of HIRI are still unclear [8]. There are two main types of HIRI, depending on the setting of the organ at the time of ischemia, i.e., whether it is still inside the organism (warm) or outside, as is the case of transplantation (cold) [ 8 , 9 ]. Although in different induction settings, the pathophysiological events that take place are quite similar between the two types of HIRI [ 9 ]. While warm (normothermic, 22–25 ◦ C) HIRI begins with hepatocyte injury and during different events such as trauma, surgery or other events that restrict blood flow, cold (ice temperature, i.e., 0–1 ◦C ) HIRI only applies to a transplant setting, where the liver is harvested and transported in quasi-freezing temperatures in order to reduce tissue degradation. Regardless, both events result in an immune response triggered not by pathogens per se, but due to the release of pro-inflammatory signals [10], in what is known as sterile inflammation. There are many processes involved in HIRI events, namely the metabolic shift towards an anaerobic metabolism, mitochondrial dysfunction and oxidative stress (i.e., overproduction of reactive oxygen species that overbear natural antioxidant defenses such as scavenger molecules and, more relevantly, antioxidative enzymes, which contributes to alterations in redox signaling and molecular injury [ 11 ]), calcium overload and the immune response, heavily mediated by Kupffer cell and other immune cell types’ activation, such as infiltrating neutrophils and macrophages [ 12 , 13 ]. As such, given the central role of the liver in metabolic and energetic whole-body homeostasis, it comes as no surprise that HIRI causes a broad-range effect on the body. In fact, during the ischemic period, the lack of oxygen shifts the ATP-generating processes from aerobic respiration (i.e., oxidative phosphorylation, OXPHOS) towards anaerobic respiration, or glycolysis. However, given enough time, the lack of oxygen results in a shutdown of redox processes, severe reduction in ATP generation capacity, and parallel acidification of the cellular milieu, given the accumulation of lactic acid and ketone bodies, resulting in what is known as metabolic acidosis. Thus, the lower pH results in enzyme, organelle and even cellular injury [ 14 ]. As expected, once blood flow is restored (reperfusion), acidification is neutralized, which is paradoxically responsible for further injury due to the activation of pH-dependent proteases and phospholipases [ 15 , 16 ]. In parallel, the low O 2 pressure can also result in the elevation of cyclic AMP (cAMP) levels, resulting in the activation of cAMP-sensitive enzymes, with concomitant phosphorylation and perturbation of the function of key enzymes of the carbohydrate metabolism [ 17 ], which further contributes to the accumulation of acidic metabolites [8]. 1.3. Mitochondrial Function and HIRI Mitochondria are the essential players in the metabolism of eukaryotes. Virtually all of the ATP requirements of the nucleated cell derive from the processes taking place within mitochondria, i.e., the citric acid (Krebs) cycle and OXPHOS. While the Krebs cycle takes place exclusively on the mitochondrial matrix, taking in Acetyl-CoA to generate reducing equivalents (NADH and FADH 2 ), OXPHOS is a process that takes place mostly within the inner mitochondrial matrix (apart from the cytochrome celectron transport Cells 2022,11, 688 3 of 24 in the intermembrane space), where electrons provided by the aforementioned reducing equivalents are transported in energetically favorable leaps along the respiratory chain (from Complexes I and II towards III and then IV) towards a molecule of oxygen, generating water. In tandem with the energetically favorable electronic transport, there is vectorial, the energetically unfavorable (against the concentration gradient) transport of protons from the matrix towards the intermembrane space, thus traversing the mostly protonimpermeable inner membrane. This generates an electrochemical gradient across this membrane (electrical due to the charge disparity, since protons are charged; chemical since protons are what make a solution acid), which is the storage of a tremendous potential energy. This energy is used to create a covalent bond between adenine diphosphate (ADP) and a molecule of ionic phosphate (Pi), generating adenine triphosphate (ATP), the cell’s energetic currency. This phosphorylation of ADP takes place at the level of the ATP synthase or complex V, a protonic channel bound to a catalytic head [18]. However, given that OXPHOS heavily relies on electron transport in a biological setting, it is thus expected that some instability is present. In fact, most of the typical cell’s reactive oxygen species (ROS) are produced in mitochondria simply as by-products of cellular respiration [ 19 ]. First considered as unwanted by-products of mitochondrial ATP generation, ROS are now ubiquitously understood as necessary, given their production does not exceed manageable levels, since mild oxidative stress is a fundamental modulator of redox signaling and the maintenance of adequate defenses and function [ 20 ]. However, it is true that excessive, prolonged ROS generation invariably results in oxidative stress, causing impaired mitochondrial function and exacerbated ROS generation, creating a snowball effect of oxidative stress that ultimately might lead to the cell’s death. Mitochondria are particularly susceptible to oxidative stress, since several of its components are severely damaged by ROS, such as mitochondrial DNA (mtDNA), which are phospholipids from the membranes or proteic elements of various metabolic pathways. For example, thiol groups within Complex I of the respiratory chain are readily oxidized, resulting in elevated ROS generation due to the mishandling of electron transport [ 21 ]. Furthermore, cardiolipin, a hallmark phospholipid of the inner mitochondrial membrane and the element most responsible for this membrane’s protonic impermeability (and, as such, for the membrane potential), is composed of highly unsaturated fatty acids, which are also prime targets for oxidation [ 22 ]. Furthermore, cardiolipin also has a regulatory role in the function of various enzymes, such as creatine kinase [ 23 ]. Metabolism is also obviously affected, and ß-oxidation is particularly susceptible, since increased acylation of proteins due to elevated matrix accumulation of acyl-CoA was found in ischemia/reperfusion events [24,25] . Another metabolic consequence is the depletion of NAD + , an essential co-factor for numerous metabolic pathways (such as glycolysis and Krebs cycle, just to name a few) and important enzymes such as NAD + -dependent sirtuins, deacetylases involved in cellular survival. Since OXPHOS is the major syphon for NADH, refreshing the NAD + pool, the drastic reduction in NADH consumption helps in this pool’s exhaustion [25,26]. Paradoxically, hypoxic conditions appear to create a prime environment for ROS generation, since not all of the oxygen supply is removed, but the shift towards an anaerobic metabolism is a driver for ROS generation, particularly hydrogen peroxide [ 27 ]. Figure 1 illustrates this escalation of injury, where damage to mitochondrial function and integrity escalates to tissue damage and, eventually, to organ failure. Cells 2022,11, 688 4 of 24 Cells 2022, 11, x FOR PEER REVIEW 4 of 25 Figure 1. The upscaling of damage in HIRI. Severe compromise to mitochondrial function results in the exacerbated generation of reactive oxygen species (ROS), resulting in the activation of pro-apoptotic protocols such as the opening of the mitochondrial permeability transition pore (mPTP) and release of pro-apoptotic factors, such as ionic calcium (Ca 2+ ). While mitochondrial Ca 2+ release is markedly lower when compared with other Ca 2+ sources that could lead to elevated levels (for example, the endoplasmic reticulum or from extracellular sources), the damage to mitochondrial function and integrity will undoubtedly lead to the release of Ca 2+ and other pro-apoptotic factors. Furthermore, these sources could initiate mitochondrial dysfunction, rather than mitochondrial injury per se. Regardless, given enough replication of these phenomena, cellular survival is at risk, which in turn is a marker for further damage, due to the activation of sterile inflammatory procedures. All these processes, if left unchecked, might result in tissue loss and, eventually, organ failure. One way by which the cellular mitochondrial population responds and adapts to fluctuating metabolic and biophysics conditions is through the modulation of mitochondrial dynamics, i.e., through alterations in mitochondrial fusion, fission, degradation and biogenesis events. These alterations result in the modification of the mitochondrial bioenergetic capacity, not only through elevated OXPHOS elements’ production, but also by more macro alterations, such as reticulation and volume [28,29]. In typical conditions, the mitochondrial network within a cell is found to be highly regulated to serve the cell’s needs (for example, mitochondria are typically found in great numbers around the nucleus). However, if an elevated energetic output is necessary, mitochondria usually undertake fission and biogenesis protocols, to elevate organelles’ numbers. In tandem, the fission process also allows for the isolation and removal, by intracellular degradation, of incompetent or damaged mitochondrial units or elements, in order to boost energetic production by the mitochondrial unit [30]. Thus, mitochondrial dynamics (i.e., the fluid shape, size and numbers) are paramount to a proper and efficient response to the evershifting metabolic environment and to reply to the sometimes quite different cellular necessities. Of course, all metabolic effects deeply involve mitochondria, whereby virtually all of the cell’s ATP needs are produced, given that oxygen is available. Concomitantly, the low O 2 pressure in ischemia all but impedes ATP generation by OXPHOS, which has immediate effects on various cellular processes, such as ion balance which, in turn, help the loss of mitochondrial membrane potential, the driving force for ATP generation in aerobiosis [15,31]. This in turn leads to the induction of what is known as the permeability transition (mPT), where smaller than 1.6 kDa (in high-conductance state) or up to 0.3 kDa (in lowconductance state) solutes can freely cross the mitochondrial membranes, including various