Ci a ion: A ellano-Ga cía, L.;
Po illo, M.P.; Ma ínez, J.A.;
Mil on-Laskiba , I. Use ulness o
P obio ics in he Managemen o
NAFLD: E idence and In ol ed
Mechanisms o Ac ion om
P eclinical and Human Models. In . J.
Mol. Sci. 2022,23, 3167. h ps://
doi.o g/10.3390/ijms23063167
Academic Edi o : Gio anni Ta an ino
Recei ed: 8 Feb ua y 2022
Accep ed: 12 Ma ch 2022
Published: 15 Ma ch 2022
Publishe ’s No e: MDPI s ays neu al
wi h ega d o ju isdic ional claims in
published maps and ins i u ional a il-
ia ions.
Copy igh : © 2022 by he au ho s.
Licensee MDPI, Basel, Swi ze land.
This a icle is an open access a icle
dis ibu ed unde he e ms and
condi ions o he C ea i e Commons
A ibu ion (CC BY) license (h ps://
c ea i ecommons.o g/licenses/by/
4.0/).
In e na ional Jou nal o
Molecula Sciences
Re iew
Use ulness o P obio ics in he Managemen o NAFLD:
E idence and In ol ed Mechanisms o Ac ion om P eclinical
and Human Models
Lau a A ellano-Ga cía1, Ma ía P. Po illo 1,2,3,* , J. Al edo Ma ínez 2,4 and Iñaki Mil on-Laskiba 2,4
1Nu i ion and Obesi y G oup, Depa men o Pha macy and Food Sciences, Facul y o Pha macy and Lucio
Lasca ay Resea ch Cen e , Uni e si y o he Basque Coun y (UPV/EHU), 01006 Vi o ia-Gas eiz, Spain;
lau ai.a ellano.ga [email p o ec ed]
2CIBER Fisiopa ología de la Obesidad y Nu ición (CIBEROBN), Ins i u o de Salud Ca los III (ISCIII),
28222 Mad id, Spain; [email p o ec ed]g (J.A.M.); [email p o ec ed]g (I.M.-L.)
3BIOARABA Ins i u e o Heal h, 01006 Vi o ia-Gas eiz, Spain
4P ecision Nu i ion and Ca diome abolic Heal h, IMDEA-Food Ins i u e (Mad id Ins i u e o
Ad anced S udies), Campus o In e na ional Excellence (CEI) UAM+CSIC, Spanish Na ional Resea ch
Council, 28049 Mad id, Spain
*Co espondence: ma iapuy[email p o ec ed]; Tel.: +34-945-013-067; Fax: +34-945-013-014
Abs ac :
The p esen e iew aims a analyzing he cu en e idence ega ding p obio ic admin-
is a ion o non-alcoholic a y li e disease (NAFLD) managemen . Addi ionally, he in ol ed
mechanisms o ac ion modula ed by p obio ic adminis a ion, as well as he e en ual limi a ions
o his he apeu ic app oach and po en ial al e na i es, a e discussed. P eclinical s udies ha e
demons a ed ha he adminis a ion o single-s ain p obio ics and p obio ic mix u es e ec i ely
p e en s die -induced NAFLD. In bo h cases, he magni ude o he desc ibed e ec s, as well as he
in ol ed mechanisms o ac ion, a e compa able, including educed li e lipid accumula ion (due
o lipogenesis down egula ion and a y acid oxida ion up egula ion), eco e y o gu mic obio a
composi ion and enhanced in es inal in eg i y. Simila esul s ha e also been epo ed in clinical
ials, whe e he adminis a ion o p obio ics p o ed o be e ec i e in he ea men o NAFLD in
pa ien s ea u ing his li e condi ion. In his case, in o ma ion ega ding he mechanisms o ac ion
unde lying p obio ics-media ed hepa op o ec i e e ec s is sca ce (mainly due o he di icul y o
li e sample collec ion). Since p obio ics adminis a ion ep esen s an inc eased isk o in ec ion in
ulne able subjec s, much a en ion has been paid o pa abio ics and pos bio ics, which seem o be
e ec i e in he managemen o se e al me abolic diseases, and hus ep esen a sui able al e na i e o
p obio ic usage.
Keywo ds: p obio ics; mic obio a; li e s ea osis; NAFLD; in lamma ion; pa abio ics; pos bio ics
1. In oduc ion
The p e alence o ch onic me abolic diseases has been on he ise in he las decades,
becoming a majo heal h p oblem wo ldwide. Despi e he amoun o a en ion ha has
been paid o obesi y, millions o dea hs (up o 2 million by he yea 2010) ha e also been
a ibu ed o li e diseases such as ci hosis and hepa ocellula ca cinoma [
1
]. In his
line, non-alcoholic a y li e disease (NAFLD), also known as me abolic (dys unc ion)-
associa ed a y li e disease (MAFLD), has become he mos p e alen hepa ic al e a ion
in he las yea s [
2
]. Indeed, i is es ima ed ha he p e alence o NAFLD is 20–30%
in adul s, and ha his p e alence may well be highe in indus ialized coun ies [
3
].
This hepa ic condi ion includes ela i ely benign and e e sible s ea osis, cha ac e ized
by excessi e iglyce ide (TG) accumula ion in he li e , along wi h he mo e ha m ul
s age known as non-alcoholic s ea ohepa i is (NASH), ha can p og ess o ci hosis o
e en hepa ocellula ca cinoma [
4
]. In his ega d, besides he a o emen ioned excessi e
In . J. Mol. Sci. 2022,23, 3167. h ps://doi.o g/10.3390/ijms23063167 h ps://www.mdpi.com/jou nal/ijms
In . J. Mol. Sci. 2022,23, 3167 2 o 25
hepa ic lipid accumula ion leading o simple s ea osis, u he e en s such as in lamma ion,
oxida i e s ess and ib osis a e also in ol ed in he p og ession o he disease [5,6].
Due o he a ay o p ocesses ha ha e been iden i ied o pa icipa e in NAFLD de el-
opmen , he once widely assumed “ wo-hi heo y” has been eplaced by he “mul iple-hi
heo y” [
6
]. Acco ding o he la e , whi e adipose issue insulin esis ance plays a majo
ole, impai ing lipolysis and igge ing in lamma ion. All hese impai men s esul in
a g ea e elease o ee a y acids o he blood s eam, which end up in he li e , hus
con ibu ing o excessi e hepa ic lipid accumula ion. Addi ionally, his inc eased hepa ic
a y acid deposi ion also esul s in lipo oxici y and subsequen mi ochond ial dys unc ion,
which in u n inc eases eac i e oxygen species (ROS) p oduc ion and oxida i e s ess, and
ac i a es endoplasmic e iculum s ess [
6
]. Mo eo e , gu mic obio a al e a ions ha e also
been desc ibed as con ibu ing ac o s o NAFLD de elopmen . Impai ed gu mic obio a
composi ion esul s in a g ea e p oduc ion o p o-in lamma o y cy okines, such as umo
nec osis ac o
α
(TNF
α
) and in e leukin 6 (IL-6), as well as mic obial p oduc s wi h
p o-in lamma o y p ope ies, including lipopolysaccha ides (LPS) and unme hyla ed CpG
DNA [
7
]. Mo eo e , inc eased in es inal pe meabili y, esul ing om al e ed igh junc ions,
leads o g ea e ansloca ion o hese p o-in lamma o y media o s in o ci cula ion, which
once eaching he li e , igge he ac i a ion o p o-in lamma o y pa hways in he o gan,
hus con ibu ing o he p og ession o NAFLD in NASH (Figu e 1) [
8
]. As a as he causes
leading o NAFLD de elopmen a e conce ned, excessi e die a y a and/o suga in ake
(specially uc ose) a e conside ed among he main con ibu o s [
9
,
10
]. Indeed, his kind o
die a y pa e n no only p omo es excessi e hepa ic lipid accumula ion (due o enhanced
de no o lipogenesis and impai ed mi ochond ial a y-acid oxida ion), bu i also induces
li e in lamma ion, oxida i e s ess and mi ochond ial dys unc ion, all o which leads o he
p og ession o hepa ic damage [
9
,
10
]. Ne e heless, o he ac o s such as ood p ocessing
o polyphenol con en can also ha e a ole.
Figu e 1.
Simpli ied schema ic ep esen a ion o he e en s conside ed in he mul iple-hi heo y
leading o NAFLD de elopmen . F/B:Fi micu es/Bac e oide es a io; FA: a y acid; FFA: ee a y acid;
HSL: ho mone-sensi i e lipase; IR: insulin esis ance; ROS: eac i e oxygen species; TG: iglyce ide;
VLDL: e y-low-densi y lipop o ein; WAT: whi e adipose issue. ↑: inc ease; ↓: dec ease.
The high p e alence o NAFLD, as well as i s po en ial implica ions in heal h, high-
ligh s he necessi y o e ec i e app oaches in he p e en ion and ea men o his li e
condi ion. Howe e , since no speci ic ea men has been designed so a , con en ional
in e en ions based on die a y ea men and enhanced physical ac i i y leading o body
weigh educ ion a e s ill widely p esc ibed [
11
,
12
]. One o he main easons o using
such an app oach o NAFLD managemen elies on he highe p e alence o his hepa ic
condi ion in obese subjec s. Indeed, acco ding o ecen da a, i is es ima ed, ha NAFLD
is p esen in up o 50–90% o subjec s ea u ing obesi y [
13
]. No wi hs anding ha he
e ec i eness o hese app oaches has been demons a ed, a common low adhe ence e-
qui es u he he apeu ic ools ha may be p esc ibed as complemen a y o al e na i e
ea men s. In his scena io, he adminis a ion o p obio ics o NAFLD has gained much
In . J. Mol. Sci. 2022,23, 3167 3 o 25
a en ion, especially due o he in ol emen o gu mic obio a al e a ions in he de elop-
men o his li e al e a ion. By de ini ion, p obio ics a e iable mic oo ganisms ha exe
heal h bene i s when consumed in su icien amoun s [
14
]. Thus, p obio ic consump ion
may help no malize gu mic obio a composi ion in pa ien s wi h NAFLD, which in u n
could esul in imp o ed gu ba ie unc ion and dec eased p o-in lamma o y cy okine
p oduc ion and elease. Addi ionally, he eco e y o gu mic obio a eubiosis will also be
help ul in es o ing he p oduc ion and le els o gu mic obio a-de i ed me aboli es wi h
desc ibed heal h bene i s, such as sho -chain a y acids (SCFA) [15].
In his con ex , he aim o his na a i e e iew is o summa ize he a ailable e idence
ega ding p obio ic use ulness in NAFLD p e en ion. In addi ion, he mechanisms o
ac ion desc ibed so a unde lying he po en ial hepa op o ec i e e ec s o p obio ics a e
also discussed. Fo his pu pose, he i s pa o his manusc ip is ocused on he esul s
ob ained in p eclinical s udies ( oden models), whe eas he second pa summa izes he
cu en e idence ob ained om clinical ials. Addi ionally, limi a ions ela ed o p obio ic
in ake, as well as po en ial al e na i es, a e also discussed. Wi h ega d o he c i e ia
ollowed o include o exclude a icles in his na a i e e iew, hose using p obio ics
alone (single s ain o mix u es) o NAFLD (no NASH) managemen , and s udying
a iables such as li e a con en , li e his ologic analysis (li e lipid con en and/o
in lamma ion) and ansaminase le els, we e selec ed. In he con a y, a icles whe e
none o hese a iables we e analyzed o p obio ics we e adminis e ed along wi h o he
ing edien s (unsa u a ed a y acids o polysaccha ides, o ins ance) we e excluded. This
a icle-selec ion ask was ca ied ou by wo di e en pe sons.
2. E ec s o P obio ic Adminis a ion (Single S ain and Mix u es) on NAFLD
P e en ion: E idence om P eclinical S udies
When analyzing he po en ial use ulness o a molecule/compound in NAFLD p e-
en ion, bo h he molecule/compound and he s esso leading o he de elopmen o his
hepa ic condi ion a e adminis e ed oge he . In p eclinical s udies, his li e al e a ion is
commonly induced by using unbalanced die s cha ac e ized by a high con en o a and/o
p ocessed suga . These eeding condi ions no only esul in an impai ed nu ien in ake,
bu hey can also lead o an excessi e calo ic consump ion. Mo eo e , die s lacking speci ic
nu ien s, such as choline-de icien die s, a e also an e ec i e app oach when gene a ing
die -induced NAFLD.
2.1. P eclinical S udies Using Single S ain P obio ics
Di e en s udies ha e been ca ied ou using a single-s ain p obio ic (Table 1). In
gene al, he majo i y o hese s udies ha e add essed he e ec s o speci ic p obio ic s ains
in animals challenged by die s leading o NAFLD. In his line, he adminis a ion o se e al
p obio ic s ains (mainly Lac obacillus and Bi idobac e ium) has been shown o be e ec-
i e in educing li e lipid accumula ion unde die a y condi ions p o iding
40 o 65%
o ene gy as a [
16
–
31
]. Mo eo e , his e ec is also main ained when excessi e hep-
a ic lipid accumula ion is media ed by die a y condi ions p o iding high suga in akes
(1
0 o 30%
o ene gy as uc ose). In his case, p obio ic adminis a ion p e en ed li e lipid
accumula ion when s anda d die s we e supplemen ed wi h uc ose [
32
–
35
], as well as
when high- a and uc ose in akes occu ed concomi an ly [
33
,
34
]. Fu he mo e, p obio ic-
adminis a ion-media ed li e a accumula ion p e en ion was also epo ed in a s udy in
which NAFLD was induced using a choline-de icien die [
36
]. I is wo h no ing ha he
a o emen ioned e ec s we e desc ibed in bo h mice and a s ecei ing di e en p obio ic
doses ( om
1×107 o 1 ×1010 CFU/day
) and du ing di e en ea men pe iods ( om
4 o 42 weeks) (Table 1).
In . J. Mol. Sci. 2022,23, 3167 4 o 25
Table 1. P eclinical s udies ( oden models) add essing he e ec s o di e en single-s ain p obio ics on die -induced NAFLD.
Re e ence Animal Model Expe imen al Condi ions P obio ic T ea men E ec s on Li e Mechanisms o Ac ion
[32]Female C57BL/J6 mice
6-week-old
STD die wi h 30% uc ose
in d inking wa e .
L. hamnosus GG—LGG
Daily adminis a ion
Dose: 5.2 ×107CFU/bw g/d.
Dilu ed in d inking wa e
T ea men leng h: 8 w.
↓Li e a accumula ion
↓Li e TG con en
↓Se um ALT le els
↓Li e in lamma ion
Down- egula ion o lipogenic
ma ke s in he li e :
↓Gene exp ession o Acc,Fas and Ch ebp.
Down- egula ion o p o-in lamma o y ma ke s and
media o s in he li e :
↓Gene exp ession o Il-1β,Il-8R,Tn αand Il-12.
Dec eased po al le els o LPS.
Up- egula ion o ma ke s o
in es inal mucosa in eg i y:
↑P o ein exp ession o
Occludin-1 and Claudin-1.
[16]Male C57BL/6 mice
4-week-old
HFD
(60% ene gy om a ).
L. hamnosus GG—LGG
O al daily adminis a ion
Dose: 1 ×108CFU/day
T ea men leng h: 13 w.
↓Li e weigh
↓Li e a accumula ion
↓Li e in lamma ion
Down- egula ion o lipogenic
ma ke s in he li e :
↓Gene exp ession o S ebp-1 and Ppa -γ.
Down- egula ion o genes ela ed o long-chain a y acid
up ake and lipop o ein syn hesis:
↓Gene exp ession o Cd36 and ApoB100.
Down- egula ion o p o-in lamma o y
ma ke s and media o s in he li e :
↓Gene exp ession o Il-6,Il-12,F4/80 and Cd11b.
Modula ion o gu mic obio a composi ion:
↑P opo ion o Bac e oide es.
[17]Male C57BL/6 mice
4-week-old HFD.
L. pa acasei N1115
O al daily adminis a ion Dose: 2.2 ×
109CFU/mL dilu ed in no mal saline
(0.5 mL/day).
T ea men leng h: 16 w.
↓Li e a accumula ion
↓Li e in lamma ion
↓Li e ib osis
Dec eased con en o hepa ic in lamma o y media o s
(Tn αand IL-1β).
Down- egula ion o p o-in lamma o y ma ke s and
media o s in he li e :
↓Gene exp ession o N -κB,Tl -4 and Lps.
Dec eased se um le els o li e
ib osis ma ke s (MAO).
Up- egula ion o ma ke s o
in es inal mucosa in eg i y:
↑P o ein exp ession o
Occludin-1 and Claudin-1.
[33]
Female
C57BL/6N mice
6–8-week-old
STD die wi h
30% uc ose in
d inking wa e .
L. hamnosus
O al daily ga age
Dose:1 ×109CFU/day.
T ea men leng h: 5 o 12 w.
↓Li e a accumula ion
↓Li e TG, TC and
VLDL con en
↓Li e in lamma ion
↓Li e apop o ic cells
Down- egula ion o li e inju y
p o ec ion ma ke s:
↑Gene exp ession o Fg 21.
Down- egula ion o p o-in lamma o y ma ke s and
media o s in he li e :
↓Gene exp ession o Tn αand Cxcl10.
Down- egula ion o lipogenic
ma ke s in he li e :
↓Gene exp ession o Fas,S ebp1c and Scd1.
↓P o ein exp ession o SREBP1c and ChREBP.
Up- egula ion o a y acid oxida ion
ma ke s in he li e :
↑Gene exp ession o CPT1 and PPARα.
Down- egula ion o ma ke s in ol ed
in hepa ic ce amide con en :
↓Me hyla ion o PP2AC.
In . J. Mol. Sci. 2022,23, 3167 5 o 25
Table 1. Con .
Re e ence Animal Model Expe imen al Condi ions P obio ic T ea men E ec s on Li e Mechanisms o Ac ion
[34]Male C57BL/6N mice
8-week-old
HFD/F (65% ene gy om
a and 30% uc ose solu ion).
L. plan a um NA136 g oup
O al daily adminis a ion
Dose: 1 ×109CFU/day.
T ea men leng h: 16 w.
↓Li e a accumula ion
↓Li e FFA con en
↓Li e in lamma ion
↓Li e ALT and AST le els
Down- egula ion o lipogenic ma ke s in he li e :
↓P o ein exp ession o FAS and SREBP-1.
↑Phospho yla ion o ACC.
Up- egula ion o ene gy yielding pa hways in he li e :
↑Phospho yla ion o AMPK.
Down- egula ion o oxida i e s ess in he li e :
↓Con en o MDA.
↑P o ein exp ession o HO-1 andN 2.
↑Con en o CAT.
↑Ac i i y o SOD.
[18]Male SPF C57BL/6J mice
6-week-old
No mal o Wes e n die
(42% ene gy om a ).
L. bulga icus
L. casei
L. hel e icus
P. pen osaceus KID7
Daily adminis a ion
Dose: 1 ×109CFU/g
suspended in dis illed wa e .
T ea men leng h: 8 w.
↓Li e s ea osis g ade
(all ea ed g oups)
↓Li e in lamma ion
(all ea ed g oups excep animals
ecei ing L. casei)
↓Li e /bw a io (g oups ea ed
wi h L. bulga icus,L. hel e icus
and P. pen osaceus)
↓Li e AST le els (g oups ea ed
wi h L. bulga icus and L. hel e icus)
↓Li e ALT le els (g oup ea ed
wi h L. bulga icus)
↓NAS (g oups ea ed wi h
L. bulga icus,L. hel e icus
and P. pen osaceus)
Down- egula ion o
mac ophage ma ke s in he li e :
↓Exp ession o Cd68 (g oups ea ed wi h
L. bulga icus,L. hel e icus,L. casei and P. pen osaceus).
Modula ion o gu mic obio a composi ion:
↓F/B a io (g oups ea ed wi h L. bulga icus,L. hel e icus,
P. pen osaceus and L. casei).
↑Con en o A. muciniphila (g oups ea ed wi h L. bulga icus,
L. hel e icus and L. casei).
Down- egula ion o p o-in lamma o y ma ke s and
media o s in he li e :
↓Gene exp ession o Tn αIl-6 and Il-1β
(in all he ea ed g oups).
[19]Male Swiss mice
4-week-old
HFD
(61% ene gy om a ).
B. longum
Daily o al ga age
Dose: 5 ×109CFU/kg bw/d
T ea men leng h: 4 w.
↓Li e lipid d ople size Up- egula ion o RAS ela ed genes in he li e :
↑Gene exp ession o Ace2 and Mas .
[20]Male C57BL/6N mice
6-week-old
HFD.
Animals also ecei ed
a 10% uc ose solu ion.
L. e men um—CQPC06
L. delb ueckii subsp.
Bulga icus—LDSB
Daily o al ga age
Dose o 1 ×109CFU/kg bw/d
(L. e men um—CQPC06) o
1×1010 CFU/kg bw/d
(L. e men um—CQPC06 and
L. delb ueckii subsp.
Bulga icus—LDSB) suspended
in s e ile saline.
T ea men leng h: 8 w.
↓Li e weigh and index
↓Li e TG
↓Se um and li e AST
and ALT le els
↓Se um and li e AKP le els
Dec eased ROS le els in he li e s o animals ecei ing
L. e men um—CQPC06 (a bo h doses) and L. delb ueckii
subsp. Bulga icus—LDSB.
Up- egula ion o a y acid oxida ion ma ke s in he li e
(L. e men um—CQPC06 (a bo h doses)):
↑Gene exp ession o Cp 1 and Ppa -α.
↑P o ein exp ession o CPT1 and PPAR-α.
Down- egula ion o lipogenic ma ke s in he li e
(L. e men um—CQPC06 (a bo h doses)):
↓Gene exp ession o C/ebp-αand Ppa -γ.
↓P o ein exp ession o C/EBP-αand PPAR-γ.
Up- egula ion o ma ke s o in es inal mucosa in eg i y
(L. e men um—CQPC06 (a bo h doses)):
↑P o ein exp ession o ZO-1, Occludin and Claudin-1.
Modula ion o gu mic obio a composi ion
(L. e men um—CQPC06 (a bo h doses) and L. delb ueckii
subsp. Bulga icus—LDSB):
↓F/B a io.
↑Con en o Akke mansia.
In . J. Mol. Sci. 2022,23, 3167 6 o 25
Table 1. Con .
Re e ence Animal Model Expe imen al Condi ions P obio ic T ea men E ec s on Li e Mechanisms o Ac ion
[21]Female
C57BL/6 mice
WSD
(40% ene gy om a ).
L. hamnosus GG ATCC 53103
L. lac is subsp. c emo is ATCC 19257
O al ga age
Th ice weekly
Dose: 1 ×109CFU
T ea men leng h: 16 w.
↓Li e weigh
(g oup ea ed wi h L. c emo is)
↓Li e PC, PS, TG and TG con en
(g oup ea ed wi h L. c emo is)
↓Li e lipid d ople a ea
(g oup ea ed wi h L. c emo is)
↓Li e in lamma ion
(g oup ea ed wi h L. c emo is)
Down- egula ion o hepa ic con en o lipids ela ed o
p o-in lamma o y esponse:
↓Le els o ARA con aining lipids
(g oup ea ed wi h L. c emo is).
Down- egula ion o in lamma ion associa ed
me aboli es in he li e :
↓Le els o Resol in E1, 9-HETE and 9HpODE
(g oup ea ed wi h L. c emo is).