pro-apoptotic factors [32,33], in the apparent formation of unspecific pores of dubious, possibly fluid, composition [34]. As expected, low O2 pressure results in a decreased mitochondrial biogenesis, as hypoxia-sensitive elements such as hypoxia-inducible factors are negative regulators of the peroxisome proliferator activated receptor gamma co-activator 1 alpha (PGC-1α), the Figure 1. The upscaling of damage in HIRI. Severe compromise to mitochondrial function results in the exacerbated generation of reactive oxygen species (ROS), resulting in the activation of proapoptotic protocols such as the opening of the mitochondrial permeability transition pore (mPTP) and release of pro-apoptotic factors, such as ionic calcium (Ca 2+ ). While mitochondrial Ca 2+ release is markedly lower when compared with other Ca 2+ sources that could lead to elevated levels (for example, the endoplasmic reticulum or from extracellular sources), the damage to mitochondrial function and integrity will undoubtedly lead to the release of Ca 2+ and other pro-apoptotic factors. Furthermore, these sources could initiate mitochondrial dysfunction, rather than mitochondrial injury per se. Regardless, given enough replication of these phenomena, cellular survival is at risk, which in turn is a marker for further damage, due to the activation of sterile inflammatory procedures. All these processes, if left unchecked, might result in tissue loss and, eventually, organ failure. One way by which the cellular mitochondrial population responds and adapts to fluctuating metabolic and biophysics conditions is through the modulation of mitochondrial dynamics, i.e., through alterations in mitochondrial fusion, fission, degradation and biogenesis events. These alterations result in the modification of the mitochondrial bioenergetic capacity, not only through elevated OXPHOS elements’ production, but also by more macro alterations, such as reticulation and volume [ 28 , 29 ]. In typical conditions, the mitochondrial network within a cell is found to be highly regulated to serve the cell’s needs (for example, mitochondria are typically found in great numbers around the nucleus). However, if an elevated energetic output is necessary, mitochondria usually undertake fission and biogenesis protocols, to elevate organelles’ numbers. In tandem, the fission process also allows for the isolation and removal, by intracellular degradation, of incompetent or damaged mitochondrial units or elements, in order to boost energetic production by the mitochondrial unit [ 30 ]. Thus, mitochondrial dynamics (i.e., the fluid shape, size and numbers) are paramount to a proper and efficient response to the ever-shifting metabolic environment and to reply to the sometimes quite different cellular necessities. Of course, all metabolic effects deeply involve mitochondria, whereby virtually all of the cell’s ATP needs are produced, given that oxygen is available. Concomitantly, the low O 2 pressure in ischemia all but impedes ATP generation by OXPHOS, which has immediate effects on various cellular processes, such as ion balance which, in turn, help the loss of mitochondrial membrane potential, the driving force for ATP generation in aerobiosis [ 15 , 31 ]. This in turn leads to the induction of what is known as the permeability transition (mPT), where smaller than 1.6 kDa (in high-conductance state) or up to 0.3 kDa (in low-conductance state) solutes can freely cross the mitochondrial membranes, including various pro-apoptotic factors [ 32 , 33 ], in the apparent formation of unspecific pores of dubious, possibly fluid, composition [34]. As expected, low O 2 pressure results in a decreased mitochondrial biogenesis, as hypoxia-sensitive elements such as hypoxia-inducible factors are negative regulators of the peroxisome proliferator activated receptor gamma co-activator 1 alpha (PGC-1 α ), the master regulator of mitochondrial biogenesis [ 35 ]. In a similar fashion, the other dynamism Cells 2022,11, 688 5 of 24 fluxes are also perturbed. At the start of reperfusion, there is a drive for increased fission, due to elevated levels of dynamin-related protein 1 (Drp1) acting in harmony with other pro-apoptotic proteins such as Bax or Bak [ 36 ], a mechanism that appears to be heavily related to altered calcium metabolism (Ca 2+ flux perturbation are discussed in more detail below). Conversely, Mitofusin protein 1 (Mfn1) and the optic atrophy protein 1 (Opa1), which are involved in the mitochondrial fusion processes, are downregulated by oxidative stress in ischemic events [ 37 , 38 ]. As for mitophagy, it is now ubiquitously accepted that this process is severely limited in HIRI, and that addressing it yields protection against pathogenicity [36,39,40]. As previously mentioned, one of the most affected ionic balances is the one pertaining to calcium (Ca 2+ ), a ubiquitous and powerful secondary messenger as well as a cofactor for various enzymes (including some involved in cellular processes of self-degradation). As expected, given these roles, its intracellular levels are highly regulated [ 41 ]. Along with the endoplasmic reticulum, mitochondria are the main site of Ca 2+ storage for a quick and localized release upon specific signaling processes, typically by using the mitochondrial membrane potential ( ∆Ψ ) as a token for Ca 2+ accumulation against the gradient [ 42 ]. In ischemia, cytosolic Ca 2+ levels increase (partly due to the diminished ∆Ψ , as mentioned above), resulting in the activation of Ca 2+ -sensitive enzymes (ex: calpains, calmodulin, protein kinase C, amongst many others), elevated oxidative stress and other deleterious events that typically result in cellular death by apoptosis [43,44]. Cellular death usually leads to the release, to the extracellular milieu, of pro-inflammatory factors that induce an immune response and overall localized aggravation of damage. In the liver, at the start (i.e., during hypoxia), Kupffer cells induce zonal injury via a combination of elevated ROS generation (generated primarily by white blood cells that are chemoattracted to the site such as polymorphonuclear neutrophils [ 45 ]) and release of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNFα ) and interleukin1beta (IL-1 β ). These, in turn, lead to the attraction, migration, adhesion and chemotaxis of neutrophils, exacerbating the immune activity in the area [ 46 , 47 ]. However, this process further escalates, particularly after blood-flow restoration (reperfusion), partly due to the established organelle injury suffered, but also caused by the already undergoing cellular injury processes [ 48 ]. Of course, if mitochondrial function is compromised by these processes, a further escalation of injury is sure to ensue, and that is why the conservation of mitochondrial capacity and activity might tip the scale towards the survival of the cell against HIRI. As an example of the responsibility of mitochondria in this survival is their aforementioned role in Ca 2+ handling. As mentioned, mitochondria are not the only reservoir for Ca 2+ storage, but simultaneously are one of the most important intracellular calcium storage locations, due to an exchange with membrane potential [ 49 ]. As such, the quick and efficient removal of Ca 2+ from the cytosolic milieu is literally vital for the cell and, as such, mitochondria in prime conditions are mandatory for this effect, not just because of the membrane potential swap, but also because of the effects of Ca 2+ within mitochondria. In fact, this ion’s presence in the organelle’s matrix is a major event in OXPHOS and ATP generation, since excess Ca 2+ can oftentimes lead to mitochondrial rupture and the release of pro-apoptotic signals [ 49 ]. As such, mitochondria are a paramount player in intracellular Ca 2+ metabolism, which is particularly relevant in non-excitable tissues that have poor endoplasmic reticulum (ER) Ca 2+ clearance capacity and, since mitochondrial Ca 2+ handling appears to be crucial not in homeostasis, but in situations of stress such as HIRI [ 42 ] further implicates mitochondrial function conservation as paramount for the successful recovery from HIRI. Unsurprisingly, mitochondria can be found in high numbers near Ca2+ release points, as to better control local ionic concentration, guaranteeing a localized Ca 2+ effect, rather than a full cellular response, which is typically associated with events of apoptosis [49,50]. Finally, it has also been shown that mitochondria play a role in immune responses, being a central element in the metabolic transition of immune cells in proliferation as well as Cells 2022,11, 688 6 of 24 in inflammatory signaling [ 51 ]. Since the inflammatory process typically requires sustained oxidative stress, it is unsurprising to consider that mitochondria are considered part of the process of inflammation. In fact, their role in mediating and, in some cases, initiating the inflammatory process, was completely unknown until very recently. Recent works have shown that mitochondria actively release some of its DNA molecules (mtDNA), which are powerful pro-inflammatory elements [ 52 – 54 ]. A reason for the apparent recognition of mtDNA as an antigen might rely on the mitochondrial bacterial origin, for mtDNA is markedly over methylated when compared with nuclear DNA [ 55 ]. Furthermore, unlike the commonly depicted bacterial-like circular DNA molecule, mtDNA is in fact tightly packed with proteins, one of which is the mitochondrial transcription factor A (TFAM), which various studies have identified as a potent immunostimulator [ 56 , 57 ]. Curiously, there also appears to be a role for oxidative stress in this matter, as only mtDNA with oxidized bases was capable of eliciting an immune response [ 58 ]. There are many other ways by which mtDNA can be a pro-inflammatory molecule, and this topic has been excellently revised in previous research [52,55]. NF-kB, a known pro-inflammatory cytokine, has also been shown to be activated by the loss of mitochondrial membrane integrity [ 52 ]. The NOD-, LRR-, or pyrin domaincontaining protein 3 (NLRP3) inflammasome proteins are ROS-dependently localized to mitochondria [ 59 , 60 ]. In tandem, mitochondria also have several molecules recognized by the NLRP3 inflammasome, such as cardolipin or even the mitochondrial DNA [ 60 – 62 ]. Mitochondria further enhance this inflammasome’s activity by the release of various activators (such as, for instance, cytochrome cor Smac), causing in turn the release of proinflammatory cytokines IL-1ß and IL-18 [ 60 ]. These pathways are apparently carefully preserved in various species, since they are fundamental following bacterial or viral infections [ 61 , 63 ]. However, this should be finely balanced, since these processes are also linked to highly inflammatory processes in pathologic conditions, such as HIRI [ 64 ]. There are also numerous evidences of mitochondrial roles in cancer and neurodegeneratory immunological responses (which have been extensively reviewed before [65]). In summary, since mitochondria have a vital role in a panoply of cellular activities, it is easy to understand how mitochondrial dysfunction is intimately associated with pathological processes. As such, maintaining or restoring mitochondrial homeostasis is a widely studied and desired goal for countering various diseases, such as HIRI. 2. Surgical Approaches in HIRI Mitochondrial Function Compromise In order to achieve the preservation of mitochondrial function, various surgical practices have been tested and utilized in a clinical setting for over a decade. They mostly rely on a form of damage priming, i.e., the introduction of small bouts of damage through mild, short, controlled events of removal and restoration of blood flow, in a manner that is now recognized as hormetic [ 30 ], a concept and phenomenon discussed further down. We discuss the most common ones, explaining their similarities and differences, and how they relate to mitochondrial function preservation. 2.1. Ischemic Preconditioning Ischemic preconditioning (IPC) is a strategy that has shown promising results in the bench. To achieve hepatic IPC, a clamp or other means of blood flow restriction is used for a small period of time (usually 10–15 min) after which flow is restored by restrictor removal. After a proportional amount of time has passed, the procedure that will generate HIRI is then initiated [ 66 ]. While no definitive answers exist as to why IPC is a protective strategy against HIRI, some elements have been confirmed to be crucial, such as ATP levels and, thus, mitochondrial function integrity [ 67 – 70 ]. In tandem with preserved mitochondrial function and integrity, elevated mitophagy [ 71 , 72 ] and prevention of apoptosis are also key events [ 73 , 74 ]. In fact, not just mitophagy is important, but apparently also other cellular components’ replacement is a part of the process [ 75 , 76 ]. However, some criticism about the efficiency of this approach exists, because some meta-analyses have concluded that IPC Cells 2022,11, 688 7 of 24 is not always a guarantee of improved HIRI, since there is simply too much heterogeneity in the human population or, more clearly, in the patients studied [39,69]. It is impossible though to discuss IPC without the introduction of the concept of hormesis and, more specifically, mitohormesis. While previous work has discussed in great detail the intricacies of mitohormesis [ 30 ], it should suffice, for this works’ purpose, to acknowledge that the mitohormetic process is one where a small injurious event leads to the fairly limited but definitely present injury in mitochondria, in particular in subsets or even subsections of the organelles. These are quickly identified and removed through various mitodynamic processes such as fission and mitophagy, and are replaced by newer, more competent units, thus contributing to a more resilient cell. This is far from a novel concept or even observation, as various reports on the matter have surfaced for at least two decades, involving both intracellular events [ 77 , 78 ] or even the liver’s immune system [ 79 ], which is apparently also a critical participant. 2.2. Intermittent Clamping A similar surgical procedure to IPC is known as intermittent clamping (IC). The main difference is the number of occlusion-flow restoration cycles and the timing of performance; whereby IPC is by default just one cycle before the HIRI event, IC consists of various cycles of flow occlusion and restoration, and these are not limited to just the pre-HIRI event. In fact, IC can be spread, even during the HIRI event [ 66 ]. In terms of hepatic HIRI, IC has been shown to be superior [ 80 ], inferior [ 81 ] and even similar to IPC [ 67 ]. These differences might fall prey to various causes, of which differences in intervention time is but one of them (different populations intervened, different medical backgrounds, etc.). However, surgery timings are possibly of the most important, for the authors of various studies have concluded that, for shorter interventions, IPC is superior as it improves transaminase levels and surgical complications but, for longer time frames, they are virtually the same procedure [ 60 , 63 ], for which mitochondrial function preservation and cellular ATP levels’ maintenance are imperative [82]. A variation of this procedure is ischemic post-conditioning (IPostC), where the bouts of flow restriction/restoration are performed after the main ischemic event, so it acts more like an IC but between the phases of ischemia and reperfusion [ 83 , 84 ]. This method results in virtually the same results as pre-HIRI IC, albeit the lack of time-consuming flow restriction/restoration events to initiate the surgical procedure might be advantageous in certain settings where there is a rush to initiate the surgical event, such as, for instance, transplantation [85,86]. 2.3. Remote Ischemic Preconditioning Another surgical procedure to tackle HIRI is known as remote IPC (RIPC). This is a rather bizarre phenomenon whereby events of ischemia/reperfusion localized in distant organs results in improved liver resistance to HIRI. While much is yet to be understood about this phenomenon, it is undoubtedly a reality, as many works have reported quasiunbelievable responses of the liver (and other organs as well) to remote IPC events. While the release of protective elements (what those are is still a hotly debated topic) is probably involved and can flow in the circulation, protecting distant tissues, mitochondrial-function protection in the remote tissue/organ is apparently a necessity for RIPC in the liver [ 87 – 91 ]. 2.4. Machine Perfusion A technique that has attracted much attention due to its potential for use in a particular setting (transplantation of less-than-optimal tissue samples) is hypothermic machine perfusion, HMP [ 92 ]. This method’s intents are to allow for these otherwise rejected organ donations to be reconditioned and to achieve an extension of the preservation time window [ 86 ]. In this technique, the organ to be transplanted is perfused in a similar solution to the one used in normal, static cold (4 ◦ C) storage, but with a faster flow. However, for obvious reasons, function testing is not possible, since the idea is to aggressively lower Cells 2022,11, 688 8 of 24 the organ’s temperature. A key modification to this protocol is to oxygenize the perfusion solution, which appears to prevent HIRI in these organs, since ROS generation and inflammation are reduced, while the ATP-generation capacity of mitochondria is salvaged [ 93 ]. While normothermic (i.e., core temperature) perfusion is both the more common method and results in lower levels of HIRI, allowing for increased graft viability (up to around 19 h), HMP has the significant advantage to help expand the pool of viable donors to otherwise unusable and certain immunorejection grafts [94]. 2.5. Mitochondrial Transplantation Finally, although it is not a surgical technique as classically defined, the transplantation of mitochondria is a surprisingly effective approach [ 95 , 96 ]. In fact, in HIRI studies, mitochondrial transplantation resulted in improved serum transaminase levels, decreased inflammatory markers and other success indicators, after mitochondria were injected directly into the spleen, from where they migrated towards the injured liver [ 94 , 97 ]. However, there are still many questions regarding this novel approach. It is not clear how mitochondria migrate to, enter the injured cell and start working to replace the damaged ones, but further studies on the matter will certainly help explain this matter. 3. Pharmacological Intervention for HIRI It is well established that the mechanisms involved in HIRI are of a multifactorial nature, which entails complex signaling pathways. The modulation of protective/deleterious pathways using pharmacological interventions represents an attractive approach in this context, since it is theoretically cheaper and safer than surgical approaches. Pharmacological preconditioning relies on the administration of specific drugs mimicking the protective mechanisms and biologic effects of IPC. Although the exact mechanism is still not fully understood, it seems that the protection can be achieved, at least in part, by the modulation of well characterized pathways, namely the Reperfusion Injury Salvage Kinase (RISK), the Survivor Activating Factor Enhancement (SAFE), the cyclic guanosine 3 0 ,5 0 -monophosphate/ Protein Kinase G (cGMP/PKG) as well as a combination of others, including inflammatory, metabolic and mitochondrial factors (for an excellent revision on this matter, please consult [98]). 