[22]Male C57BL/6J mice
6-week-old
HFD
(45% ene gy om a ).
L. eu e i 6475
L. eu e i VPL3461
Daily o al ga age
(in a olume o 100 µL)
Dose o 1 ×109CFU/mL
T ea men leng h: 8 w.
↓Li e TG con en (all g oups) No speci ied.
[23]Male C57BL/6N mice
3–4-weeks-old
HFD
(42% ene gy om a ).
L. hamnosus GG
Daily adminis a ion
Dose: 1 ×108CFU/day mixed in he
expe imen al die .
T ea men leng h: 17 w.
↓Li e weigh
↓Li e TG con en
Modula ion o SCFA le els in he cecum:
↑Ace a e le els.
Modula ion o SCFA in he li e :
↑Ace a e le els.
Modula ion o an i-in lamma o y lipid media o le els:
↓´
ω6/ ´
ω3 PUFA a io.
[37]Male C57BL/6N mice
8-week-old
HFD/F
(65% ene gy om a and
30% die a y olume
p o ided as uc ose solu ion).
L. plan a um NA136
Daily o al
adminis a ion daily
Dose: 1 ×109CFU/day.
T ea men leng h: 16 w.
↓Li e lipid con en
Modula ion o gu mic obio a composi ion:
↑Bac e ial ichness and di e si y.
Up- egula ion o in es inal mucosa in eg i y ma ke s:
↑P o ein exp ession o igh -junc ion ma ke s
(ZO-1, Occludin, Claudin-1).
↓P o ein exp ession mucosal dys unc ion ma ke s (HIF-1α).
Dec eased se um le els o p o-in lamma o y cy okines
(TNF-α, IL-6, and IL-1β) and LPS.
Down- egula ion o p o-in lamma o y ma ke s and
media o s in he li e :
↓P o ein exp ession o NF-κB.
↓Phospho yla ion o p38.
[24]Male SPF C57BL/6J mice
6-week-old
WSD
(42% ene gy om a ).
L. acidophilus
L. e men um
L. pa acasei
L. plan a um
Daily adminis a ion
Dose: 1 ×109CFU
suspended in d inking wa e .
T ea men leng h: 8 w.
↓Li e s ea osis sco e (g oups ea ed
wi h L. pa acasei,L. plan a um and L.
acidophilus)
↓Li e TG con en (g oups ea ed
wi h L. acidophilus,L. e men um and
L. pa acasei)
Modula ion o mic obio a composi ion:
↑Bac e oide es con en (g oup ea ed wi h L. pa acasei).
↓Fi micu es con en (g oup ea ed wi h L. pa acasei).
[25]Male Sp ague-Dawley
a s HFD
L. acidophilus CGMCC 2106.
B. longum CGMCC 2107.
Daily adminis a ion
Dose: 1 ×1010 CFU/mL suspended
in d inking wa e .
T ea men leng h: 12 w.
↓Li e a accumula ion (g oup
ea ed wi h B. longum)
Modula ion o ecal mic obio a composi ion:
↑Bi idobac e ium con en (g oup ea ed wi h B. longum).
↑Lac obacillus con en (g oup ea ed wi h L. acidophilus).
In . J. Mol. Sci. 2022,23, 3167 7 o 25
Table 1. Con .
Re e ence Animal Model Expe imen al Condi ions P obio ic T ea men E ec s on Li e Mechanisms o Ac ion
[36] Male Fische 344 a s
CDAA die
(30% ene gy om a ).
Animals we e ed ad libi um and had
ee access o d inking wa e du ing
he whole expe imen .
C. bu y icum
Daily adminis a ion
Dose: 8.5 ×109CFU/g
mixed in he die .
T ea men leng h:
42 w.
↓Li e o al lipid and TG con en
↓Li e in lamma ion
↓NAFLD p og ession ( ib osis)
↓Se um ALT le els
↓Li e lipid pe oxida ion
↓Oxida i e s ess
Up- egula ion o ene gy yielding pa hways in he li e :
↑Phospho yla ion o AMPK.
Up- egula ion o a y acid oxida ion ma ke s in he li e :
↑P o ein exp ession o PPARα.
Down- egula ion o lipogenic ma ke s in he li e :
↓P o ein exp ession o SREBP-1c and PPAR-γ.
Down- egula ion o p o-in lamma o y ma ke s and
media o s in he li e :
↓P o ein exp ession o NF-kB and TNF-α.
Down- egula ion o
lipid pe oxida ion ma ke s in he li e :
↓Con en o 4-HNE and MDA.
Up- egula ion o an ioxidan ma ke s in he li e :
↑P o ein exp ession o N 2 and HO-1.
Up- egula ion o ma ke s o in es inal mucosa in eg i y:
↑P o ein exp ession o ZO1 and Ocln.
[26]Male Sp ague-Dawley
a s HFD.
L. plan a um NCU116-L
L. plan a um NCU116-H
Daily adminis a ion
Dose: 1 ×108CFU/mL
(L. plan a um NCU116-L) o
1×109CFU/mL
(L. plan a um NCU116-H) suspended
in a s e ile saline solu ion.
T ea men leng h: 5 w.
↓Li e AST le els
(g oup ea ed wi h L. plan a um
NCU116-H)
↓Li e oxida i e s ess
↓Li e TC and TG con en
↓Li e in lamma ion
Down- egula ion o oxida i e s ess ma ke s in li e :
↓MDA con en
(g oup ea ed wi h L. plan a um NCU116-H).
Up- egula ion o an ioxidan ma ke s in he li e :
↑Ac i i y o SOD and GPx (all g oups).
↑Ac i i y o CAT (g oup ea ed wi h
L. plan a um NCU116-H).
↑T-AOC (all g oups).
Down- egula ion o se um p o-in lamma o y cy okines:
↓Le els o LPS and IL-6 (all g oups).
↓Le els o TNFα(L. plan a um NCU116-H).
Up- egula ion o a y acid oxida ion and
lipolysis ma ke s in he li e :
↑Gene exp ession o Ppa α,Ppa γ,Ppa δ,Pgc1αand
Cp 1α(all g oups).
Down- egula ion o lipogenic ma ke s in he li e :
↓Gene exp ession o Fas,Acc and Scd1 (all g oups).
Modula ion o colonic mic obio a composi ion:
↓Gene exp ession o Bac e oides (all g oups).
↑Gene exp ession o Lac obacillus spp. and
Bi idobac e ium spp. (all g oups).
[24]Male Wis a
a s
HFD
(60% ene gy om a ).
L. pa acasei Jlus66
Daily o al adminis a ion
Doses: 1, 2 o 4 ×1010 CFU/d.
T ea men leng h: 20 w.
↓Li e weigh
↓Li e a accumula ion
↓Li e in lamma ion
↓Se um ALT le els (high dose)
No speci ied.
[35]
Male Sp ague-Dawley
Ra s
42-day-old
STD plus 20% uc ose
in d inking wa e .
L. acidophilus
B. coagulans
L. casei
L. eu e i
Daily adminis a ion
Dose: 1 ×109CFU/mL
suspended in d inking wa e .
T ea men leng h: 16 w.
↓Li e TG con en (g oups ea ed
wi h L. acidophilus and L. eu e i)
↓Se um ALT le els (all g oups)
↓Li e oxida i e s ess (all g oups)
Up- egula ion o an ioxidan esponse in he li e :
↑Con en o glu a hione (g oups ea ed wi h
L. acidophilus and L. casei).
↓Li e ROS o ma ion (g oups ea ed wi h L. acidophilus,
L. casei and B. coagulans).
↓Li e p o ein-ca bonyla ion (all g oups).
↓Li e lipid pe oxida ion (all g oups).
In . J. Mol. Sci. 2022,23, 3167 8 o 25
Table 1. Con .
Re e ence Animal Model Expe imen al Condi ions P obio ic T ea men E ec s on Li e Mechanisms o Ac ion
[28]Sp ague-Dawley a s
8-week-old
HFD
54% ene gy om a ).
Animals we e injec ed wi h 600
mg/kg/day o D-galac ose daily.
L. e men um DR9
L. plan a um DR7
L. eu e i 8513d
Daily adminis a ion
Dose: 1 ×1010 CFU/day dissol ed in
100 µL o saline and mixed in o 1 g o
expe imen al die .
T ea men leng h: 12 w.
↓Li e lipid con en (g oups ea ed
wi h L. e men um DR9, L. plan a um
DR7 and L. eu e i 8513d)
↓Li e in lamma ion (g oups ea ed
wi h L. e men um DR9, L. plan a um
DR7 and L. eu e i 8513d)
↓Li e ALP con en (g oups ea ed
wi h L. e men um DR9
and L. plan a um DR7)
Down- egula ion o lipogenic ma ke s in he li e :
↓Gene exp ession o Scd1 gene exp ession
(g oups ea ed wi h L. e men um DR9 and L. plan a um DR7).
Dec eased li e con en o p o-in lamma o y cy okines:
↓IL-4 le els (g oups ea ed wi h L. e men um DR9
and L. plan a um DR7).
Up- egula ion o ene gy yielding pa hways in he li e :
↑Gene exp ession o Ampkα1(g oups ea ed wi h
L. e men um DR9 and L. plan a um DR7) and Ampkα2gene
exp ession (g oup ea ed wi h L. plan a um DR7).
[16]
Male Wis a
Ra s
6-week-old
HFD
(45% o ene gy om a ).
Animals also ecei ed
10% uc ose in d inking wa e .
L. Plan a um s ain ATG-K2
L. Plan a um s ain ATG-K6
Daily o al ga age
Dose: 5 ×108CFU/d.
T ea men leng h: 8 w.
↓Li e TG and TC con en
↓Se um AST and ALT le els
(all g oups)
↓Se um ALP le els (all g oups)
↓Li e lipid pe oxida ion
Down- egula ion o lipogenic ma ke s in he li e :
↓Gene exp ession o S ebp-1c and Fas (all g oups).
↓P o ein exp ession o SREBP-1c
(g oup ea ed wi h L. Plan a um s ain ATG-K6).
↓P o ein exp ession o FAS (all g oups).
↓P o ein exp ession o C/EBP
(g oup ea ed wi h L. Plan a um s ain ATG-K2).
↑Phospho yla ion o ACC
(g oup ea ed wi h L. Plan a um s ain ATG-K2).
Up- egula ion o ene gy yielding pa hways in he li e :
↑Phospho yla ion o AMPK
(g oup ea ed wi h L. Plan a um s ain ATG-K2).
Up- egula ion o a y acid oxida ion ma ke s in he li e :
↑P o ein exp ession o CPT-1
(g oup ea ed wi h L. Plan a um s ain ATG-K2).
Dec eased li e MDA con en .
Modula ion o gu mic obio a composi ion:
↓Rela i e abundance o Fi micu es (all g oups).
↑Rela i e abundance o Bac e oide es (all g oups).
[30] Male Wis a a s HFD
(60% ene gy om a ).
B. animalis subsp. Lac is
V9
Daily o al ga age
Dose: 1 ×109CFU/mL.
T ea men leng h: 4 w.
↓Li e TG and FFA con en
↓Se um AST and ALT le els
↓Li e in lamma ion
↓P og ession o NASH
Down- egula ion o lipogenic ma ke s in he
li e :
↓Gene exp ession o S ebp-1c and Fas.
Up- egula ion o a y acid oxida ion ma ke s in he li e :
↑Gene exp ession o Ppa α.
Up- egula ion o ene gy yielding pa hways in he li e :
↑Phospho yla ion o AMPK.
Down- egula ion o NASH p og ession ma ke s in he li e :
↓Gene exp ession o Nl p3,Asc,Tl -4 and Tl -9.
Down- egula ion o p o-in lamma o y ma ke s and
media o s in he li e :
↓Gene exp ession o Tn α,IL-1βand IL-6.
↓Phospho yla ion o JNK, NF-kB, ERK and AKT.
In . J. Mol. Sci. 2022,23, 3167 9 o 25
Table 1. Con .
Re e ence Animal Model Expe imen al Condi ions P obio ic T ea men E ec s on Li e Mechanisms o Ac ion
[31]Male Sp ague-Dawley
a s
HFD
(45% ene gy om a ).
Eosinophil-Lac obacillus
Daily o al ga age
(312 mg/kg).
Dose: 1 ×107CFU/g.
T ea men leng h: 8 w.
↓Li e lipid con en
↓Li e in lamma ion
↓Se um and li e ALT
and AST le els
Modula ion o gu mic obio a composi ion:
↑Bac e ial di e si y.
↓Pa hogenic bac e ia con en .
Up- egula ion o li e lipogenesis inhibi o s:
↑P o ein exp ession o FGF15.
ACC: ace yl-CoA ca boxylase; ACE2: angio ensin-con e ing enzyme 2; Ak : p o ein kinase B; ALP: alkaline phospha ase; ALT: alanine ansaminase; AMPK: AMP-ac i a ed p o ein
kinase; ApoB100: apolipop o ein B100; ARA: a achidonic acid; ASC: Apop osis-associa ed speck-like p o ein con aining a caspase ec ui men domain; AST: aspa a e ansaminase;
bw: body weigh ; d: day; CAT: ca alase; CD36: clus e o di e en ia ion 36; CD11b: clus e o di e en ia ion molecule 11B; CD68: clus e o di e en ia ion 68; CDAA: choline-
de icien /L-amino acid-de ined; C/EBP-
α
: CCAAT/enhance binding p o ein
α
; CFU: colony- o ming uni ; ChREBP: ca bohyd a e- esponsi e elemen -binding p o ein; CPT1: ca ni ine
palmi oyl ans e ase 1; CXCL10: C-X-C Mo i Chemokine Ligand 10; ERK: ex acellula -signal- egula ed kinase; F4/80: EGF-like module-con aining mucin-like ho mone ecep o -like 1;
FAS: a y acid syn hase; F/B: Fi micu es/Bac e oide es; FGF15: ib oblas g ow h ac o -15; FGF21: ib oblas g ow h ac o -21; GPx: glu a hione pe oxidase; HFD: high- a die ; HFD/F:
high- a and uc ose die ; HIF-1
α
: hypoxia Inducible ac o 1 Subuni
α
; HO-1: heme oxygenase 1; IL-1
β
: in e leukin 1
β
; IL-4: in e leukin 4; IL-6: in e leukin 6; IL-8R: in e leukin 8
ecep o ; IL-12: in e leukin 12; JNK: janus kinase; LDL-c: LDL choles e ol; LPS: lipopolysaccha ide; MAO: monoamino oxidase; MASR: Mas ecep o ; MDA: malondialdehyde; NAS:
NAFLD ac i i y sco e; NASH: non-alcoholic s ea ohepa i is; NF-
κ
B: nuclea ac o kappa B; NLRP3: nod-like ecep o p o ein 3; N 2: nuclea ac o e y h oid 2– ela ed ac o 2; Ocln:
Occludin; p38: p38 MAP kinase; PBS: phospha e bu e ed saline; PGC1
α
: pe oxisome p oli e a o -ac i a ed ecep o gamma coac i a o 1-
α
; PP2AC: p o ein phospha ase 2 ca aly ic
subuni
α
; PPAR-
α
: pe oxisome p oli e a o -ac i a ed ecep o
α
; PPAR-
γ
: pe oxisome p oli e a o -ac i a ed ecep o
γ
; PPAR-
δ
: pe oxisome p oli e a o -ac i a ed ecep o
δ
; PUFA:
polyunsa u a ed a y acids; RAS: enin–angio ensin sys em; ROS: eac i e oxygen species; SCD1: s ea oyl-CoA desa u ase; SCFA: sho -chain a y acids; SOD: supe oxide dismu ase;
SPF: speci ic pa hogen- ee; SREBP-1: S e ol egula o y elemen -binding p o ein 1; STD: s anda d; T-AOC: o al an ioxidan capaci y; TC: o al choles e ol; TG: iglyce ides; TLR-4:
oll-like ecep o 4; TLR-9: oll-like ecep o 9; Tn
α
: umo nec osis ac o
α
; w: weeks; WSD: wes e n-s yle die ; ZO1: Zonula Occludens 1; 4-HNE: 4-hyd oxynonenal; 9-HETE:
9-hyd oxy-5Z,7E,11Z,14Z-eicosa e aenoic acid; 9HpODE: 9-hyd ope oxy-10E,12Z-oc adecadienoic acid; ↓: signi ican educ ion; ↑: signi ican inc ease.
In . J. Mol. Sci. 2022,23, 3167 16 o 25
much a en ion should be paid o he analysis and in e p e a ion o such da a, since
hese expe imen al condi ions a e no likely o be ep oduced in humans o ob ious
e hical easons.
3. E ec s o P obio ic Adminis a ion (Single S ain and Mix u es) in NAFLD
P e en ion: E idence om Clinical T ials
The po en ial o p obio ics o NAFLD managemen has also been in es iga ed in
humans. In his ega d, he a ailable s udies add essing he e ec s o p obio ics in NAFLD
a e mo e limi ed han hose ca ied ou in animals. Unlike p eclinical s udies, in which
he p e en i e e ec o p obio ics on s ea osis has been analyzed, s udies in humans ha e
add essed hei he apeu ic e ec s. In some s udies, p obio ics we e combined wi h o he
compounds/molecules, hus c ea ing a synbio ic, and consequen ly, he epo ed e ec s in
NAFLD ea men canno be a ibu ed solely o p obio ics. This ep esen ed a limi a ion
when selec ing sui able s udies o be included in his na a i e e iew a icle. Mo eo e ,
besides he p obio ic ea men , he pa icipan s also ecei ed some so o die a y ad ice
and/o we e encou aged o p ac ice physical ac i i y.
Acco ding o he majo i y o he clinical ials included in his e iew a icle, p obio ic
adminis a ion seems o be e ec i e in he ea men o NAFLD (Table 3). In e es ingly, and
con a y o ha obse ed in p eclinical s udies, mos o his esea ch has been conduc ed
using p obio ic s ain mix u es, ins ead o single-s ain p obio ics. Dec eased hepa ic lipid
con en , educed s ea osis g ade and lowe ed se um ansaminase le els ha e been epo ed
in s udies using di e en p obio ic s ain combina ions (including om 2 o 8 di e en
p obio ic bac e ia s ains), doses ( om 5
×
10
8
CFU/day o 22.5
×
10
10
CFU/day) and
adminis a ion pe iods ( om 8 weeks o 12 mon hs) in pa ien s ea u ing NAFLD [53–58].
I is wo h no ing ha hese e ec s we e desc ibed e en in s udies whe e all he pa icipan s
(including he con ol g oup) ecei ed die a y ad ice (aimed a inducing body weigh
educ ion), and also included physical ac i i y p og ams o pha maceu ical ea men o
u he heal h al e a ions (s a ins and ib a es) [
56
,
57
]. In addi ion, simila hepa op o ec i e
e ec s we e also desc ibed in a s udy in which p obio ics we e adminis e ed mixed in
a yogu , ins ead as a supplemen (capsule o sache ) [
59
]. Ne e heless, he e a e also
s udies in which he adminis a ion o p obio ics did no esul in he imp o emen o
ma ke s o li e inju y in pa ien s wi h NAFLD, despi e he ac ha he doses used and
he adminis a ion pe iods we e simila o hose s udies in which signi ican imp o emen s
we e epo ed (Table 3) [
60
,
61
]. In his ega d, acco ding o Mohamed No e al. [
60
],
he educed sample size, along wi h he appa en highe a iabili y o Malaysians’ gu
mic obio a composi ion (due o a mo e di e se die a y in ake), may ha e in luenced he
ob ained esul s. Mo eo e , he au ho s also poin ed o a change in die a y a obse ed
in he g oup ecei ing he p obio ic, which could ha e somehow blun ed he po en ial
bene icial e ec s o he p obio ic in e en ion [
60
]. As a as he s udy ca ied ou by
Chong e al. [
61
] is conce ned, di e ences in baseline cha ac e is ics be ween he p obio ic
and he placebo g oups, as well as he impossibili y o de e mine he pa icipan s’ NAFLD
se e i y, we e poin ed ou by he au ho s as po en ial ac o s in luencing he ou comes o
he s udy. Mo eo e , he pa icipan s in he s udies in which no p obio ic-adminis a ion-
de i ed bene i s in NAFLD we e epo ed we e olde han in he es o he s udies. Since
age- ela ed a ia ions in gu mic obio a composi ion ha e been iden i ied [
62
], i canno be
uled ou ha his a iable may ha e also in luenced he ou comes o hese s udies.
In . J. Mol. Sci. 2022,23, 3167 17 o 25
Table 3. S udies conduc ed in humans add essing he e ec s o di e en p obio ics (single s ain and mix u es) in NAFLD ea men (PICO o ma ).
Re e ence Popula ion In e en ion Compa ison Ou come
[53]
28 adul s wi h NAFLD
20 men
8 women.
Daily consump ion
P obio ic mix u e
(5 ×108CFU):
L. bulga icus
S. he mophilus
T ea men pe iod: 3 mon hs.
Placebo. ↓Se um ALT, AST and GGT le els.
[59]
72 obese adul s wi h NAFLD
33 men
39 women
Age: 23–63 yea s old.
BMI: 25–40 kg/m2
Daily consump ion
P obio ic-en iched
yogu (300 g/d):
L. bulga icus
S. he mophilus
L. acidophilus
(6.46 ×106CFU/g)
B. lac is Bb12 (4.97 ×106CFU/g)
T ea men pe iod: 8 weeks
Daily consump ion
Con en ional yogu
(300 g/d)
L. bulga icus
S. he mophilus
↓Se um ALT and AST le els.
[54]
64 obese adolescen s wi h NAFLD
Age: 10–18 yea s old
BMI >85 h pe cen ile (age and sex speci ic)
Daily consump ion
P obio ic mix u e
(1 capsule):
L. acidophilus ATCC B3208 (3 ×109CFU)
B. lac is DSMZ 32269 (6 ×109CFU)
B. bi idum ATCC SD6576 (2 ×109CFU)
L. hamnosus DSMZ 21690 (2 ×109CFU)
T ea men pe iod: 12 weeks
Placebo. ↓Se um ALT and AST le els.
↓Fa y li e g ade (sonog aphic g ading).
[55]
58 adul pa ien s wi h NAFLD and T2DM
Age: 18–65 yea s old
BMI > 25 kg/m2
Daily consump ion
P obio ic mix u e (1 sache o 10 g).
Lac obacillus + Lac ococcus (6 ×1010 CFU/g)
Bi idobac e ium (1 ×1010 CFU/g)
P opionibac e ium (3 ×1010 CFU/g)
Ace obac e (1 ×106CFU/g)
T ea men pe iod: 8 weeks
Placebo.
↓FLI and LS.
↓Se um AST and GGT le els.
Dec eased ci cula ing le els o
p oin lamma o y ma ke s:
↓Se um TNF-αand IL-6 le els.
[56]
65 obese adul s wi h NAFLD
33 men
32 women
Age: 19–75 yea s old
BMI >25 kg/m2
Mean hepa ic MRI-PDFF 16.2%
Daily consump ion
P obio ic mix u e (con aining 1 ×109CFU/1.4 g)
L. acidophilus CBT LA1
L. hamnosus CBT LR5 (human eces)
L. pa acasei CBT LPC5 (Ko ean e men ed ood—jeo gal)
P. pen osaceus CBT SL4 (Ko ean e men ed ege able
p oduc —kimchi)
B. lac is CBT BL3
B. b e e CBT BR3 (Ko ean in an eces)
T ea men pe iod: 12 weeks
Placebo.
↓IHF ac ion.