3.1. Mitochondrial Targeting for HIRI 3.1.1. Natural Compounds The vast majority of the pharmacological interventions involve the administration of natural (i.e., melatonin [ 99 ]), mimetic (i.e., N-acetylcysteine, NAC [ 100 ]), or metabolism shifting agents (i.e., trimetazidine [ 101 ] or indirubin-3 0 -oxime [ 102 ]). Several studies have shown the potential role of melatonin in reducing IRI. Zhang and colleagues reported that melatonin diminished myocardial IRI through the improvement of mitochondrial fusion and mitophagy, as well as the activation of AMPK-OPA1 signaling pathways [103]. Melatonin was also found to decrease IRI via the upregulation of the mitochondrial sirtuin 3 and a subsequent reduction in oxidative stress and apoptosis [ 104 ]. In addition, melatonin conditioning also alleviated hepatic IRI via the suppression of the induction of mPT [105]. Based on the crucial role of ROS in HIRI pathophysiology, pharmacological interventions aiming to neutralize or modulate its production using antioxidants were one of the first pharmacological approaches attempted. NAC is a precursor of the synthesis of glutathione, the main endogenous ROS scavenger involved in the cellular protection against oxidative stress, as well as directly scavenging pro-oxidant agents with unpaired electrons [ 100 ]. Numerous studies have reported the hepatoprotective effects of NAC administration prior to HIRI, especially via the significant reduction of transaminase release [ 106 ] and decreased oxidative stress, resulting in diminished apoptosis and autophagy [107,108]. Trimetazidine (TMZ) is a piperazine derivative that has been used as an anti-ischemic agent. The prevention of ROS production, mitochondrial damage (such as protein and phospholipid oxidation, to name a few) and decreased ATP levels are thought to be im- Cells 2022,11, 688 9 of 24 portant targets by which TMZ exerts its cytoprotective effect [ 109 ], for while TMZ is not an antioxidant per se, its action in metabolic modulation results in elevated antioxidant enzyme activity [ 110 ]. The presence of TMZ confers a reduction in hepatic injury and an improved fatty liver functionality after ischemia/reperfusion, through the activation of AMPK and subsequent increase of nitric oxide levels [111]. The pharmacological preconditioning with indirubin-3 0 -oxime was demonstrated to protect the liver against HIRI by preserving mitochondrial function and hepatic energetic balance [ 112 ]. This compound inhibits GSK-3 β and, consequently, prevents cyclophilin D phosphorylation by GSK-3 β . Since cyclophilin D serine residues’ phosphorylation leads to mPT induction [ 112 , 113 ], its inhibition modulates the susceptibility to mPT induction, thus preserving mitochondrial function following HIRI. 3.1.2. Synthetic and Directed Agents OXPHOS Elements Manipulation Succinate accumulation has been heavily implied in HIRI [ 114 ]. By using succinate dehydrogenase (OXPHOS Complex II) inhibitors such as malonate, succinate accumulation is prevented and thus ischemic injury is reduced [ 114 ]. Similarly, Complex I inhibitors have been investigated. It is true that a wide panoply of mitochondrial-beneficial agents have Complex I inhibitory capacity (ex: Metformin, Berberine, to name a few) but those appear to rely on mitohormetic effects; here, other types of molecules such as MitoSNO (mitochondrial-specific S-nitrosating agent) or Amobarbital have proven protection in ischemia/reperfusion [ 115 , 116 ], effects that might depend on diminished pro-apoptotic factors’ release for the prevention of retrograde electron flow and thus increased ROS generation [36]. Mitochondrially Targeted Antioxidants Unlike the aforementioned NAC, which has a wider range of activity than just mitochondria, other antioxidants specifically designed to target mitochondria have already been tested for the same reasons. For instance, Coenzyme Q 10 (or ubiquinol) is not only a member of the respiratory chain but also a potent antioxidant, which has proven to be able to prevent HIRI [ 117 , 118 ]. MitoQ and SkQ1 are other mitochondrial-specific antioxidant molecules, modelled after the widely known tetraphenylphosphonium (TPP + ), a molecule that freely traverses the inner mitochondrial membrane and, as such, this property was explored by various researchers to deliver antioxidant agents to the mitochondrial matrix [ 119 , 120 ]. Other classes of compounds, with different antioxidant roots and delivery alternatives exist and most have shown similarly effective results, such as Bendavia (a Szeto-Schiller peptide [ 121 ]), MitoGSH [ 122 ], or Euk-8 [ 123 ] although not all exactly on HIRI, but on other tissues/organs, which opens the possibility for more studies. NAD+Metabolism As previously mentioned, NAD + is an essential co-factor to many metabolic reactions, and in low abundance in HIRI. To further complicate matters, NAD + transport across biological membranes is a very complicated procedure, and oftentimes impossible although in recent years some transporters have been identified [ 124 ]. As such, most studies have focused on more mobile NAD+ biosynthesis precursors nicotinamide riboside (NR) and nicotinamide mononucleotide (NMN). Their use boosts the cell’s NAD + pool, a feature already explored in various works [125,126]. PPAR Agonists Another strategy that is not immediately intuitive is the use of agonists of the nuclear receptor family of peroxisome proliferator-activated receptor. For instance, fibrates that directly activate PPAR α , a commonly used antidiabetic class of drugs, have been shown to attenuate HIRI [ 127 ]. Even more striking is the fact that the PPAR γ agonist pioglitazone Cells 2022,11, 688 16 of 24 5. Concluding Remarks It is now clear that many of the cellular and mitochondrial events in HIRI are also present in other organs/tissues subjected to a similar event. This is extremely encouraging and helpful, as it means that virtually all of the research conducted in this field of ischemia/reperfusion is highly transversal between organs, saving both time and resources [ 98 ]. From all of the explored literature, patterns of intervention begin to emerge, which have been mentioned in this work. Most strategies would thus benefit from a combination of the main goals of the works discussed here, ranging from reduced oxidative stress from the earliest possible time to the elevation of mitochondrial numbers, activity and resilience. Author Contributions: All the authors state that this work is original and of their own making and was not published before and/or elsewhere, nor is in consideration in another journal. All authors contributed equitably to this manuscript. Writing: J.S.T., R.T.D.S., I.F.M., A.P.-R.; Revision: J.S.T., J.R.-C., A.P.R. and C.M.P. All authors have read and agreed to the published version of the manuscript. Funding: This work was financed by the European Regional Development Fund (ERDF), through the COMPETE 2020—Operational Programme for Competitiveness and Internationalization and Portuguese National Funds via FCT—Fundação para a Ciência e a Tecnologia, under project UIDB/04539/ 2020 and UIDP/04539/2020. JST is recipient of a CEEC researcher grant from FCT and CNC (CEECIND/4400/2007). IFM is recipient of a PhD scholarship from FCT (DFA/BD/8529/2020). RTS was a recipient of a Foie Gras Early Research Training Grant, from the European Union’s Horizon 2020 Research and Innovation programme under the Marie Sklodowska-Curie Grant (Agreement No. 722619). Conflicts of Interest: The authors declare no conflict of interest with the publication of this work. References 1. Kalra, A.; Tuma, F. Physiology, Liver; StatPearls Publishing: Treasure Island, FL, USA, 2018. 2. Rui, L. Energy Metabolism in the Liver. Compr. Physiol. 2014,4, 177–197. [CrossRef] [PubMed] 3. Bodzin, A.S.; Baker, T.B. Liver Transplantation Today: Where We Are Now and Where We Are Going. Liver Transplant. 2018 ,24, 1470–1475. [CrossRef] [PubMed] 4. Ghinolfi, D.; Melandro, F.; Torri, F.; Martinelli, C.; Cappello, V.; Babboni, S.; Silvestrini, B.; de Simone, P.; Basta, G.; del Turco, S. Extended Criteria Grafts and Emerging Therapeutics Strategy in Liver Transplantation. The Unstable Balance between Damage and Repair. Transplant. Rev. 2021,35, 100639–100649. [CrossRef] 5. Lee, S.H.; Culberson, C.; Korneszczuk, K.; Clemens, M.G. Differential Mechanisms of Hepatic Vascular Dysregulation with Mild vs. Moderate Ischemia-Reperfusion. Am. J. Physiol. Gastrointest. Liver Physiol. 2008,294, G1219–G1226. [CrossRef] 6. Sheng, M.; Zhou, Y.; Yu, W.; Weng, Y.; Xu, R.; Du, H. Protective Effect of Berberine Pretreatment in Hepatic Ischemia/Reperfusion Injury of Rat. Transplant. Proc. 2015,47, 275–282. [CrossRef] 7. Nardo, B.; Bertelli, R.; Montalti, R.; Beltempo, P.; Puviani, L.; Pacilè, V.; Cavallari, A. Preliminary Results of a Clinical Randomized Study Comparing Celsior and HTK Solutions in Liver Preservation for Transplantation. Transplant. Proc. 2005 ,37, 320–322. [CrossRef] 8. Cannistrà, M.; Ruggiero, M.; Zullo, A.; Gallelli, G.; Serafini, S.; Maria, M.; Naso, A.; Grande, R.; Serra, R.; Nardo, B. Hepatic Ischemia Reperfusion Injury: A Systematic Review of Literature and the Role of Current Drugs and Biomarkers. Int. J. Surg. 2016 , 33, S57–S70. [CrossRef] 9. Zhai, Y.; Petrowsky, H.; Hong, J.C.; Busuttil, R.W.; Kupiec-Weglinski, J.W. Ischaemia-Reperfusion Injury in Liver Transplantationfrom Bench to Bedside. Nat. Rev. Gastroenterol. Hepatol. 2013,10, 79–89. [CrossRef] 10. Zhai, Y.; Busuttil, R.W.; Kupiec-Weglinski, J.W. Liver Ischemia and Reperfusion Injury: New Insights into Mechanisms of Innate-Adaptive Immune-Mediated Tissue Inflammation. Am. J. Transplant. 2011,11, 1563–1569. [CrossRef] 11. Sies, H. Oxidative Stress: A Concept in Redox Biology and Medicine. Redox Biol. 2015,4, 180–183. [CrossRef] 12. Weigand, K.; Brost, S.; Steinebrunner, N.; Bchler, M.; Schemmer, P.; Müller, M. Ischemia/Reperfusion Injury in Liver Surgery and Transplantation: Pathophysiology. HPB Surg. 2012,2012, 176723–176731. [CrossRef] [PubMed] 13. Montalvo-Jave, E.E.; Escalante-Tattersfield, T.; Ortega-Salgado, J.A.; Piña, E.; Geller, D.A. Factors in the Pathophysiology of the Liver Ischemia-Reperfusion Injury. J. Surg. Res. 2008,147, 153–159. [CrossRef] [PubMed] 14. Siriussawakul, A.; Zaky, A.; Lang, J.D. Role of Nitric Oxide in Hepatic Ischemia-Reperfusion Injury. World J. Gastroenterol. 2010 , 16, 6079–6086. [CrossRef] [PubMed] 15. Guan, L.-Y.; Fu, P.-Y.; Li, P.-D.; Li, Z.-N.; Liu, H.-Y.; Xin, M.-G.; Li, W. Mechanisms of Hepatic Ischemia-Reperfusion Injury and Protective Effects of Nitric Oxide. World J. Gastrointest. Surg. 2014,6, 122. [CrossRef] Cells 2022,11, 688 17 of 24 16. Datta, G.; Fuller, B.J.; Davidson, B.R. Molecular Mechanisms of Liver Ischemia Reperfusion Injury: Insights from Transgenic Knockout Models. World J. Gastroenterol. 2013,19, 1683–1698. [CrossRef] 17. Guan, Y.-F. Ischemic Post-Conditioning to Counteract Intestinal Ischemia/Reperfusion Injury. World J. Gastrointest. Pathophysiol. 2010,1, 137–143. [CrossRef] 18. Nolfi-Donegan, D.; Braganza, A.; Shiva, S. Mitochondrial Electron Transport Chain: Oxidative Phosphorylation, Oxidant Production, and Methods of Measurement. Redox Biol. 2020,37, 101674–101683. [CrossRef] 19. Kasai, S.; Shimizu, S.; Tatara, Y.; Mimura, J.; Itoh, K. Regulation of Nrf2 by Mitochondrial Reactive Oxygen Species in Physiology and Pathology. Biomolecules 2020,10, 320. [CrossRef] 20. Shields, H.J.; Traa, A.; van Raamsdonk, J.M. Beneficial and Detrimental Effects of Reactive Oxygen Species on Lifespan: A Comprehensive Review of Comparative and Experimental Studies. Front. Cell Dev. Biol. 2021,9, 628157–628184. [CrossRef] 21. Lesnefsky, E.J.; Chen, Q.; Tandler, B.; Hoppel, C.L. Mitochondrial Dysfunction and Myocardial Ischemia-Reperfusion: Implications for Novel Therapies. Annu. Rev. Pharmacol. Toxicol. 