Modula ion o gu mic obio a composi ion:
↑Rela i e abundances o
L. acidophilus
L. hamnosus
P. pen osaceus
B. lac is
B. b e e
In . J. Mol. Sci. 2022,23, 3167 18 o 25
Table 3. Con .
Re e ence Popula ion In e en ion Compa ison Ou come
[57]30 adul s wi h NAFLD:
[ALT] and [AST] >1.5- old no mal le els
Daily consump ion
P obio ic mix u e
(2 capsules con aining 11.25 ×1010 CFU, each)
L. pa acasei DSM 24733,
L. plan a um DSM 24730
L. acidophilus
DSM 24735
L. delb ueckii subsp. Bulga icus DSM 24734
B. longum DSM 24736
B. in an is DSM 24737
B. b e e DSM 24732
S. he mophilus DSM 24731
T ea men pe iod: 12 mon hs
Placebo.
Imp o emen o li e his ology:
↓Hepa ocy e ballooning.
↓Lobula in lamma ion.
↓NAS sco e.
↓Se um ALT le els.
↓Se um ALP le els.
↓Se um p o-in lamma o y
cy okines: Il-1β, IL-6 and
TNF-α.
[58]
60 adul s wi h NAFLD 43 men
17 women
Age: 20–60 yea s old
BMI: 20–40 kg/m2
Daily consump ion
P obio ic mix u e (1 capsule con aining 5 ×109CFU)
L. casei
L. hamnosus
L. acidophilus
B. longum
B. b e e
T ea men pe iod: 12 weeks
Placebo. ↓Se um ALT, AST and GGT le els
↓Se um ALP le els.
[60]
35 adul s wi h NAFLD 28 men
7 women
Age: 25–70 yea s old
Mean BMI 32.6 ±5.0 kg/m2
Daily consump ion
P obio ic mix u e (2 sache s o VSL#3 wice daily)
S. he mophilus
B. b e e
B. in an is
B. longum
L. acidophilus
L. plan a um
L. pa acasei
L delb ueckii subsp. bulga icus
T ea men pe iod: 10 weeks
Placebo. No signi ican imp o emen s in
ma ke s o li e inju y.
[61]
39 obese adul s
28 men
11 women wi h NAFLD:
Fa y li e sco e >263 dB/m
Daily consump ion
P obio ic mix u e (1 sache con aining 3 ×1010 CFU
wice daily)
L. acidophilus BCMC 12,130 (107 mg)
L. lac is MCMC 12,451 (107 mg)
B. bi idum BCMC 02290 (107 mg)
B. in an is BCMC 02129 (107 mg)
B. longum BCMC 02120 (107 mg)
T ea men pe iod: 6 mon hs
Placebo.
No signi ican imp o emen s
in li e ib osis pa ame e s o se um ma ke s o
in lamma ion.
ALP: alkaline phospha ase; ALT: alanine ansaminase; AST: aspa a e amino ans e ase; BMI: body mass index; CFU: colony- o ming uni s; FLI: a y li e index; GGT:
γ
glu amyl
ans e ase; HbA1c: glyca ed hemoglobin; IHF: in ahepa ic a ; IL-1
β
: in e leukin 1
β
; IL-6: in e leukin 6; m: men; LS: li e s i ness; MRI-PDFF: magne ic- esonance-imaging-
de i ed p o on densi y a ac ion; NAFLD: non-alcoholic a y li e disease; NAS: NAFLD ac i i y sco e; TNF-
α
: umo nec osis ac o
α
; w: women;
↓
: signi ican educ ion;
↑: signi ican inc ease.
In . J. Mol. Sci. 2022,23, 3167 19 o 25
Besides he a o emen ioned e ec s on li e TG accumula ion, an i-in lamma o y p op-
e ies ha e also been desc ibed in s udies add essing he e ec s o p obio ic adminis a ion
in NAFLD pa ien s. In his ega d, dec eased ci cula ing le els o p o-in lamma o y me-
dia o s such as TNF-
α
, IL-1
β
o IL-6, as well as lowe ed hepa ocy e ballooning and li e
lobula ib osis (assessed by his ological analysis) ha e been obse ed [
55
,
57
]. Simila ly,
and in line wi h he ou comes ound in p eclinical s udies, p obio ic adminis a ion also e-
sul ed in gu mic obio a modula ion in pa ien s wi h NAFLD. In his case, inc eased ela i e
abundances o L. acidophilus,L. hamnosus,P. pen osaceus,B. lac is and B. b e e we e ound
in obese NAFLD pa ien s ecei ing a p obio ic mix u e (con aining six di e en s ains)
o 12 weeks [
56
]. By con as , none o he s udies included in his e iew sec ion ha e
add essed he e ec s o p obio ic adminis a ion on SCFA le els and/o in es inal in eg i y.
Based on he esul s epo ed in clinical ials, i could be concluded ha in gene al
e ms, p obio ic adminis a ion e ec i ely imp o es ma ke s o li e inju y in pa ien s
wi h NAFLD. Indeed, he majo e ec s ha ha e been desc ibed o da e, such as lowe
in ahepa ic lipid con en , dec eased li e inju y, as well as dec eased ci cula ing ansami-
nase and p o-in lamma o y cy okine le els, a e compa ible o hose epo ed in p eclinical
s udies. Howe e , one o he main limi a ions o s udies conduc ed in humans elies on
he di icul y o ob ain samples ha may make i possible o in es iga e he mechanisms
o ac ion in ol ed in hese hepa op o ec i e e ec s. No wi hs anding ha di e en non-
in asi e imaging echniques including ul asound, compu e omog aphy o magne ic
esonance imaging ha e been demons a ed o be e ec i e de ec ing li e a in il a ion,
hese a e no app op ia e o assess li e in lamma ion o ib osis [
63
]. Fu he mo e, hese
echniques a e no sui able o explo e he pa hways and mechanisms o ac ion modula ed
by p obio ic adminis a ion. In his ega d, li e biopsies ep esen he gold s anda d o
s udy hepa ic in lamma ion and ib osis, as well as he mechanisms o ac ion in ol ed in
he e ec s media ed by p obio ics. Un o una ely, since he p ocedu es needed o ob ain
such samples happen o be e y in asi e, ma ke s ha can be mo e easily s udied (such as
se um ansaminase o cy okine le els) a e usually selec ed o elucida e he e ec i eness o
hese app oaches in NAFLD.
4. Limi a ions o P obio ic Adminis a ion and Po en ial Al e na i es
Acco ding o he s udies included in his e iew, as well he ones ound in he li e a u e
ha add ess he e ec s o p obio ics on diseases o he han NAFLD, p obio ic adminis a-
ion ep esen s an e ec i e he apeu ic ool o he managemen o an a ay o me abolic
al e a ions including obesi y, diabe es o dyslipidemia [
64
–
68
]. In his ega d, besides he
mo e “con en ional” p obio ics such as Bi idobac e ium and Lac obacillus, much a en ion
has also been paid o o he mic oo ganisms e e ed o as nex -gene a ion p obio ics (NGP)
as po en ial he apeu ic app oaches o NAFLD managemen . These “new” p obio ics,
esul ing om imp o ed cul u e me hods, bioin o ma ics and nex -gene a ion sequencing,
include such species as Akke mansia muciniphila,Faecalibac e ium p ausni zii,Eubac e ium
hallii,P opionibac e ium,Bac e oides agilis and genus belonging o he Clos idia clus e s
IV, XIVa and XVIII [
69
]. Fo ins ance, lowe Akke mansia muciniphila abundances ha e
been ela ed o me abolic diso de s such as obesi y and NAFLD [
70
,
71
]. In e es ingly, he
adminis a ion o his bac e ia was ound o be e ec i e in amelio a ing obesi y and ela ed
me abolic diso de s, bu wi hou a ec ing gu mic obio a composi ion [
65
]. Mo eo e , he
adminis a ion o hea - ea ed Akke mansia muciniphila was also shown o exe me abolic
bene i s, simila o hose p oduced by he adminis a ion o iable bac e ia [
72
]. In addi ion,
i was epo ed ha in NAFLD pa ien s, he abundance o Faecalibac e ium p ausni zii ends
o be low. Since his bac e ia is known o p oduce bu y a e, i s use ulness o NAFLD
p e en ion was p oposed [
69
]. Simila ly, Rosebu ia spp. a e bu y a e-p oducing bac e ia,
and as in he case o Faecalibac e ium p ausni zii, may be e ec i e o NAFLD manage-
men . Indeed, i was epo ed ha Rosebu ia spp. adminis a ion educes hepa ic s ea osis
and in lamma ion, mainly by es o ing he gu mic obio a en i onmen and in es inal
In . J. Mol. Sci. 2022,23, 3167 20 o 25
in eg i y [
73
]. The e o e, cu en a ailable da a sugges ha he NGP may ep esen an
addi ional he apeu ic ool o NAFLD p e en ion and ea men .
Despi e he a o emen ioned p obio ic heal h bene i s, hei adminis a ion also in-
ol es some haza ds, since his he apeu ic app oach is based on he adminis a ion o
li e/ iable mic oo ganisms o ulne able subjec s, which in u n esul s in an inc eased
isk o sys emic in ec ion and/o immune sys em o e s imula ion [
74
]. In his line, much
a en ion has been paid o he usage o pa abio ics and pos bio ics as al e na i e app oaches
o p obio ics [75].
In he case o pa abio ics, also e e ed o as pa ap obio ics o ghos p obio ics, hese
a e usually ob ained by inac i a ion o p obio ic bac e ia, mainly by means o he mic
ea men [
76
]. In his ega d, he e icacy o pa abio ics elies on he molecules and
compounds con ained in inac i a ed bac e ial cells, and no in hei iabili y [
77
]. In
compa ison o p obio ics (li e bac e ia), pa abio ics ep esen se e al po en ial ad an ages,
which include a lowe isk o in ec ion and an ibio ic esis ance acquisi ion/ ans e , as well
as an easie s o age and handling [
78
]. E en hough he a ailable da a ega ding he usage
o pa abio ics in he managemen o di e en diseases a e s ill sca ce, i was desc ibed
ha adminis a ion o hea -inac i a ed p obio ic bac e ia (S ep ococcus he mophilus MN-
ZLW-002) is e ec i e in p e en ing high- a die eeding induced body weigh gain, insulin
esis ance and dyslipidemia in mice [
79
]. In addi ion, acco ding o da a epo ed in clinical
ials, he con inuous adminis a ion o agmen ed Lac obacillus amylo o us CP1563 (hea
inac i a ed, lyophilized and hen milled) o 12 weeks signi ican ly educes whole body
and isce al a , amelio a es ma ke s ela ed o glycaemic con ol ( educed as ing blood
glucose and insulin le els) and imp o es dyslipidemia ( educing blood TG, and o al
and LDL choles e ol le els) in subjec s ea u ing class I obesi y [
80
]. Indeed, in a ecen
sys ema ic e iew add essing he e icacy o pa abio ics in he p e en ion and ea men
o di e en diseases, compa ed o p obio ics, no signi ican di e ences we e epo ed
ega ding he e ec i eness o pa abio ics in he majo i y o he p e en i e and ea men
ials analyzed (86% and 69%, espec i ely) [
81
]. Thus, al hough he a ailable e idence
conce ning pa abio ic use as he apeu ic app oach is limi ed, he esul s epo ed so a
sugges s ha o e all, hei e icacy is simila o ha a ibu ed o p obio ics.
In espec o pos bio ics, hese encompasses a wide spec um o non- iable bac e ial
p oduc s and cell componen s wi h po en ial bioac i e ac i i y in he hos , including ce ain
i amins (A and K, o ins ance), bile acids, SCFAs, polyamines, b anched-chain amino
acids o componen s o bac e ial cell wall, such as eichoic acids [
77
,
82
]. As occu s o
pa abio ics, da a ega ding he e icacy o pos bio ics is s ill scan and mainly limi ed o
p eclinical s udies. Acco ding o he s udies ha ha e been published so a , in olde
mice, he adminis a ion o lipo eichoic acid om hea -inac i a ed
Lac obacillus pa acasei
D3-5 p e en ed high- a die eeding-induced me abolic dys unc ion [
83
]. Simila ly, he
adminis a ion o he polyamine spe midine has been epo ed o e ec i ely p e en
high- a die eeding-induced body weigh gain, li e lipid accumula ion o insulin esis-
ance in mice [
84
,
85
]. In hese cases, he adminis a ion o he pos bio ics esul ed in he
amelio a ion o gu mic obio a dysbiosis and in lamma ion, as well as in he eco e y o
in es inal in eg i y [83–85].
Al oge he , and despi e he ac ha u he esea ch is wa an ed, hese da a sugges
ha he adminis a ion o pa abio ics and pos bio ics may also p o e e ec i e in he
managemen o ce ain diseases, as well as highligh ing ha he unc ionali y o such
compounds is beyond mic obial iabili y.
5. Conclusions
The aim o he p esen na a i e e iew a icle was o summa ize he e idence a ailable
ega ding he e ec i eness o p obio ic adminis a ion in NFALD managemen . In his
con ex , s udies conduc ed in oden models ha e e ealed ha bo h he adminis a ion o
single-s ain p obio ics, as well as he adminis a ion o p obio ic mix u es, ep esen an
e ec i e app oach in he p e en ion o his li e condi ion. Based on he magni ude o he
In . J. Mol. Sci. 2022,23, 3167 21 o 25
obse ed e ec s, along wi h he desc ibed mechanisms o ac ion, i could be sugges ed ha
compa ed o he usage o single-s ain p obio ics, he combina ion o di e en p obio ic
s ains does no ep esen an ad an age. In he case o s udies conduc ed in humans, in he
majo i y o hem, p obio ic adminis a ion also esul ed in he amelio a ion o ma ke s o
li e inju y in NAFLD pa ien s.
Wi h ega d o he mechanisms o ac ion unde lying he e ec s ha ha e been de-
sc ibed so a , p eclinical s udies ha e demons a ed ha p obio ics (single o mixed
s ains) ac in he li e , down- egula ing lipid syn hesis, ac i a ing lipid oxida ion and
down- egula ing p o-in lamma o y pa hways, as well as in he gu , modula ing mic obio a
composi ion, in es inal in eg i y and he p oduc ion o mic obial me aboli es. As o clinical
s udies, da a desc ibing such mechanisms a e sca ce, mainly due o limi a ions in e ms
o ob aining samples. In his ega d, me agenomics and me abolomics may ep esen a
use ul ool o be e assess he e ec s o p obio ic adminis a ion using samples such as
blood, u ine o eces. Simila ly, u he esea ch is wa an ed in o de o elucida e whe he
he adminis a ion o pa abio ics o pos bio ics cons i u es a eal al e na i e o he usage o
p obio ics o NAFLD managemen , and hus, o o e come he limi a ion ha ep esen s
he adminis a ion o iable mic oo ganisms o ulne able subjec s.
Au ho Con ibu ions:
Concep ualiza ion, I.M.-L., L.A.-G., J.A.M. and M.P.P.; w i ing—o iginal d a
p epa a ion, I.M.-L., L.A.-G. and M.P.P.; w i ing— e iewing and edi ing, I.M.-L., L.A.-G., J.A.M. and
M.P.P.; supe ision, J.A.M. and M.P.P.; unding acquisi ion, M.P.P. All au ho s ha e ead and ag eed
o he published e sion o he manusc ip .
Funding:
This esea ch was unded by CIBEROBN unde G an CB12/03/30007 and he Communi y
o Mad id unde G an Y2020/BIO-6600.
Ins i u ional Re iew Boa d S a emen : No applicable.
In o med Consen S a emen : No applicable.
Da a A ailabili y S a emen : No applicable.
Acknowledgmen s:
Iñaki Mil on-Laskiba acknowledges inancial suppo om he Juan de la
Cie a P og amme-T aining G an s o he Spanish S a e Resea ch Agency o he Spanish Minis e io
de Ciencia e Inno ación y Minis e io de Uni e sidades (FJC2019-038925-I).
Con lic s o In e es : The au ho s decla e no con lic o in e es .
Re e ences
1. Byass, P. The global bu den o li e disease: A challenge o me hods and o public heal h. BMC Med. 2014,12, 159. [C ossRe ]
2.
Engin, A. Non-Alcoholic Fa y Li e Disease. In Obesi y and Lipo oxici y; Engin, A.B., Engin, A., Eds.; Sp inge In e na ional
Publishing: Cham, Swi ze land, 2017; pp. 443–467.
3. Younossi, Z.; Ans ee, Q.M.; Ma ie i, M.; Ha dy, T.; Hen y, L.; Eslam, M.; Geo ge, J.; Bugianesi, E. Global bu den o NAFLD and
NASH: T ends, p edic ions, isk ac o s and p e en ion. Na . Re . Gas oen e ol. Hepa ol. 2018,15, 11–20. [C ossRe ] [PubMed]
4. Angulo, P. Nonalcoholic a y li e disease. N. Engl. J. Med. 2002,346, 1221–1231. [C ossRe ] [PubMed]
5.
B un , E.M.; Wong, V.W.S.; Nobili, V.; Day, C.P.; Sookoian, S.; Mahe , J.J.; Bugianesi, E.; Si lin, C.B.; Neuschwande -Te i, B.A.;
Rinella, M.E. Nonalcoholic a y li e disease. Na . Re . Dis. P ime s 2015,1, 15080. [C ossRe ] [PubMed]
6.
Buzze i, E.; Pinzani, M.; Tsocha zis, E.A. The mul iple-hi pa hogenesis o non-alcoholic a y li e disease (NAFLD). Me abolism
2016,65, 1038–1048. [C ossRe ] [PubMed]
7.
Jiang, X.; Zheng, J.; Zhang, S.; Wang, B.; Wu, C.; Guo, X. Ad ances in he In ol emen o Gu Mic obio a in Pa hophysiology o
NAFLD. F on . Med. 2020,7, 361. [C ossRe ] [PubMed]
8.
Gómez-Zo i a, S.; Agui e, L.; Mil on-Laskiba , I.; Fe nández-Quin ela, A.; T epiana, J.; Kaja abille, N.; Mosqueda-Solís, A.;
González, M.; Po illo, M.P. Rela ionship be ween changes in mic obio a and li e s ea osis induced by high- a eeding—A
e iew o oden models. Nu ien s 2019,11, 2156. [C ossRe ]
9.
So ic, S.; Cohen, D.E.; Kahn, C.R. Role o Die a y F uc ose and Hepa ic De No o Lipogenesis in Fa y Li e Disease. Dig. Dis. Sci.
2016,61, 1282–1293. [C ossRe ] [PubMed]
10.
Lian, C.; Zhai, Z.; Li, Z.; Wang, L. High a die - igge ed non-alcoholic a y li e disease: A e iew o p oposed mechanisms.
Chem.-Biol. In e ac . 2020,330, 109199. [C ossRe ]
In . J. Mol. Sci. 2022,23, 3167 22 o 25
11.
La son-Meye , D.E.; Newcome , B.R.; Heilb onn, L.K.; Volau o a, J.; Smi h, S.R.; Al onso, A.J.; Le e e, M.; Rood, J.C.;
Williamson, D.A.
; Ra ussin, E.; e al. E ec o 6-mon h calo ie es ic ion and exe cise on se um and li e lipids and ma ke s o
li e unc ion. Obesi y 2008,16, 1355–1362. [C ossRe ]
12.
Elias, M.C.; Pa ise, E.R.; de Ca alho, L.; Szejn eld, D.; Ne o, J.P. E ec o 6-mon h nu i ional in e en ion on non-alcoholic a y
li e disease. Nu i ion 2010,26, 1094–1099. [C ossRe ] [PubMed]
13.
Di ella, R.; Mazzocca, A.; Daniele, A.; Sabbà, C.; Pa adiso, A. Obesi y, Nonalcoholic Fa y Li e Disease and Adipocy o-kines
Ne wo k in P omo ion o Cance . In . J. Biol. Sci. 2019,15, 610–616. [C ossRe ] [PubMed]
14. Vandenplas, Y.; Huys, G.; Daube, G. P obio ics: An upda e. J. Pedia . 2015,91, 6–21. [C ossRe ] [PubMed]
15.
Pe umpail, B.J.; Li, A.A.; John, N.; Sallam, S.; Shah, N.D.; Kwong, W.; Cholanke il, G.; Kim, D.; Ahmed, A. The The apeu ic
Implica ions o he Gu Mic obiome and P obio ics in Pa ien s wi h NAFLD. Diseases 2019,7, 27. [C ossRe ]
16.
Kim, B.; Pa k, K.; Ji, Y.; Pa k, S.; Holzap el, W.; Hyun, C. P o ec i e e ec s o Lac obacillus hamnosus GG agains dyslipidemia in
high- a die -induced obese mice. Biochem. Biophys. Res. Commun. 2016,473, 530–536. [C ossRe ]
17.
Yao, F.; Jia, R.; Huang, H.; Yu, Y.; Mei, L.; Bai, L.; Ding, Y.; Zheng, P. E ec o Lac obacillus pa acasei N1115 and uc ooligosaccha ides
in nonalcoholic a y li e disease. A ch. Med. Sci. 2019,15, 1336–1344. [C ossRe ]
18.
Lee, N.Y.; Yoon, S.J.; Han, D.H.; Gup a, H.; Youn, G.S.; Shin, M.J.; Ham, Y.L.; Kwak, M.J.; Kim, B.Y.; Yu, J.S.; e al. Lac obacillus and
Pediococcus amelio a e p og ession o non-alcoholic a y li e disease h ough modula ion o he gu mic obiome. Gu Mic obes
2020,11, 882–899. [C ossRe ]
19.
Machado, A.S.; Oli ei a, J.R.; Lelis, D.D.F.; de Paula, A.M.B.; Guima ães, A.L.S.; And ade, J.M.O.; B andi, I.V.; San os, S.H.S.
O al P obio ic Bi idobac e ium Longum Supplemen a ion Imp o es Me abolic Pa ame e s and Al e s he Exp ession o he
Renin-Angio ensin Sys em in Obese Mice Li e . Biol. Res. Nu s. 2021,23, 100–108. [C ossRe ]
20.
Mu, J.; Tan, F.; Zhou, X.; Zhao, X. Lac obacillus e men um CQPC06 in na u ally e men ed pickles p e en s non-alcoholic a y
li e disease by s abilizing he gu -li e axis in mice. Food Func . 2020,11, 8707–8723. [C ossRe ]
21.
Naudin, C.R.; Mane -Smi h, K.; Owens, J.A.; Wynn, G.M.; Robinson, B.S.; Ma hews, J.D.; Reedy, A.R.; Luo, L.; Wol a h, A.A.;
Da by, T.M.; e al. Lac ococcus lac is Subspecies c emo is Elici s P o ec ion Agains Me abolic Changes Induced by a Wes e n-S yle
Die . Gas oen e ology 2020,159, 639–651.e5. [C ossRe ]
22.
Oh, J.; Schuele , K.L.; S aple on, D.S.; Alexande , L.M.; Yen, C.E.; Kelle , M.P.; A ie, A.D.; an Pijke en, J.P. Sec e ion o
Recombinan In e leukin-22 by Enginee ed Lac obacillus eu e i Reduces Fa y Li e Disease in a Mouse Model o Die -Induced
Obesi y. mSphe e 2020,5, e00183-20. [C ossRe ]
23.