2017,57, 535–565. [CrossRef] 22. Paradies, G.; Petrosillo, G.; Paradies, V.; Ruggiero, F.M. Role of Cardiolipin Peroxidation and Ca 2+ in Mitochondrial Dysfunction and Disease. Cell Calcium 2009,45, 643–650. [CrossRef] 23. Dolder, M.; Wendt, S.; Wallimann, T. Mitochondrial Creatine Kinase in Contact Sites: Interaction with Porin and Adenine Nucleotide Translocase, Role in Permeability Transition and Sensitivity to Oxidative Damage. NeuroSignals 2001 ,10, 93–111. [CrossRef] 24. Wagner, G.R.; Bhatt, D.P.; O’Connell, T.M.; Thompson, J.W.; Dubois, L.G.; Backos, D.S.; Yang, H.; Mitchell, G.A.; Ilkayeva, O.R.; Stevens, R.D.; et al. A Class of Reactive Acyl-CoA Species Reveals the Non-Enzymatic Origins of Protein Acylation. Cell Metab. 2017,25, 823–837. [CrossRef] [PubMed] 25. Wagner, G.R.; Hirschey, M.D. Nonenzymatic Protein Acylation as a Carbon Stress Regulated by Sirtuin Deacylases. Mol. Cell 2014,54, 5–16. [CrossRef] [PubMed] 26. Yang, S.J.; Choi, J.M.; Kim, L.; Park, S.E.; Rhee, E.J.; Lee, W.Y.; Oh, K.W.; Park, S.W.; Park, C.Y. Nicotinamide Improves Glucose Metabolism and Affects the Hepatic NAD-Sirtuin Pathway in a Rodent Model of Obesity and Type 2 Diabetes. J. Nutr. Biochem. 2014,25, 66–72. [CrossRef] 27. Tafani, M.; Sansone, L.; Limana, F.; Arcangeli, T.; de Santis, E.; Polese, M.; Fini, M.; Russo, M.A. The Interplay of Reactive Oxygen Species, Hypoxia, Inflammation, and Sirtuins in Cancer Initiation and Progression. Oxidative Med. Cell. Longev. 2016 , 2016, 3907147. [CrossRef] [PubMed] 28. Wai, T.; Langer, T. Mitochondrial Dynamics and Metabolic Regulation. Trends Endocrinol. Metab. 2016 ,27, 105–117. [CrossRef] [PubMed] 29. Mishra, P.; Chan, D.C. Metabolic Regulation of Mitochondrial Dynamics. J. Cell Biol. 2016,212, 379–387. [CrossRef] 30. Palmeira, C.M.; Teodoro, J.S.; Amorim, J.A.; Steegborn, C.; Sinclair, D.A.; Rolo, A.P. Mitohormesis and Metabolic Health_ The Interplay between ROS, CAMP and Sirtuins. Free Radic. Biol. Med. 2019,141, 483–491. [CrossRef] 31. Sastre, J.; Serviddio, G.; Javier, P.; Minana, J.B.; Arduini, A.; Vendemiale, G.; Poli, G.; Pallardo, F.V.; Vina, J. Mitochondrial Function in Liver Disease. Front. Biosci. 2007,12, 1200–1209. [CrossRef] 32. Rasola, A.; Bernardi, P. Mitochondrial Permeability Transition in Ca(2+)-Dependent Apoptosis and Necrosis. Cell Calcium 2011 , 50, 222–233. [CrossRef] 33. Ichas, F.; Mazat, J.P. From Calcium Signaling to Cell Death: Two Conformations for the Mitochondrial Permeability Transition Pore. Switching from Lowto High-Conductance State. Biochim. Biophys. Acta Bioenergy 1998,1366, 33–50. [CrossRef] 34. Suh, D.H.; Kim, M.K.; Kim, H.S.; Chung, H.H.; Song, Y.S. Mitochondrial Permeability Transition Pore as a Selective Target for Anti-Cancer Therapy. Front. Oncol. 2013,3, 41. [CrossRef] 35. Thomas, L.W.; Ashcroft, M. Exploring the Molecular Interface between Hypoxia-Inducible Factor Signalling and Mitochondria. Cell. Mol. Life Sci. 2019,76, 1759–1777. [CrossRef] 36. Marin, W.; Marin, D.; Ao, X.; Liu, Y. Mitochondria as a Therapeutic Target for Cardiac Ischemia-Reperfusion Injury (Review). Int. J. Mol. Med. 2021,47, 485–499. [CrossRef] [PubMed] 37. Li, J.; Li, Y.; Jiao, J.; Wang, J.; Li, Y.; Qin, D.; Li, P. Mitofusin 1 Is Negatively Regulated by MicroRNA 140 in Cardiomyocyte Apoptosis. Mol. Cell. Biol. 2014,34, 1788–1799. [CrossRef] [PubMed] 38. Boengler, K.; Lochnit, G.; Schulz, R. Mitochondria “THE” Target of Myocardial Conditioning. Am. J. Physiol. Heart Circ. Physiol. 2018,315, H1215–H1231. [CrossRef] 39. Go, K.L.; Lee, S.; Zendejas, I.; Behrns, K.E.; Kim, J.S. Mitochondrial Dysfunction and Autophagy in Hepatic Ischemia/Reperfusion Injury. BioMed Res. Int. 2015,2015, 183469. [CrossRef] 40. Hu, C.; Li, L. Pre-Conditions for Eliminating Mitochondrial Dysfunction and Maintaining Liver Function after Hepatic Ischaemia Reperfusion. J. Cell. Mol. Med. 2017,21, 1719–1731. [CrossRef] 41. Cancela, J.M.; Petersen, O.H. Regulation of Intracellular Ca 2+ Stores by Multiple Ca 2+ -Releasing Messengers. Diabetes 2002 ,51 (Suppl. 3), S349–S357. [CrossRef] 42. Raffaello, A.; Mammucari, C.; Gherardi, G.; Rizzuto, R. Calcium at the Center of Cell Signaling: Interplay between Endoplasmic Reticulum, Mitochondria, and Lysosomes. Trends Biochem. Sci. 2016,41, 1035–1049. [CrossRef] 43. Wang, H.G.; Pathan, N.; Ethell, I.M.; Krajewski, S.; Yamaguchi, Y.; Shibasaki, F.; McKeon, F.; Bobo, T.; Franke, T.F.; Reed, J.C. Ca2+-Induced Apoptosis Through Calcineurin Dephosphorylation of BAD. Science 1999,284, 339–343. [CrossRef] [PubMed] Cells 2022,11, 688 18 of 24 44. Ikeda, M.; Ariyoshi, H.; Sakon, M.; Kambayashi, J.I.; Yoshikawa, N.; Shinoki, N.; Kawasaki, T.; Monden, M. A Role for Local Calcium Gradients upon Hypoxic Injury in Human Umbilical Vein Endothelial Cells (HUVEC). Cell Calcium 1998 ,24, 49–57. [CrossRef] 45. Mittal, M.; Siddiqui, M.R.; Tran, K.; Reddy, S.P.; Malik, A.B. Reactive Oxygen Species in Inflammation and Tissue Injury. Antioxid. Redox Signal. 2014,20, 1126–1167. [CrossRef] 46. Zhou, W.; Zhang, Y.; Hosch, M.S.; Lang, A.; Zwacka, R.M.; Engelhardt, J.F. Subcellular Site of Superoxide Dismutase Expression Differentially Controls AP-1 Activity and Injury in Mouse Liver Following Ischemia/Reperfusion. Hepatology 2001 ,33, 902–914. [CrossRef] [PubMed] 47. Nagendra, A.R.; Mickelson, J.K.; Smith, C.W. CD18 Integrin and CD54-Dependent Neutrophil Adhesion to CytokineStimulated Human Hepatocytes. Am. J. Physiol. Gastrointest. Liver Physiol. 1997,272, G408–G416. [CrossRef] 48. Martins, R.M.; Teodoro, J.S.; Furtado, E.; Rolo, A.P.; Palmeira, C.M.; Tralhão, J.G. Recent Insights into Mitochondrial Targeting Strategies in Liver Transplantation. Int. J. Med. Sci. 2018,15, 248–256. [CrossRef] 49. Marchi, S.; Patergnani, S.; Missiroli, S.; Morciano, G.; Rimessi, A.; Wieckowski, M.R.; Giorgi, C.; Pinton, P. Mitochondrial and Endoplasmic Reticulum Calcium Homeostasis and Cell Death. Cell Calcium 2018,69, 62–72. [CrossRef] 50. Duchen, M.R. Mitochondria and Calcium: From Cell Signalling to Cell Death. J. Physiol. 2000,529, 57–68. [CrossRef] 51. Cervantes-Silva, M.P.; Cox, S.L.; Curtis, A.M. Alterations in Mitochondrial Morphology as a Key Driver of Immunity and Host Defence. EMBO Rep. 2021,22, e53086–e53104. [CrossRef] 52. Vringer, E.; Tait, S.W.G. Mitochondria and Inflammation: Cell Death Heats Up. Front. Cell Dev. Biol. 2019,7, 100. [CrossRef] 53. West, A.P.; Shadel, G.S. Mitochondrial DNA in Innate Immune Responses and Inflammatory Pathology. Nat. Rev. Immunol. 2017 , 17, 363–375. [CrossRef] [PubMed] 54. Zhong, F.; Liang, S.; Zhong, Z. Emerging Role of Mitochondrial DNA as a Major Driver of Inflammation and Disease Progression. Trends Immunol. 2019,40, 1120–1133. [CrossRef] [PubMed] 55. Riley, J.S.; Tait, S.W. Mitochondrial DNA in Inflammation and Immunity. EMBO Rep. 2020 ,21, e49799–e49816. [CrossRef] [PubMed] 56. Little, J.P.; Simtchouk, S.; Schindler, S.M.; Villanueva, E.B.; Gill, N.E.; Walker, D.G.; Wolthers, K.R.; Klegeris, A. Mitochondrial Transcription Factor A (Tfam) Is a pro-Inflammatory Extracellular Signaling Molecule Recognized by Brain Microglia. Mol. Cell. Neurosci. 2014,60, 88–96. [CrossRef] [PubMed] 57. Chaung, W.W.; Wu, R.; Ji, Y.; Dong, W.; Wang, P. Mitochondrial Transcription Factor A Is a Proinflammatory Mediator in Hemorrhagic Shock. Int. J. Mol. Med. 2012,30, 199–203. [CrossRef] [PubMed] 58. Collins, L.V.; Hajizadeh, S.; Holme, E.; Jonsson, I.-M.; Tarkowski, A. Endogenously Oxidized Mitochondrial DNA Induces in Vivo and in Vitro Inflammatory Responses. J. Leukoc. Biol. 2004,75, 995–1000. [CrossRef] [PubMed] 59. Zhou, R.; Yazdi, A.S.; Menu, P.; Tschopp, J. A Role for Mitochondria in NLRP3 Inflammasome Activation. Nature 2011 ,469, 221–226. [CrossRef] [PubMed] 60. Liu, Q.; Zhang, D.; Hu, D.; Zhou, X.; Zhou, Y. The Role of Mitochondria in NLRP3 Inflammasome Activation. Mol. Immunol. 2018 , 103, 115–124. [CrossRef] 61. Zhong, Z.; Liang, S.; Sanchez-Lopez, E.; He, F.; Shalapour, S.; Lin, X.J.; Wong, J.; Ding, S.; Seki, E.; Schnabl, B.; et al. New Mitochondrial DNA Synthesis Enables NLRP3 Inflammasome Activation. Nature 2018,560, 198–203. [CrossRef] 62. Shimada, K.; Crother, T.R.; Karlin, J.; Dagvadorj, J.; Chiba, N.; Chen, S.; Ramanujan, V.K.; Wolf, A.J.; Vergnes, L.; Ojcius, D.M.; et al. Oxidized Mitochondrial DNA Activates the NLRP3 Inflammasome during Apoptosis. Immunity 2012 ,36, 401–414. [CrossRef] [PubMed] 63. Lupfer, C.; Thomas, P.G.; Anand, P.K.; Vogel, P.; Milasta, S.; Martinez, J.; Huang, G.; Green, M.; Kundu, M.; Chi, H.; et al. Receptor Interacting Protein Kinase 2-Mediated Mitophagy Regulates Inflammasome Activation during Virus Infection. Nat. Immunol. 2013,14, 480–488. [CrossRef] [PubMed] 64. Hu, Q.; Wood, C.R.; Cimen, S.; Venkatachalam, A.B.; Alwayn, I.P.J. Mitochondrial Damage-Associated Molecular Patterns (MTDs) Are Released during Hepatic Ischemia Reperfusion and Induce Inflammatory Responses. PLoS ONE 2015 ,10, e0140122. [CrossRef] [PubMed] 65. Missiroli, S.; Genovese, I.; Perrone, M.; Vezzani, B.; Vitto, V.A.M.; Giorgi, C. The Role of Mitochondria in Inflammation: From Cancer to Neurodegenerative Disorders. J. Clin. Med. 2020,9, 740. [CrossRef] 66. Selzner, N.; Rudiger, H.; Graf, R.; Clavien, P.A. Protective Strategies against Ischemic Injury of the Liver. Gastroenterology 2003 , 125, 917–936. [CrossRef] 67. Selzner, N.; Selzner, M.; Jochum, W.; Clavien, P.A. Ischemic Preconditioning Protects the Steatotic Mouse Liver against Reperfusion Injury: An ATP Dependent Mechanism. J. Hepatol. 2003,39, 55–61. [CrossRef] 68. Martins, R.M.; Teodoro, J.S.; Furtado, E.; Rolo, A.P.; Palmeira, C.M.; Tralhão, J.G. Evaluation of Bioenergetic and Mitochondrial Function in Liver Transplantation. Clin. Mol. Hepatol. 2019,25, 190–198. [CrossRef] 69. Alexandrino, H.; Varela, A.T.; Teodoro, J.S.; Martins, M.A.; Rolo, A.P.; Tralhão, J.G.; Palmeira, C.M.; Castro e Sousa, F. Mitochondrial Bioenergetics and Posthepatectomy Liver Dysfunction. Eur. J. Clin. Investig. 2016,46, 627–635. [CrossRef] 70. Alexandrino, H.; Rolo, A.; Teodoro, J.S.; Donato, H.; Martins, R.; Serôdio, M.; Martins, M.; Tralhão, J.G.; Caseiro Alves, F.; Palmeira, C.; et al. Bioenergetic Adaptations of the Human Liver in the ALPPS Procedure—How Liver Regeneration Correlates with Mitochondrial Energy Status. HPB 2017,19, 1091–1103. [CrossRef] Cells 2022,11, 688 19 of 24 71. Kong, W.N.; Li, W.; Bai, C.; Dong, Y.; Wu, Y.; An, W. Augmenter of Liver Regeneration-Mediated Mitophagy Protects against Hepatic Ischemia/Reperfusion Injury. Am. J. Transplant. 2021,22, 130–143. [CrossRef] 72. Gu, J.; Zhang, T.; Guo, J.; Chen, K.; Li, H.; Wang, J. PINK1 Activation and Translocation to Mitochondria-Associated Membranes Mediates Mitophagy and Protects Against Hepatic Ischemia/Reperfusion Injury. Shock 2020,54, 783–793. [CrossRef] 73. Sun, K.; Liu, Z.S.; Sun, Q. Role of Mitochondria in Cell Apoptosis during Hepatic Ischema-Reperfusion Injury and Protective Effect of Ischemic Postconditioning. World J. Gastroenterol. 2004,10, 1934–1938. [CrossRef] [PubMed] 74. Zheng, J.; Chen, L.; Lu, T.; Zhang, Y.; Sui, X.; Li, Y.; Huang, X.; He, L.; Cai, J.; Zhou, C.; et al. MSCs Ameliorate Hepatocellular Apoptosis Mediated by PINK1-Dependent Mitophagy in Liver Ischemia/Reperfusion Injury through AMPK α Activation. Cell Death Dis. 2020,11, 256–275. [CrossRef] [PubMed] 75. Liu, A.; Fang, H.; Wei, W.; Dirsch, O.; Dahmen, U. Ischemic Preconditioning Protects against Liver Ischemia/Reperfusion Injury via Heme Oxygenase-1-Mediated Autophagy. Crit. Care Med. 2014,42, e762–e771. [CrossRef] [PubMed] 76. Wang, Y.; Shen, J.; Xiong, X.; Xu, Y.; Zhang, H.; Huang, C.; Tian, Y.; Jiao, C.; Wang, X.; Li, X. Remote Ischemic Preconditioning Protects against Liver Ischemia-Reperfusion Injury via Heme Oxygenase-1-Induced Autophagy. PLoS ONE 2014 ,9, e98846. [CrossRef] 77. Clavien, P.A.; Yadav, S.; Sindram, D.; Bentley, R.C. Protective Effects of Ischemic Preconditioning for Liver Resection Performed under Inflow Occlusion in Humans. Ann. Surg. 2000,232, 155–162. [CrossRef] 78. Rüdiger, H.A.; Graf, R.; Clavien, P.A. Sub-Lethal Oxidative Stress Triggers the Protective Effects of Ischemic Preconditioning in the Mouse Liver. J. Hepatol. 2003,39, 972–977. [CrossRef] 79. Tejima, K.; Arai, M.; Ikeda, H.; Tomiya, T.; Yanase, M.; Inoue, Y.; Nagashima, K.; Nishikawa, T.; Watanabe, N.; Omata, M.; et al. Ischemic Preconditioning Protects Hepatocytes via Reactive Oxygen Species Derived from Kupffer Cells in Rats. Gastroenterology 2004,127, 1488–1496. [CrossRef] 80. Petrowsky, H.; McCormack, L.; Trujillo, M.; Selzner, M.; Jochum, W.; Clavien, P.A. A Prospective, Randomized, Controlled Trial Comparing Intermittent Portal Triad Clamping versus Ischemic Preconditioning with Continuous Clamping for Major Liver Resection. Ann. Surg. 2006,244, 921–928. [CrossRef] 81. Rüdiger, H.A.; Kang, K.J.; Sindram, D.; Riehle, H.M.; Clavien, P.A. Comparison of Ischemic Preconditioning and Intermittent and Continuous Inflow Occlusion in the Murine Liver. Ann. Surg. 2002,235, 400–407. [CrossRef] 82. Ben Mosbah, I.; Duval, H.; Mbatchi, S.F.; Ribault, C.; Grandadam, S.; Pajaud, J.; Morel, F.; Boudjema, K.; Compagnon, P.; Corlu, A. Intermittent Selective Clamping Improves Rat Liver Regeneration by Attenuating Oxidative and Endoplasmic Reticulum Stress. Cell Death Dis. 2014,5, e1107–e1118. [CrossRef] [PubMed] 83. Ricca, L.; Lemoine, A.; Cauchy, F.; Hamelin, J.; Sebagh, M.; Esposti, D.D.; Salloum, C.; Vibert, E.; Balducci, G.; Azoulay, D. Ischemic Postconditioning of the Liver Graft in Adult Liver Transplantation. Transplantation 2015 ,99, 1633–1643. [CrossRef] [PubMed] 84. Monbaliu, D.; Liu, Q.; Libbrecht, L.; de Vos, R.; Vekemans, K.; Debbaut, C.; Detry, O.; Roskams, T.; van Pelt, J.; Pirenne, J. Preserving the Morphology and Evaluating the Quality of Liver Grafts by Hypothermic Machine Perfusion: A Proof-of-Concept Study Using Discarded Human Livers. Liver Transplant. 2012,18, 1495–1507. [CrossRef] [PubMed] 85. Kim, W.H.; Lee, J.-H.; Ko, J.S.; Min, J.J.; Gwak, M.S.; Kim, G.S.; Lee, S.K. Effect of Remote Ischemic Postconditioning on Patients Undergoing Living Donor Liver Transplantation. Liver Transplant. 2014,20, 1383–1392. [CrossRef] 86. Tara, A.; Dominic, J.L.; Patel, J.N.; Garg, I.; Yeon, J.; Memon, M.S.; Gergal Gopalkrishna Rao, S.R.; Bugazia, S.; Dhandapani, T.P.M.; Kannan, A.; et al. Mitochondrial Targeting Therapy Role in Liver Transplant Preservation Lines: Mechanism and Therapeutic Strategies. Cureus 2021,13, e16599. [CrossRef] 87. Tapuria, N.; Junnarkar, S.; Abu-Amara, M.; Fuller, B.; Seifalian, A.M.; Davidson, B.R. Modulation of Microcirculatory Changes in the Late Phase of Hepatic Ischaemia-Reperfusion Injury by Remote Ischaemic Preconditioning. HPB 2012 ,14, 87–97. [CrossRef] 88. Zhou, H.; Li, L.; Sun, H.; Li, H.; Wu, Y.; Zhang, X.; Zhang, J. Remote Ischemic Preconditioning Attenuates Hepatic Ischemia/Reperfusion Injury after Hemorrhagic Shock by Increasing Autophagy. Int. J. Med. Sci. 2021,18, 873–882. [CrossRef] 89. Choi, E.K.; Jung, H.; Jeon, S.; Lim, J.A.; Lee, J.; Kim, H.; Hong, S.W.; Jang, M.H.; Lim, D.G.; Kwak, K.H. Role of Remote Ischemic Preconditioning in Hepatic Ischemic Reperfusion Injury. Dose-Response 2020 ,18, 1559325820946923–1559325820946929. [CrossRef] 90. Kanoria, S.; Robertson, F.P.; Mehta, N.N.; Fusai, G.; Sharma, D.; Davidson, B.R. Effect of Remote Ischaemic Preconditioning on Liver Injury in Patients Undergoing Major Hepatectomy for Colorectal Liver Metastasis: A Pilot Randomised Controlled Feasibility Trial. World J. Surg. 2017,41, 1322–1330. [CrossRef] 91. Ren, Y.; Lin, S.; Liu, W.; Ding, H. Hepatic Remote Ischemic Preconditioning (RIPC) Protects Heart Damages Induced by Ischemia Reperfusion Injury in Mice. Front. Physiol. 2021,12, 713564–713571. [CrossRef] 92. van Rijn, R.; Schurink, I.J.; de Vries, Y.; van den Berg, A.P.; Cortes Cerisuelo, M.; Darwish Murad, S.; Erdmann, J.I.; Gilbo, N.; de Haas, R.J.; Heaton, N.; et al. Hypothermic Machine Perfusion in Liver Transplantation—A Randomized Trial. N. Engl. J. Med. 2021,384, 1391–1401. [CrossRef] [PubMed] 93. Ravikumar, R.; Jassem, W.; Mergental, H.; Heaton, N.; Mirza, D.; Perera, M.T.P.R.; Quaglia, A.; Holroyd, D.; Vogel, T.; Coussios, C.C.; et al. Liver Transplantation After Ex Vivo Normothermic Machine Preservation: A Phase 1 (First-in-Man) Clinical Trial. Am. J. Transplant. 2016,16, 1779–1787. [CrossRef] [PubMed] 94. Lin, H.C.; Liu, S.Y.; Lai, H.S.; Lai, I.R. Isolated Mitochondria Infusion Mitigates Ischemia-Reperfusion Injury of the Liver in Rats. Shock 2013,39, 304–310. [CrossRef] Cells 2022,11, 688 20 of 24 95. Hayashida, K.; Takegawa, R.; Shoaib, M.; Aoki, T.; Choudhary, R.C.; Kuschner, C.E.; Nishikimi, M.; Miyara, S.J.; Rolston, D.M.; Guevara, S.; et al. Mitochondrial Transplantation Therapy for Ischemia Reperfusion Injury: A Systematic Review of Animal and Human Studies. J. Transl. Med. 2021,19, 214–229. [CrossRef] [PubMed] 96. Yamada, Y.; Ito, M.; Arai, M.; Hibino, M.; Tsujioka, T.; Harashima, H. Challenges in Promoting Mitochondrial Transplantation Therapy. Int. J. Mol. Sci. 2020,21, 6365. [CrossRef] 97. Fu, A.; Shi, X.; Zhang, H.; Fu, B. Mitotherapy for Fatty Liver by Intravenous Administration of Exogenous Mitochondria in Male Mice. Front. Pharmacol. 2017,8, 239–241. [CrossRef] 98. Soares, R.O.S.; Losada, D.M.; Jordani, M.C.; Évora, P.; Castro-E-Silva, O. Ischemia/Reperfusion Injury Revisited: An Overview of the Latest Pharmacological Strategies. Int. J. Mol. Sci. 2019,20, 5034. [CrossRef] 99. Xu, C.; Wang, J.; Fan, Z.; Zhang, S.; Qiao, R.; Liu, Y.; Yang, J.; Yang, L.; Wang, H. Cardioprotective Effects of Melatonin against Myocardial Ischaemia/Reperfusion Injury: Activation of AMPK/Nrf2 Pathway. J. Cell. Mol. Med. 2021 ,25, 6455–6459. [CrossRef] 100. Kalimeris, K.; Briassoulis, P.; Ntzouvani, A.; Nomikos, T.; Papaparaskeva, K.; Politi, A.; Batistaki, C.; Kostopanagiotou, G. N-Acetylcysteine Ameliorates Liver Injury in a Rat Model of Intestinal Ischemia Reperfusion. J. Surg. Res. 2016 ,206, 263–272. [CrossRef] 101. Liu, Z.; Chen, J.M.; Huang, H.; Kuznicki, M.; Zheng, S.; Sun, W.; Quan, N.; Wang, L.; Yang, H.; Guo, H.M.; et al. The Protective Effect of Trimetazidine on Myocardial Ischemia/Reperfusion Injury through Activating AMPK and ERK Signaling Pathway. Metab. Clin. Exp. 2016,65, 122–130. [CrossRef] 102. Teodoro, J.S.; Varela, A.T.; Duarte, F.V.; Gomes, A.P.; Palmeira, C.M.; Rolo, A.P. Indirubin and NAD + Prevent Mitochondrial Ischaemia/Reperfusion Damage in Fatty Livers. Eur. J. Clin. Investig. 2018,48, e12932. [CrossRef] [PubMed] 103. Zhang, Y.; Wang, Y.; Xu, J.; Tian, F.; Hu, S.; Chen, Y.; Fu, Z. Melatonin Attenuates Myocardial Ischemia-Reperfusion Injury via Improving Mitochondrial Fusion/Mitophagy and Activating the AMPK-OPA1 Signaling Pathways. J. Pineal Res. 2019 ,66, e12542. [CrossRef] [PubMed] 104. Zhai, M.; Li, B.; Duan, W.; Jing, L.; Zhang, B.; Zhang, M.; Yu, L.; Liu, Z.; Yu, B.; Ren, K.; et al. Melatonin Ameliorates Myocardial Ischemia Reperfusion Injury through SIRT3-Dependent Regulation of Oxidative Stress and Apoptosis. J. Pineal Res. 2017 , 63, e12419. [CrossRef] [PubMed] 105. Chen, H.H.; Chen, Y.T.; Yang, C.C.; Chen, K.H.; Sung, P.H.; Chiang, H.J.; Chen, C.H.; Chua, S.; Chung, S.Y.; Chen, Y.L.; et al. Melatonin Pretreatment Enhances the Therapeutic Effects of Exogenous Mitochondria against Hepatic Ischemia–Reperfusion Injury in Rats through Suppression of Mitochondrial Permeability Transition. J. Pineal Res. 2016,61, 52–68. [CrossRef] 106. Jegatheeswaran, S.; Siriwardena, A.K. Experimental and Clinical Evidence for Modification of Hepatic Ischaemia-Reperfusion Injury by N-Acetylcysteine during Major Liver Surgery. HPB 2011,13, 71–78. [CrossRef] 107. Sun, Y.; Pu, L.Y.; Lu, L.; Wang, X.H.; Zhang, F.; Rao, J.H. N-Acetylcysteine Attenuates Reactive-Oxygen-Speciesmediated Endoplasmic Reticulum Stress during Liver Ischemia-Reperfusion Injury. World J. Gastroenterol. 2014 ,20, 15289–15298. [CrossRef] 108. Cayuela, N.C.; Koike, M.K.; de Fátima Jacysyn, J.; Rasslan, R.; Cerqueira, A.R.A.; Costa, S.K.P.; Diniz-Júnior, J.A.P.; Utiyama, E.M.; de Souza Montero, E.F. N-Acetylcysteine Reduced Ischemia and Reperfusion Damage Associated with Steatohepatitis in Mice. Int. J. Mol. Sci. 2020,21, 4106. [CrossRef] 109. Varela, A.T.; Rolo, A.P.; Palmeira, C.M. Fatty Liver and Ischemia/Reperfusion: Are There Drugs Able to Mitigate Injury? Curr. Med. Chem. 2011,18, 4987–5002. [CrossRef] 110. Tikhaze, A.K.; Lankin, V.Z.; Zharova, E.A.; Kolycheva, S.V. Trimetazidine as Indirect Antioxidant. Bull. Exp. Biol. Med. 2000 ,130, 951–953. [CrossRef] 111. Pantazi, E.; Zaouali, M.A.; Bejaoui, M.; Folch-Puy, E.; Abdennebi, H.B.; Varela, A.T.; Rolo, A.P.; Palmeira, C.M.; Roselló-Catafau, J. Sirtuin 1 in Rat Orthotopic Liver Transplantation: An IGL-1 Preservation Solution Approach. World J. Gastroenterol. 