Yau, Y.F.; El-nezami, H.; Galano, J.; Kundi, Z.M.; Du and, T.; Lee, J.C. Lac obacillus hamnosus GG and Oa Be a-Glucan Regula ed
Fa y Acid P o iles along he Gu -Li e -B ain Axis o Mice Fed wi h High Fa Die and Demons a ed An ioxidan and An i-
In lamma o y Po en ials. Mol. Nu . Food Res. 2020,64, 2000566. [C ossRe ]
24.
Lee, N.Y.; Shin, M.J.; Youn, G.S.; Yoon, S.J.; Choi, Y.R.; Kim, H.S.; Gup a, H.; Han, S.H.; Kim, B.K.; Lee, D.Y.; e al. Lac obacillus
a enua es p og ession o nonalcoholic a y li e disease by lowe ing choles e ol and s ea osis. Clin. Mol. Hepa ol.
2021
,
27, 110–124. [C ossRe ]
25.
Xu, R.; Wan, Y.; Fang, Q.; Lu, W.; Cai, W. Supplemen a ion wi h p obio ics modi ies gu lo a and a enua es li e a accumula ion
in a nonalcoholic a y li e disease model. J. Clin. Biochem. Nu . 2012,50, 72–77. [C ossRe ]
26.
Li, C.; Nie, S.; Zhu, K.; Ding, Q.; Li, C.; Xiong, T.; Xie, M. Lac obacillus plan a um NCU116 imp o es li e unc ion, oxida i e
s ess and lipid me abolism in a s wi h high a die induced non-alcoholic a y li e disease. Food Func .
2014
,5, 3216–3223.
[C ossRe ] [PubMed]
27.
Ye, H.; Li, Q.; Zhang, Z.; Sun, M.; Zhao, C.; Zhang, T. E ec o a no el po en ial p obio ic Lac obacillus pa acasei Jlus66 isola ed
om e men ed milk on nonalcoholic a y li e in a s. Food Func . 2017,8, 4539–4546. [C ossRe ] [PubMed]
28.
Lew, L.-C.; Ho , Y.-Y.; Jaa a , M.-H.; Lau, A.-S.-Y.; Lee, B.-K.; Chuah, L.-O.; Yap, K.-P.; Azlan, A.; Azzam, G.; Choi, S.-B.; e al.
Lac obacillus S ains Alle ia ed Hype lipidemia and Li e S ea osis in Aging Ra s ia Ac i a ion o AMPK. In . J. Mol. Sci.
2020
,
21, 5872. [C ossRe ] [PubMed]
29.
Pa k, E.-J.; Lee, Y.-S.; Kim, S.M.; Pa k, G.-S.; Lee, Y.H.; Jeong, D.Y.; Kang, J.; Lee, H.-J. Bene icial E ec s o Lac obacillus plan a um
S ains on Non-Alcoholic Fa y Li e Disease in High Fa /High F uc ose Die -Fed Ra s. Nu ien s 2020,12, 542. [C ossRe ]
30.
Yan, Y.; Liu, C.; Zhao, S.; Wang, X.; Wang, J.; Zhang, H.; Wang, Y.; Zhao, G. P obio ic Bi idobac e ium lac is V9 a enua es hepa ic
s ea osis and in lamma ion in a s wi h non-alcoholic a y li e disease. AMB Exp ess 2020,10, 101. [C ossRe ]
31.
Luo, M.; Yan, J.; Wu, L.; Wu, J.; Chen, Z.; Jiang, J.; Chen, Z.; He, B. P obio ics Alle ia ed Nonalcoholic Fa y Li e Disease in
High-Fa Die -Fed Ra s ia Gu Mic obio a/FXR/FGF15 Signaling Pa hway. J. Immunol. Res. 2021,2021, 2264737. [C ossRe ]
32.
Ri ze, Y.; Bá dos, G.; Claus, A.; Eh mann, V.; Be gheim, I.; Schwie z, A.; Bischo , S.C. Lac obacillus hamnosus GG P o ec s agains
Non-Alcoholic Fa y Li e Disease in Mice. PLoS ONE 2014,9, e80169. [C ossRe ]
33.
Zhao, C.; Liu, L.; Liu, Q.; Li, F.; Zhang, L.; Zhu, F.; Shao, T.; Ba e, S.; Chen, Y.; Li, X.; e al. Fib oblas g ow h ac o 21 is equi ed
o he he apeu ic e ec s o Lac obacillus hamnosus GG agains uc ose-induced a y li e in mice. Mol. Me ab.
2019
,29, 145–157.
[C ossRe ]
34.
Zhao, Z.; Wang, C.; Zhang, L.; Zhao, Y.; Duan, C.; Zhang, X.; Gao, L.; Li, S. Lac obacillus plan a um NA136 imp o es he
non-alcoholic a y li e disease by modula ing he AMPK/N 2 pa hway. Appl. Mic obiol. Bio echnol.
2019
,103, 5843–5850.
[C ossRe ]
In . J. Mol. Sci. 2022,23, 3167 23 o 25
35.
Aza ang, A.; Fa shad, O.; Omma i, M.M.; Jamshidzadeh, A.; Heida i, R.; Aboo alebi, S.N.; Gholami, A. P o ec i e Role o
P obio ic Supplemen s in Hepa ic S ea osis: A Ra Model S udy. Biomed. Res. In . 2020,2020, 5487659. [C ossRe ]
36.
Endo, H.; Niioka, M.; Kobayashi, N.; Tanaka, M.; Wa anabe, T. Bu y a e-P oducing P obio ics Reduce Nonalcoholic Fa y Li e
Disease P og ession in Ra s: New Insigh in o he P obio ics o he Gu -Li e Axis. PLoS ONE 2013,8, e63388. [C ossRe ]
37.
Zhao, Z.; Chen, L.; Zhao, Y.; Wang, C.; Duan, C.; Yang, G.; Niu, C.; Li, S. Lac obacillus plan a um NA136 amelio a es nonalco-
holic a y li e disease by modula ing gu mic obio a, imp o ing in es inal ba ie in eg i y, and a enua ing in lamma ion.
Appl. Mic obiol. Bio echnol. 2020,104, 5273–5282. [C ossRe ] [PubMed]
38.
Rakhshandeh oo, M.; Knoch, B.; Mülle , M.; Ke s en, S. Pe oxisome P oli e a o -Ac i a ed Recep o Alpha Ta ge Genes. PPAR Res.
2010,2010, 612089. [C ossRe ]
39.
Can ó, C.; Auwe x, J. PGC-1alpha, SIRT1 and AMPK, an ene gy sensing ne wo k ha con ols ene gy expendi u e. Cu . Opin.
Lipidol. 2009,20, 98–105. [C ossRe ]
40.
A ulsel an, P.; Fa d, M.T.; Tan, W.S.; Go hai, S.; Faku azi, S.; No haizan, M.E.; Kuma , S.S. Role o An ioxidan s and Na u al
P oduc s in In lamma ion. Oxid. Med. Cell. Longe . 2016,2016, 5276130. [C ossRe ] [PubMed]
41.
Jin, C.J.; Sellmann, C.; Engs le , A.J.; Ziegenha d , D.; Be gheim, I. Supplemen a ion o sodium bu y a e p o ec s mice om he
de elopmen o non-alcoholic s ea ohepa i is (NASH). B . J. Nu . 2015,114, 1745–1755. [C ossRe ] [PubMed]
42.
Chambe s, E.S.; P es on, T.; F os , G.; Mo ison, D.J. Role o Gu Mic obio a-Gene a ed Sho -Chain Fa y Acids in Me abolic and
Ca dio ascula Heal h. Cu . Nu . Rep. 2018,7, 198–206. [C ossRe ] [PubMed]
43.
Dangana, E.O.; Omolekulo, T.E.; A eola, E.D.; Olaniyi, K.S.; Soladoye, A.O.; Ola unji, L.A. Sodium ace a e p o ec s agains
nico ine-induced excess hepa ic lipid in male a s by supp essing xan hine oxidase ac i i y. Chem.-Biol. In e ac .
2020
,316, 108929.
[C ossRe ]
44.
Kim, B.; Kwon, J.; Kim, M.; Pa k, H.; Ji, Y.; Holzap el, W.; Hyun, C. P o ec i e e ec s o Bacillus p obio ics agains high- a
die -induced me abolic diso de s in mice. PLoS ONE 2018,13, e0210120. [C ossRe ] [PubMed]
45.
Kim, H.; Kim, J.; Kim, J.; Jang, S.; Han, M.J.; Kim, D.H. Lac obacillus plan a um LC27 and Bi idobac e ium longum LC67 simul-
aneously alle ia e high- a die -induced coli is, endo oxemia, li e s ea osis, and obesi y in mice. Nu . Res.
2019
,67, 78–89.
[C ossRe ]
46.
Ahmadi, S.; Wang, S.; Nagpal, R.; Wang, B.; Jain, S.; Razazan, A.; Mish a, S.P.; Zhu, X.; Wang, Z.; Ka anagh, K.; e al. A human-
o igin p obio ic cock ail amelio a es aging- ela ed leaky gu and in lamma ion ia modula ing he mic obio a/ au ine/ igh
junc ion axis. JCI Insigh 2020,5, e132055. [C ossRe ] [PubMed]
47.
Li, H.; Liu, F.; Lu, J.; Shi, J.; Guan, J.; Yan, F.; Li, B.; Huo, G. P obio ic Mix u e o Lac obacillus plan a um S ains Imp o es Lipid
Me abolism and Gu Mic obio a S uc u e in High Fa Die -Fed Mice. F on . Mic obiol. 2020,11, 512. [C ossRe ] [PubMed]
48.
Jiang, J.; Xiong, J.; Ni, J.; Chen, C.; Wang, K. Li e Combined B. sub ilis and E. aecium Alle ia e Li e In lamma ion, Imp o e
In es inal Ba ie Func ion, and Modula e Gu Mic obio a in Mice wi h Non-Alcoholic Fa y Li e Disease. Med. Sci. Moni .
2021
,
27, e931143. [C ossRe ] [PubMed]
49.
Plaza-Diaz, J.; Gomez-Llo en e, C.; Abadia-Molina, F.; Saez-La a, M.J.; Campaña-Ma in, L.; Muñoz-Quezada, S.; Rome o, F.;
Gil, A.; Fon ana, L. E ec s o Lac obacillus pa acasei CNCM I-4034, Bi idobac e ium b e e CNCM I-4035 and Lac obacillus hamnosus
CNCM I-4036 on Hepa ic S ea osis in Zucke Ra s. PLoS ONE 2014,9, e98401. [C ossRe ]
50. Al-Muza a , H.M.; Amin, K.A. P obio ic mix u e imp o es a y li e disease by i ue o i s ac ion on lipid p o iles, lep in, and
in lamma o y bioma ke s. BMC Complemen . Al e n. Med. 2017,17, 43. [C ossRe ]
51.
Kim, S.-J.; Pa k, S.H.; Sin, H.-S.; Jang, S.-H.; Lee, S.-W.; Kim, S.-Y.; Kwon, B.; Yu, K.-Y.; Kim, S.Y.; Yang, D.K. Hypocholes e olemic
e ec s o p obio ic mix u e on die -induced hype choles e olemic a s. Nu ien s 2017,9, 293. [C ossRe ]
52.
Candelli, M.; F anza, L.; Pigna a o, G.; Oje i, V.; Co ino, M.; Piccioni, A.; Gasba ini, A.; F anceschi, F. In e ac ion be ween
Lipopolysaccha ide and Gu Mic obio a in In lamma o y Bowel Diseases. In . J. Mol. Sci. 2021,22, 6242. [C ossRe ] [PubMed]
53.
Alle , R.; De Luis, D.A.; Izaola, O.; Conde, R.; Gonzalez Sag ado, M.; P imo, D.; De La Fuen e, B.; Gonzalez, J. E ec o a p obio ic
on li e amino ans e ases in nonalcoholic a y li e disease pa ien s: A double blind andomized clinical ial. Eu . Re . Med.
Pha macol. Sci. 2011,15, 1090–1095. [PubMed]
54.