2015 ,21, 1765–1774. [CrossRef] 112. Varela, A.T.; Simões, A.M.; Teodoro, J.S.; Duarte, F.V.; Gomes, A.P.; Palmeira, C.M.; Rolo, A.P. Indirubin-3’-Oxime Prevents Hepatic I/R Damage by Inhibiting GSK-3βand Mitochondrial Permeability Transition. Mitochondrion 2010,10, 456–463. [CrossRef] 113. Hurst, S.; Gonnot, F.; Dia, M.; Crola Da Silva, C.; Gomez, L.; Sheu, S.S. Phosphorylation of Cyclophilin D at Serine 191 Regulates Mitochondrial Permeability Transition Pore Opening and Cell Death after Ischemia-Reperfusion. Cell Death Dis. 2020 ,11, 661–673. [CrossRef] [PubMed] 114. Martin, J.L.; Costa, A.S.H.; Gruszczyk, A.V.; Beach, T.E.; Allen, F.M.; Prag, H.A.; Hinchy, E.C.; Mahbubani, K.; Hamed, M.; Tronci, L.; et al. Succinate Accumulation Drives Ischaemia-Reperfusion Injury during Organ Transplantation. Nat. Metab. 2019 ,1, 966–974. [CrossRef] [PubMed] 115. Xu, A.; Szczepanek, K.; Hu, Y.; Lesnefsky, E.J.; Chen, Q. Cardioprotection by Modulation of Mitochondrial Respiration during Ischemia-Reperfusion: Role of Apoptosis-Inducing Factor. Biochem. Biophys. Res. Commun. 2013,435, 627–633. [CrossRef] 116. Chouchani, E.T.; Methner, C.; Nadtochiy, S.M.; Logan, A.; Pell, V.R.; Ding, S.; James, A.M.; Cochemé, H.M.; Reinhold, J.; Lilley, K.S.; et al. Cardioprotection by S-Nitrosation of a Cysteine Switch on Mitochondrial Complex i. Nat. Med. 2013 ,19, 753–759. [CrossRef] [PubMed] 117. Portakal, O.; Inal-Erden, M. Effects of Pentoxifylline and Coenzyme Q10 in Hepatic Ischemia/Reperfusion Injury. Clin. Biochem. 1999,32, 461–466. [CrossRef] Cells 2022,11, 688 21 of 24 118. Genova, M.L.; Bonacorsi, E.; D’Aurelio, M.; Formiggini, G.; Nardo, B.; Cuccomarino, S.; Turi, P.; Pich, M.M.; Lenaz, G.; Bovina, C. Protective Effect of Exogenous Coenzyme Q in Rats Subjected to Partial Hepatic Ischemia and Reperfusion. BioFactors 1999 ,9, 345–349. [CrossRef] 119. Cherkashina, D.V.; Sosimchik, I.A.; Semenchenko, O.A.; Volina, V.V.; Petrenko, A.Y. Mitochondria-Targeted Plastoquinone Derivative SkQ 1 Decreases Ischemia-Reperfusion Injury during Liver Hypothermic Storage for Transplantation. Biochemistry 2011,76, 1022–1029. [CrossRef] 120. van Golen, R.F.; Reiniers, M.J.; Marsman, G.; Alles, L.K.; van Rooyen, D.M.; Petri, B.; van der Mark, V.A.; van Beek, A.A.; Meijer, B.; Maas, M.A.; et al. The Damage-Associated Molecular Pattern HMGB1 Is Released Early after Clinical Hepatic Ischemia/Reperfusion. Biochim. Biophys. Acta Mol. Basis Dis. 2019,1865, 1192–1200. [CrossRef] 121. Brown, D.A.; Hale, S.L.; Baines, C.P.; Rio, C.L.D.; Hamlin, R.L.; Yueyama, Y.; Kijtawornrat, A.; Yeh, S.T.; Frasier, C.R.; Stewart, L.M.; et al. Reduction of Early Reperfusion Injury with the Mitochondria-Targeting Peptide Bendavia. J. Cardiovasc. Pharmacol. Ther. 2014,19, 121–132. [CrossRef] 122. Kezic, A.; Spasojevic, I.; Lezaic, V.; Bajcetic, M. Mitochondria-Targeted Antioxidants: Future Perspectives in Kidney Ischemia Reperfusion Injury. Oxidative Med. Cell. Longev. 2016,2016, 2950503. [CrossRef] [PubMed] 123. Musleh, W.; Bruce, A.; Malfroy, B.; Baudry, M. Effects of EUK-8, a Synthetic Catalytic Superoxide Scavenger, on Hypoxiaand Acidosis-Induced Damage in Hippocampal Slices. Neuropharmacology 1994,33, 929–934. [CrossRef] 124. Davila, A.; Liu, L.; Chellappa, K.; Redpath, P.; Nakamaru-Ogiso, E.; Paolella, L.M.; Zhang, Z.; Migaud, M.E.; Rabinowitz, J.D.; Baur, J.A. Nicotinamide Adenine Dinucleotide Is Transported into Mammalian Mitochondria. eLife 2018 ,7, e33246. [CrossRef] [PubMed] 125. Yamamoto, T.; Byun, J.; Zhai, P.; Ikeda, Y.; Oka, S.; Sadoshima, J. Nicotinamide Mononucleotide, an Intermediate of NAD+ Synthesis, Protects the Heart from Ischemia and Reperfusion. PLoS ONE 2014,9, e98972. [CrossRef] [PubMed] 126. Toropova, Y.G.; Pechnikova, N.A.; Zelinskaya, I.A.; Zhuravsky, S.G.; Kornyushin, O.V.; Gonchar, A.I.; Ivkin, D.Y.; Leonova, Y.V.; Karev, V.E.; Karabak, I.A. Nicotinamide Riboside Has Protective Effects in a Rat Model of Mesenteric Ischaemia-Reperfusion. Int. J. Exp. Pathol. 2018,99, 304–311. [CrossRef] [PubMed] 127. Kaur, J.; Kaur, T.; Sharma, A.K.; Kaur, J.; Yadav, H.N.; Pathak, D.; Singh, A.P. Fenofibrate Attenuates Ischemia Reperfusion-Induced Acute Kidney Injury and Associated Liver Dysfunction in Rats. Drug Dev. Res. 2021,82, 412–421. [CrossRef] 128. Somi, M.H.; Hajipour, B.; Asl, N.A.; Estakhri, R.; Azar, A.N.; Zade, M.N.; Haghjou, A.G.; Vatankhah, A.M. Pioglitazone Attenuates Ischemia/Reperfusion-Induced Liver Injury in Rats. Transplant. Proc. 2009,41, 4105–4109. [CrossRef] 129. Zhang, M.; Yang, D.; Gong, X.; Ge, P.; Dai, J.; Lin, L.; Zhang, L. Protective Benefits of AMP-Activated Protein Kinase in Hepatic Ischemia-Reperfusion Injury. Am. J. Transl. Res. 2017,9, 823–829. 130. Bi, J.; Zhang, J.; Ren, Y.; Du, Z.; Li, Q.; Wang, Y.; Wei, S.; Yang, L.; Zhang, J.; Liu, C.; et al. Irisin Alleviates Liver IschemiaReperfusion Injury by Inhibiting Excessive Mitochondrial Fission, Promoting Mitochondrial Biogenesis and Decreasing Oxidative Stress. Redox Biol. 2019,20, 296–306. [CrossRef] 131. Dusabimana, T.; Kim, S.R.; Kim, H.J.; Kim, H.; Park, S.W. Nobiletin Ameliorates Hepatic Ischemia and Reperfusion Injury through the Activation of SIRT-1/FOXO3a-Mediated Autophagy and Mitochondrial Biogenesis. Exp. Mol. Med. 2019 ,51, 1–16. [CrossRef] 132. Yu, X.; Jia, L.; Yu, W.; Du, H. Dephosphorylation by Calcineurin Regulates Translocation of Dynamin-Related Protein 1 to Mitochondria in Hepatic Ischemia Reperfusion Induced Hippocampus Injury in Young Mice. Brain Res. 2019 ,1711, 68–76. [CrossRef] 133. de la Cruz López, K.G.; Toledo Guzmán, M.E.; Sánchez, E.O.; García Carrancá, A. MTORC1 as a Regulator of Mitochondrial Functions and a Therapeutic Target in Cancer. Front. Oncol. 2019,9, 1373–1395. [CrossRef] [PubMed] 134. Zhang, T.; Guo, J.; Gu, J.; Chen, K.; Li, H.; Wang, J. Protective Role of MTOR in Liver Ischemia/Reperfusion Injury: Involvement of Inflammation and Autophagy. Oxidative Med. Cell. Longev. 2019,2019, 7861290. [CrossRef] [PubMed] 135. Xu, M.; Hang, H.; Huang, M.; Li, J.; Xu, D.; Jiao, J.; Wang, F.; Wu, H.; Sun, X.; Gu, J.; et al. DJ-1 Deficiency in Hepatocytes Improves Liver Ischemia-Reperfusion Injury by Enhancing Mitophagy. CMGH 2021,12, 567–584. [CrossRef] 136. Shi, Q.; Zhao, G.; Wei, S.; Guo, C.; Wu, X.; Zhao, R.C.; Di, G. Pterostilbene Alleviates Liver Ischemia/Reperfusion Injury via PINK1-Mediated Mitophagy. J. Pharmacol. Sci. 2022,148, 19–30. [CrossRef] [PubMed] 137. Konuko ˇ glu, D.; Ta¸sci, I.; Çetinkale, O. Effects of Cyclosporin A and Ibuprofen on Liver Ischemia-Reperfusion Injury in the Rat. Clin. Chim. Acta 1998,275, 1–8. [CrossRef] 138. Travis, D.L.; Fabia, R.; Netto, G.G.; Husberg, B.S.; Goldstein, R.M.; Klintmalm, G.B.; Levy, M.F. Protection by Cyclosporine a against Normothermic Liver IschemiaReperfusion in Pigs. J. Surg. Res. 1998,75, 116–126. [CrossRef] 139. Theruvath, T.P.; Zhong, Z.; Pediaditakis, P.; Ramshesh, V.K.; Currin, R.T.; Tikunov, A.; Holmuhamedov, E.; Lemasters, J.J. Minocycline and N-Methyl-4-Isoleucine Cyclosporin (NIM811) Mitigate Storage/Reperfusion Injury after Rat Liver Transplantation through Suppression of the Mitochondrial Permeability Transition. Hepatology 2008,47, 236–246. [CrossRef] 140. Clarke, S.J.; McStay, G.P.; Halestrap, A.P. Sanglifehrin A Acts as a Potent Inhibitor of the Mitochondrial Permeability Transition and Reperfusion Injury of the Heart by Binding to Cyclophilin-D at a Different Site from Cyclosporin A. J. Biol. Chem. 2002 ,277, 34793–34799. [CrossRef] 141. Chen, C.H.; Ferreira, J.C.B.; Gross, E.R.; Mochly-Rosen, D. Targeting Aldehyde Dehydrogenase 2: New Therapeutic Opportunities. Physiol. Rev. 2014,94, 1–34. [CrossRef] Cells 2022,11, 688 22 of 24 142. Zaouali, M.A.; Abdennebi, H.B.; Padrissa-Altés, S.; Alfany-Fernandez, I.; Rimola, A.; Roselló-Catafau, J. How Institut Georges Lopez Preservation Solution Protects Nonsteatotic and Steatotic Livers against Ischemia-Reperfusion Injury. Transplant. Proc. 2011,43, 77–79. [CrossRef] 143. Ding, J.; Zhang, Q.; Luo, Q.; Ying, Y.; Liu, Y.; Li, Y.; Wei, W.; Yan, F.; Zhang, H. Alda-1 Attenuates Lung Ischemia-Reperfusion Injury by Reducing 4-Hydroxy-2-Nonenal in Alveolar Epithelial Cells. Crit. Care Med. 2016,44, e544. [CrossRef] [PubMed] 144. Zhu, Q.; He, G.; Wang, J.; Wang, Y.; Chen, W. Pretreatment with the ALDH2 Agonist Alda-1 Reduces Intestinal Injury Induced by Ischaemia and Reperfusion in Mice. Clin. Sci. 2017,131, 1123–1136. [CrossRef] [PubMed] 145. Li, M.; Xu, M.; Li, J.; Chen, L.; Xu, D.; Tong, Y.; Zhang, J.; Wu, H.; Kong, X.; Xia, Q. Alda-1 Ameliorates Liver Ischemia-Reperfusion Injury by Activating Aldehyde Dehydrogenase 2 and Enhancing Autophagy in Mice. J. Immunol. Res. 2018 ,2018, 9807139. [CrossRef] 146. Liu, Z.; Ye, S.; Zhong, X.; Wang, W.; Lai, C.H.; Yang, W.; Yue, P.; Luo, J.; Huang, X.; Zhong, Z.; et al. Pretreatment with the ALDH2 Activator Alda-1 Protects Rat Livers from Ischemia/Reperfusion Injury by Inducing Autophagy. Mol. Med. Rep. 2020 ,22, 2373–2385. [CrossRef] [PubMed] 147. Puts, C.F.; Berendsen, T.A.; Bruinsma, B.G.; Ozer, S.; Luitje, M.; Usta, O.B.; Yarmush, M.L.; Uygun, K. Polyethylene Glycol Protects Primary Hepatocytes during Supercooling Preservation. Cryobiology 2015,71, 125–129. [CrossRef] 148. Zaouali, M.A.; Bejaoui, M.; Calvo, M.; Folch-Puy, E.; Pantazi, E.; Pasut, G.; Rimola, A.; Abdennebi, H.B.; Adam, R.; RosellóCatafau, J. Polyethylene Glycol Rinse Solution: An Effective Way to Prevent Ischemia-Reperfusion Injury. World J. Gastroenterol. 2014,20, 16203–16214. [CrossRef] 149. Ferrero-Andrés, A.; Panisello-Roselló, A.; Serafín, A.; Roselló-Catafau, J.; Folch-Puy, E. Polyethylene Glycol 35 (PEG35) Protects against Inflammation in Experimental Acute Necrotizing Pancreatitis and Associated Lung Injury. Int. J. Mol. Sci. 2020 ,21, 917. [CrossRef] 150. Ackland, G.L.; Gutierrez Del Arroyo, A.; Yao, S.T.; Stephens, R.C.; Dyson, A.; Klein, N.J.; Singer, M.; Gourine, A.V. Low-MolecularWeight Polyethylene Glycol Improves Survival in Experimental Sepsis. Crit. Care Med. 2010,38, 629–636. [CrossRef] 151. Panisello-Roselló, A.; Lopez, A.; Folch-Puy, E.; Carbonell, T.; Rolo, A.; Palmeira, C.; Adam, R.; Net, M.; Roselló-Catafau, J. Role of Aldehyde Dehydrogenase 2 in Ischemia Reperfusion Injury: An Update. World J. Gastroenterol. 