Famou i, F.; Sha ia , Z.; Hashemipou , M.; Keikha, M.; Kelishadi, R. E ec s o P obio ics on Nonalcoholic Fa y Li e Disease in
Obese Child en and Adolescen s. J. Pedia . Gas oen e ol. Nu . 2017,64, 413–417. [C ossRe ] [PubMed]
55.
Kobyliak, N.; Abena oli, L.; Mykhalchyshyn, G.; Kononenko, L.; Boccu o, L.; Ky iienko, D.; Dynnyk, O. A Mul i-s ain P obio ic
Reduces he Fa y Li e Index, Cy okines and Amino ans e ase le els in NAFLD Pa ien s: E idence om a Randomized Clinical
T ial. J. Gas oin es . Li e Dis. 2018,27, 41–49. [C ossRe ]
56.
Ahn, S.B.; Jun, D.W.; Kang, B.; Lim, J.H.; Lim, S.; Chung, M.J. Randomized, Double-blind, Placebo-con olled S udy o a
Mul ispecies P obio ic Mix u e in Nonalcoholic Fa y Li e Disease. Sci. Rep. 2019,9, 5688. [C ossRe ]
57.
Duseja, A.; Acha ya, S.K.; Meh a, M.; Chhab a, S.; Rana, S.; Das, A.; Da agup a, S.; Dhiman, R.K.; Chawla, Y.K. High po ency
mul is ain p obio ic imp o es li e his ology in non-alcoholic a y li e disease (NAFLD): A andomised, double-blind, p oo o
concep s udy. BMJ Open Gas oen e ol. 2019,6, e000315. [C ossRe ]
58.
Beh ouz, V.; A yaeian, N.; Zahedi, M.J.; Jazaye i, S. E ec s o p obio ic and p ebio ic supplemen a ion on me abolic pa ame e s,
li e amino ans e ases, and sys emic in lamma ion in nonalcoholic a y li e disease: A andomized clinical ial. J. Food Sci.
2020,85, 3611–3617. [C ossRe ]
In . J. Mol. Sci. 2022,23, 3167 24 o 25
59.
Naba i, S.; Ra a , M.; Somi, M.H.; Homayouni-Rad, A.; Asgha i-Ja a abadi, M. E ec s o p obio ic yogu consump ion on
me abolic ac o s in indi iduals wi h nonalcoholic a y li e disease. J. Dai y Sci. 2014,97, 7386–7393. [C ossRe ]
60.
Chong, P.L.; Laigh , D.; Aspinall, R.J.; Higginson, A.; Cummings, M.H. A andomised placebo con olled ial o VSL#3
®
p obio ic
on bioma ke s o ca dio ascula isk and li e inju y in non-alcoholic a y li e disease. BMC Gas oen e ol. 2021,21, 144.
61.
Mohamad No , M.H.; Ayob, N.; Mokh a , N.M.; Raja Ali, R.A.; Tan, G.C.; Wong, Z.; Sha iee, N.H.; Wong, Y.P.; Mus angin,
M.; Nawawi, K.N.M. The E ec o P obio ics (MCP
®
BCMC
®
S ains) on Hepa ic S ea osis, Small In es inal Mucosal Immune
Func ion, and In es inal Ba ie in Pa ien s wi h Non-Alcoholic Fa y Li e Disease. Nu ien s 2021,13, 3192. [C ossRe ]
62. Xu, C.; Zhu, H.; Qiu, P. Aging p og ession o human gu mic obio a. BMC Mic obiol. 2019,19, 236. [C ossRe ] [PubMed]
63.
Han, A.; Zhang, Y.N.; Boeh inge , A.S.; Mon es, V.; And e, M.P.; E dman, J.W., J .; Loomba, R.; Valasek, M.A.; Si lin, C.B.;
O’B ien, W.D., J . Assessmen o Hepa ic S ea osis in Nonalcoholic Fa y Li e Disease by Using Quan i a i e US. Radiology
2020
,
295, 106–113. [C ossRe ] [PubMed]
64.
Yada , H.; Lee, J.; Lloyd, J.; Wal e , P.; Rane, S.G. Bene icial Me abolic E ec s o a P obio ic ia Bu y a e-induced GLP-1 Ho mone
Sec e ion. J. Biol. Chem. 2013,288, 25088–25097. [C ossRe ]
65.
E e a d, A.; Belze , C.; Geu s, L.; Ouwe ke k, J.P.; D ua , C.; Bindels, L.B.; Guio , Y.; De ien, M.; Muccioli, G.G.;
Delzenne, N.M.; e al
. C oss- alk be ween Akke mansia muciniphila and in es inal epi helium con ols die -induced obesi y. P oc.
Na l. Acad. Sci. USA 2013,110, 9066–9071. [C ossRe ]
66.
Minami, J.; Kondo, S.; Yanagisawa, N.; Odamaki, T.; Xiao, J.; Abe, F.; Nakajima, S.; Hamamo o, Y.; Sai oh, S.; Shimoda, T.
O al adminis a ion o Bi idobac e ium b e e B-3 modi ies me abolic unc ions in adul s wi h obese endencies in a andomised
con olled ial. J. Nu . Sci. 2015,4, e17. [C ossRe ] [PubMed]
67.
Fi ouzi, S.; Majid, H.A.; Ismail, A.; Kama uddin, N.A.; Ba aka un-Nisak, M.Y. E ec o mul i-s ain p obio ics (mul i-s ain
mic obial cell p epa a ion) on glycemic con ol and o he diabe es- ela ed ou comes in people wi h ype 2 diabe es: A andomized
con olled ial. Eu . J. Nu . 2017,56, 1535–1550. [C ossRe ]
68.
Asad, F.; Anwa , H.; Yassine, H.M.; Ullah, M.I.; Rahman, A.; Kam an, Z.; Sohail, M.U. Whi e Bu on Mush oom, Aga icus bispo us
(Aga icomyce es), and a P obio ics Mix u e Supplemen a ion Co ec Dyslipidemia wi hou In luencing he Colon Mic obiome
P o ile in Hype choles e olemic Ra s. In . J. Med. Mush ooms 2020,22, 235–244. [C ossRe ]
69.
Yao, M.; Q , L.; Lu, Y.; Wang, B.; Be glund, B.; Li, L. An Upda e on he E icacy and Func ionali y o P obio ics o he T ea men
o Non-Alcoholic Fa y Li e Disease. Enginee ing 2021,7, 679–686. [C ossRe ]
70.
E xebe ia, U.; Hijona, E.; Agui e, L.; Milag o, F.I.; Bujanda, L.; Rimando, A.M.; Ma ínez, J.A.; Po illo, M.P. P e os ilbene-induced
changes in gu mic obio a composi ion in ela ion o obesi y. Mol. Nu . Food Res. 2017,61, 1500906. [C ossRe ]
71.
Mo ei a, G.V.; Aze edo, F.F.; Ribei o, L.M.; San os, A.; Guadagnini, D.; Gama, P.; Libe i, E.A.; Saad, M.J.A.; Ca alho, C.R.O.
Li aglu ide modula es gu mic obio a and educes NAFLD in obese mice. J. Nu . Biochem. 2018,62, 143–154. [C ossRe ]
72.
Cani, P.D.; de Vos, W.M. Nex -Gene a ion Bene icial Mic obes: The Case o Akke mansia muciniphila.F on . Mic obiol.
2017
,8, 1765.
[C ossRe ] [PubMed]
73.
Seo, B.; Jeon, K.; Moon, S.; Lee, K.; Kim, W.K.; Jeong, H.; Cha, K.H.; Lim, M.Y.; Kang, W.; Kweon, M.N.; e al. Rosebu ia spp.
Abundance Associa es wi h Alcohol Consump ion in Humans and I s Adminis a ion Amelio a es Alcoholic Fa y Li e in Mice.
Cell Hos Mic obe 2020,27, 25–40.e6. [C ossRe ] [PubMed]
74. Do on, S.; Snydman, D.R. Risk and Sa e y o P obio ics. Clin. In ec . Dis. 2015,60, S129–S134. [C ossRe ]
75.
Cue as-González, P.F.; Liceaga, A.M.; Aguila -Toalá, J.E. Pos bio ics and pa ap obio ics: F om concep s o applica ions. Food Res.
In . 2020,136, 109502. [C ossRe ]
76.
Na a aj, B.H.; Ali, S.A.; Beha e, P.V.; Yada , H. Pos bio ics-pa abio ics: The new ho izons in mic obial bio he apy and unc ional
oods. Mic ob. Cell Fac . 2020,19, 168. [C ossRe ] [PubMed]
77.
Vallejo-Co doba, B.; Cas o-López, C.; Ga cía, H.S.; González-Có do a, A.F.; He nández-Mendoza, A. Pos bio ics and pa ap obi-
o ics: A e iew o cu en e idence and eme ging ends. Ad . Food Nu . Res. 2020,94, 1–34.
78.
Piqué, N.; Be langa, M.; Miñana-Galbis, D. Heal h Bene i s o Hea -Killed (Tyndallized) P obio ics: An O e iew. In . J. Mol. Sci.
2019,20, 2534. [C ossRe ]
79.
Kang, X.; Liang, H.; Luo, Y.; Li, Z.; He, F.; Han, X.; Zhang, L. An i-adipogenesis and me abolism- egula ing e ec s o hea -
inac i a ed S ep ococcus he mophilus MN-ZLW-002. Le . Appl. Mic obiol. 2020,72, 677–687. [C ossRe ]
80.
Nakamu a, F.; Ishida, Y.; Aiha a, K.; Sawada, D.; Ashida, N.; Sugawa a, T.; Aoki, Y.; Takeha a, I.; Takano, K.; Fujiwa a, S. E ec o
agmen edLac obacillus amylo o usCP1563 on lipid me abolism in o e weigh and mildly obese indi iduals: A andomized
con olled ial. Mic ob. Ecol. Heal h Dis. 2016,27, 30312. [C ossRe ]
81.
Zo zela, L.; A des ani, S.K.; Mc a land, L.V.; Voh a, S. Is he e a ole o modi ied p obio ics as bene icial mic obes: A sys ema ic
e iew o he li e a u e. Bene . Mic obes 2017,8, 739–754. [C ossRe ]
82.
Peluzio, M.D.C.G.; Ma inez, J.A.; Milag o, F.I. Pos bio ics: Me aboli es and mechanisms in ol ed in mic obio a-hos in e ac ions.
T ends Food Sci. Technol. 2021,108, 11–26. [C ossRe ]
83.
Wang, S.; Ahmadi, S.; Nagpal, R.; Jain, S.; Mish a, S.P.; Ka anagh, K.; Zhu, X.; Wang, Z.; McClain, D.A.; K i che sky, S.B.; e al.
Lipo eichoic acid om he cell wall o a hea killed Lac obacillus pa acasei D3-5 amelio a es aging- ela ed leaky gu , in lamma ion
and imp o es physical and cogni i e unc ions: F om C. elegans o mice. Ge oScience 2020,42, 333–352. [C ossRe ] [PubMed]
In . J. Mol. Sci. 2022,23, 3167 25 o 25
84.
Ma, L.; Ni, Y.; Wang, Z.; Tu, W.; Ni, L.; Zhuge, F.; Zheng, A.; Hu, L.; Zhao, Y.; Zheng, L.; e al. Spe midine imp o es gu ba ie
in eg i y and gu mic obio a unc ion in die -induced obese mice. Gu Mic obes 2020,12, 1832857. [C ossRe ] [PubMed]
85.
Ma, L.; Ni, Y.; Hu, L.; Zhao, Y.; Zheng, L.; Yang, S.; Ni, L.; Fu, Z. Spe midine amelio a es high- a die -induced hepa ic s ea osis
and adipose issue in lamma ion in p eexis ing obese mice. Li e Sci. 2021,265, 118739. [C ossRe ] [PubMed]