2018,24, 2984–2994. [CrossRef] 152. Panisello-Roselló, A.; Alva, N.; Flores, M.; Lopez, A.; Benítez, C.C.; Folch-Puy, E.; Rolo, A.; Palmeira, C.; Adam, R.; Carbonell, T.; et al. Aldehyde Dehydrogenase 2 (ALDH2) in Rat Fatty Liver Cold Ischemia Injury. Int. J. Mol. Sci. 2018 ,19, 2479. [CrossRef] [PubMed] 153. Bardallo, R.G.; da Silva, R.T.; Carbonell, T.; Folch-Puy, E.; Palmeira, C.; Roselló-Catafau, J.; Pirenne, J.; Adam, R.; Panisello-Roselló, A. Role of Peg35, Mitochondrial Aldh2, and Glutathione in Cold Fatty Liver Graft Preservation: An Igl-2 Approach. Int. J. Mol. Sci. 2021,22, 5332. [CrossRef] 154. Nehrt, A.; Hamann, K.; Ouyang, H.; Shi, R. Polyethylene Glycol Enhances Axolemmal Resealing Following Transection in Cultured Cells and in Ex Vivo Spinal Cord. J. Neurotrauma 2010,27, 151–161. [CrossRef] [PubMed] 155. Shi, R. Polyethylene Glycol Repairs Membrane Damage and Enhances Functional Recovery: A Tissue Engineering Approach to Spinal Cord Injury. Neurosci. Bull. 2013,29, 460–466. [CrossRef] [PubMed] 156. Xu, X.; Philip, J.L.; Razzaque, M.A.; Lloyd, J.W.; Muller, C.M.; Akhter, S.A. High-Molecular-Weight Polyethylene Glycol Inhibits Myocardial Ischemia-Reperfusion Injury in Vivo. J. Thorac. Cardiovasc. Surg. 2015,149, 588–593. [CrossRef] [PubMed] 157. Bejaoui, M.; Pantazi, E.; Folch-Puy, E.; Panisello, A.; Calvo, M.; Pasut, G.; Rimola, A.; Navasa, M.; Adam, R.; Roselló-Catafau, J. Protective Effect of Intravenous High Molecular Weight Polyethylene Glycol on Fatty Liver Preservation. BioMed Res. Int. 2015 , 2015, 79428. [CrossRef] [PubMed] 158. Bejaoui, M.; Pantazi, E.; Calvo, M.; Folch-Puy, E.; Serafín, A.; Pasut, G.; Panisello, A.; Adam, R.; Roselló-Catafau, J. Polyethylene Glycol Preconditioning: An Effective Strategy to Prevent Liver Ischemia Reperfusion Injury. Oxidative Med. Cell. Longev. 2016 , 2016, 9096549. [CrossRef] 159. Rosello, A.P.; da Silva, R.T.; Castro, C.; Bardallo, R.G.; Calvo, M.; Folch-Puy, E.; Carbonell, T.; Palmeira, C.; Catafau, J.R.; Adam, R. Polyethylene Glycol 35 as a Perfusate Additive for Mitochondrial and Glycocalyx Protection in Hope Liver Preservation. Int. J. Mol. Sci. 2020,21, 5703. [CrossRef] 160. Kahn, J.; Schemmer, P. Control of Ischemia-Reperfusion Injury in Liver Transplantation: Potentials for Increasing the Donor Pool. Visc. Med. 2018,34, 444–448. [CrossRef] 161. Feng, S.; Lai, J.C. Expanded Criteria Donors. Clin. Liver Dis. 2014,18, 633–649. [CrossRef] 162. Peralta, C.; Bulbena, O.; Xaus, C.; Prats, N.; Cutrin, J.C.; Poli, G.; Gelpi, E.; Roselló-Catafau, J. Ischemic Preconditioning: A Defense Mechanism against the Reactive Oxygen Species Generated after Hepatic Ischemia Reperfusion. Transplantation 2002 ,73, 1203–1211. [CrossRef] [PubMed] 163. Robinson, J.R. Control of Water Content of Respiring Kidney Slices by Sodium Chloride and Polyethylene Glycol. J. Physiol. 1978 , 282, 285–294. [CrossRef] [PubMed] 164. Gores, G.J.; Nieminen, A.L.; Wray, B.E.; Herman, B.; Lemasters, J.J. Intracellular PH during “chemical Hypoxia” in Cultured Rat Hepatocytes. Protection by Intracellular Acidosis against the Onset of Cell Death. J. Clin. Investig. 1989 ,83, 386–396. [CrossRef] [PubMed] 165. Jamieson, N.V.; Lindell, S.; Sundberg, R.; Southard, J.H.; Belzer, F.O. An Analysis of the Components in Uw Solution Using the Isolated Perfused Rabbit Liver. Transplantation 1988,46, 512–516. [CrossRef] [PubMed] Cells 2022,11, 688 23 of 24 166. Lee, C.Y.; Mangino, M.J. Preservation Methods for Kidney and Liver. Organogenesis 2009,5, 105–112. [CrossRef] 167. Wang, W.; Hu, X.; Xia, Z.; Liu, Z.; Zhong, Z.; Lu, Z.; Liu, A.; Ye, S.; Cao, Q.; Wang, Y.; et al. Mild Hypothermia Attenuates Hepatic Ischemia-Reperfusion Injury through Regulating the JAK2/STAT3-CPT1a-Dependent Fatty Acid β -Oxidation. Oxidative Med. Cell. Longev. 2020,2020, 5849794. [CrossRef] 168. Czigany, Z.; Lurje, I.; Schmelzle, M.; Schöning, W.; Öllinger, R.; Raschzok, N.; Sauer, I.M.; Tacke, F.; Strnad, P.; Trautwein, C.; et al. Ischemia-Reperfusion Injury in Marginal Liver Grafts and the Role of Hypothermic Machine Perfusion: Molecular Mechanisms and Clinical Implications. J. Clin. Med. 2020,9, 846. [CrossRef] 169. Brüggenwirth, I.M.A.; van Leeuwen, O.B.; de Vries, Y.; Bodewes, S.B.; Adelmeijer, J.; Wiersema-Buist, J.; Lisman, T.; Martins, P.N.; de Meijer, V.E.; Porte, R.J. Extended Hypothermic Oxygenated Machine Perfusion Enables Ex Situ Preservation of Porcine Livers for up to 24 Hours. JHEP Rep. 2020,2, 100092. [CrossRef] 170. Taylor, M.J.; Baicu, S.C. Current State of Hypothermic Machine Perfusion Preservation of Organs: The Clinical Perspective. Cryobiology 2010,60, S20–S35. [CrossRef] 171. Dutkowski, P.; Guarrera, J.V.; de Jonge, J.; Martins, P.N.; Porte, R.J.; Clavien, P.A. Evolving Trends in Machine Perfusion for Liver Transplantation. Gastroenterology 2019,156, 1542–1547. [CrossRef] 172. Collins, G.M.; Bravo-Shugarman, M.; Terasaki, P.I. Kidney Preservation for Transportation. Initial Perfusion and 30 Hours’ Ice Storage. Lancet 1969,2, 1219–1222. [CrossRef] 173. Giraud, S.; Thuillier, R.; Codas, R.; Manguy, E.; Barrou, B.; Valagier, A.; Puichaud, A.; Badet, L.; Nicolas, E.; Eugene, M.; et al. The Optimal Peg for Kidney Preservation: A Preclinical Porcine Study. Int. J. Mol. Sci. 2018,19, 454. [CrossRef] [PubMed] 174. Salahudeen, A.K. Cold Ischemic Injury of Transplanted Kidneys: New Insights from Experimental Studies. Am. J. Physiol. Ren. Physiol. 2004,287, F181–F187. [CrossRef] [PubMed] 175. Hüter, L.; Simon, T.P.; Weinmann, L.; Schuerholz, T.; Reinhart, K.; Wolf, G.; Amann, K.U.; Marx, G. Hydroxyethylstarch Impairs Renal Function and Induces Interstitial Proliferation, Macrophage Infiltration and Tubular Damage in an Isolated Renal Perfusion Model. Crit. Care 2009,13, R23. [CrossRef] [PubMed] 176. Wicomb, W.N.; Hill, J.D.; Avery, J.; Collins, G.M. Optimal Cardioplegia and 24-Hour Heart Storage with Simplified UW Solution Containing Polyethylene Glycol. Transplantation 1990,49, 261–264. [CrossRef] 177. Daniel, M.R.; Wakerley, C.L. Factors Influencing the Survival of Cell Monolayers during Storage at 4 ◦ .Br. J. Exp. Pathol. 1976 ,57, 95–118. 178. Ganote, C.E.; Worstell, J.; Iannotti, J.P.; Kaltenbach, J.P. Cellular Swelling and Irreversible Myocardial Injury. Effects of Polyethylene Glycol and Mannitol in Perfused Rat Hearts. Am. J. Pathol. 1977,88, 95–118. 179. Adam, R.; Delvart, V.; Karam, V.; Ducerf, C.; Navarro, F.; Letoublon, C.; Belghiti, J.; Pezet, D.; Castaing, D.; le Treut, Y.P.; et al. Compared Efficacy of Preservation Solutions in Liver Transplantation: A Long-Term Graft Outcome Study from the European Liver Transplant Registry. Am. J. Transplant. 2015,15, 395–406. [CrossRef] 180. Franco-Gou, R.; Mosbah, I.B.; Serafin, A.; Abdennebi, H.B.; Roselló-Catafau, J.; Peralta, C. New Preservation Strategies for Preventing Liver Grafts against Cold Ischemia Reperfusion Injury. J. Gastroenterol. Hepatol. 2007,22, 1120–1126. [CrossRef] 181. Codas, R.; Petruzzo, P.; Morelon, E.; Lefrançois, N.; Danjou, F.; Berthillot, C.; Contu, P.; Espa, M.; Martin, X.; Badet, L. IGL-1 Solution in Kidney Transplantation: First Multi-Center Study. Clin. Transplant. 2009,23, 337–342. [CrossRef] 182. Dondéro, F.; Paugam-Burtz, C.; Danjou, F.; Stocco, J.; Durand, F.; Belghiti, J. A Randomized Study Comparing IGL-1 to the University of Wisconsin Preservation Solution in Liver Transplantation. Ann. Transplant. 2010,15, 1–7. [PubMed] 183. Hauet, T.; Eugene, M. A New Approach in Organ Preservation: Potential Role of New Polymers. Kidney Int. 2008 ,74, 998–1003. [CrossRef] [PubMed] 184. Valuckaite, V.; Seal, J.; Zaborina, O.; Tretiakova, M.; Testa, G.; Alverdy, J.C. High Molecular Weight Polyethylene Glycol (PEG 15-20) Maintains Mucosal Microbial Barrier Function during Intestinal Graft Preservation. J. Surg. Res. 2013 ,183, 869–875. [CrossRef] [PubMed] 185. Zaouali, M.A.; Mosbah, I.B.; Boncompagni, E.; Abdennebi, H.B.; Mitjavila, M.T.; Bartrons, R.; Freitas, I.; Rimola, A.; RosellóCatafau, J. Hypoxia Inducible Factor-1 α Accumulation in Steatotic Liver Preservation: Role of Nitric Oxide. World J. Gastroenterol. 2010,16, 3499–3509. [CrossRef] [PubMed] 186. Schlegel, A.; Muller, X.; Mueller, M.; Stepanova, A.; Kron, P.; de Rougemont, O.; Muiesan, P.; Clavien, P.A.; Galkin, A.; Meierhofer, D.; et al. Hypothermic Oxygenated Perfusion Protects from Mitochondrial Injury before Liver Transplantation. EBioMedicine 2020,60, 103014–103028. [CrossRef] 187. Czigany, Z.; Lurje, I.; Tolba, R.H.; Neumann, U.P.; Tacke, F.; Lurje, G. Machine Perfusion for Liver Transplantation in the Era of Marginal Organs—New Kids on the Block. Liver Int. 2019,39, 228–249. [CrossRef] 188. Schlegel, A.; Kron, P.; Graf, R.; Dutkowski, P.; Clavien, P.A. Warm vs. Cold Perfusion Techniques to Rescue Rodent Liver Grafts. J. Hepatol. 2014,61, 1267–1275. [CrossRef] 189. Stegemann, J.; Minor, T. Energy Charge Restoration, Mitochondrial Protection and Reversal of Preservation Induced Liver Injury by Hypothermic Oxygenation Prior to Reperfusion. Cryobiology 2009,58, 331–336. [CrossRef] 190. Schlegel, A.; Muller, X.; Dutkowski, P. Hypothermic Machine Preservation of the Liver: State of the Art. Curr. Transplant. Rep. 2018,5, 93–102. [CrossRef] Cells 2022,11, 688 24 of 24 191. Chouchani, E.T.; Pell, V.R.; Gaude, E.; Aksentijevi´c, D.; Sundier, S.Y.; Robb, E.L.; Logan, A.; Nadtochiy, S.M.; Ord, E.N.J.; Smith, A.C.; et al. Ischaemic Accumulation of Succinate Controls Reperfusion Injury through Mitochondrial ROS. Nature 2014 ,515, 431–435. [CrossRef] 192. Kron, P.; Schlegel, A.; Mancina, L.; Clavien, P.A.; Dutkowski, P. Hypothermic Oxygenated Perfusion (HOPE) for Fatty Liver Grafts in Rats and Humans. J. Hepatol. 2018,68, 82–91. [CrossRef] [PubMed] 193. Dutkowski, P.; Furrer, K.; Tian, Y.; Graf, R.; Clavien, P.A. Novel Short-Term Hypothermic Oxygenated Perfusion (HOPE) System Prevents Injury in Rat Liver Graft from Non-Heart Beating Donor. Ann. Surg. 2006,244, 968–976. [CrossRef] 194. Zeng, Y.; Zhang, X.F.; Fu, B.M.; Tarbell, J.M. The Role of Endothelial Surface Glycocalyx in Mechanosensing and Transduction. Adv. Exp. Med. Biol. 2018,1097, 1–27. [PubMed] 195. Schiefer, J.; Faybik, P.; Koch, S.; Tudor, B.; Kollmann, D.; Kuessel, L.; Krenn, C.G.; Berlakovich, G.; Baron, D.M.; Baron-Stefaniak, J. Glycocalyx Damage within Human Liver Grafts Correlates with Graft Injury and Postoperative Graft Function after Orthotopic Liver Transplantation. Transplantation 2019,104, 72–78. [CrossRef] 196. Lopez, A.; Panisello-Rosello, A.; Castro-Benitez, C.; Adam, R. Glycocalyx Preservation and NO Production in Fatty Livers—The Protective Role of High Molecular Polyethylene Glycol in Cold Ischemia Injury. Int. J. Mol. Sci. 2018,19, 2375. [CrossRef]