Co esponding au ho : Basiouny Fouad El-sendiony
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P o ec i e Roles o L-ca ni ine and Ginkgo biloba Ex ac on Oxida i e S ess and
Neu o ansmi e Imbalance in Pen ylene e azole-Induced Epilepsy in Ra s
Basiouny Fouad El-sendiony * and Abdulaziz M. Hakami
Physiology Depa men , AlGhad College o Applied Medical Sciences, Naj an 66243, Saudi A abia.
GSC Biological and Pha maceu ical Sciences, 2025, 33(01), 129-133
Publica ion his o y: Recei ed on 04 Sep embe 2025; e ised on 12 Oc obe 2025; accep ed on 14 Oc obe 2025
A icle DOI: h ps://doi.o g/10.30574/gscbps.2025.33.1.0395
Abs ac
Epilepsy is a neu ological diso de ma ked by oxida i e s ess and neu o ansmi e imbalance, d i ing neu onal
damage and seizu es. This s udy e alua ed biochemical and neu ochemical changes in pen ylene e azole (PTZ)-
kindled seizu es and he p o ec i e e ec s o L-ca ni ine and Ginkgo biloba ex ac (GBE). Eigh y male albino a s we e
di ided in o con ol, L-ca ni ine, GBE, PTZ, and PTZ wi h p e- o pos - ea men o L-ca ni ine o GBE. Epilepsy was
induced ia PTZ injec ions (40 mg/kg) e e y 48 hou s o h ee weeks. Oxida i e s ess ma ke s—MDA, GSH, GPx, SOD,
and CAT—and neu o ansmi e s—GABA, glu ama e, and AChE—we e measu ed in se um and ce eb al co ex. PTZ
igge ed oxida i e s ess and exci a o y imbalance, shown by inc eased MDA, glu ama e, and AChE, alongside educed
an ioxidan s and GABA. P e ea men wi h L-ca ni ine o GBE signi ican ly es o ed an ioxidan le els and
neu o ansmi e balance; pos - ea men achie ed mode a e imp o emen . L-ca ni ine had s onge an ioxidan
e ec s, while GBE mo e e ec i ely no malized neu o ansmi e s. These esul s sugges bo h agen s o e
neu op o ec ion by educing oxida i e s ess and es o ing inhibi o y–exci a o y balance, suppo ing hei po en ial as
adjunc he apies in epilepsy.
Keywo ds: Epilepsy; Pen ylene e azole; Oxida i e S ess; Neu o ansmi e s; L-Ca ni ine; Ginkgo Biloba;
An ioxidan s
1. In oduc ion
Epilepsy is one o he mos common ch onic neu ological diseases, a ec ing oughly 1 % o he global popula ion. I is
cha ac e ized by ecu en unp o oked seizu es ha e lec excessi e and synch onized neu onal i ing. Al hough
se e al mechanisms con ibu e o epilep ogenesis, accumula ing e idence implica es oxida i e s ess and
neu o ansmi e imbalance as c i ical pa hophysiological ac o s [1; 2].
Seizu e ac i i y leads o inc eased oxygen consump ion and mi ochond ial o e load, p oducing la ge amoun s o
eac i e oxygen species (ROS). These ROS a ack memb ane lipids, p o eins, and DNA, ini ia ing neu onal degene a ion.
Meanwhile, dis up ion o neu o ansmi e homeos asis pa icula ly a dec ease in he inhibi o y ansmi e γ-
aminobu y ic acid (GABA) and an inc ease in he exci a o y amino acid glu ama e c ea es a hype exci able neu al
en i onmen ha p omo es seizu e ecu ence [3; 4].
Pen ylene e azole (PTZ) is a con ulsan ha induces seizu es by an agonizing he GABA_A ecep o , he eby educing
inhibi o y neu o ansmission. Ch onic adminis a ion o PTZ p oduces a eliable animal model o gene alized epilepsy
wi h accompanying oxida i e s ess, making i sui able o e alua ing neu op o ec i e agen s [5].
L-ca ni ine, a qua e na y ammonium compound syn hesized om lysine and me hionine, acili a es mi ochond ial β-
oxida ion o a y acids and ene gy p oduc ion. I exhibi s no able an ioxidan and an i-apop o ic e ec s [6; 7]. Ginkgo
GSC Biological and Pha maceu ical Sciences, 2025, 33(01), 129-133
130
biloba ex ac (GBE), de i ed om Ginkgo biloba lea es, con ains la onoids and e penoids ha possess s ong ee-
adical-sca enging, memb ane-s abilizing, and an i-in lamma o y p ope ies [8].
Despi e he indi idual neu op o ec i e e idence o L-ca ni ine and GBE, compa a i e and combined analyses a ge ing
oxida i e and neu ochemical pa ame e s in PTZ-induced epilepsy emain limi ed.
This s udy he e o e aimed o
• Cha ac e ize oxida i e and neu o ansmi e al e a ions in he PTZ-kindling model; and
• E alua e he amelio a i e e ec s o L-ca ni ine and GBE, adminis e ed bo h be o e and a e seizu e induc ion,
on hese biochemical and neu ochemical dis u bances.
2. Ma e ials and Me hods
2.1. Animals
Eigh y adul male albino a s (Ra us no egicus, 7–8 weeks old, 120 ± 10 g) we e housed unde con olled empe a u e
(23 ± 2 °C), humidi y (55 ± 5 %), and 12 h ligh /da k cycle. They ecei ed s anda d chow and wa e ad libi um. All
p ocedu es complied wi h ins i u ional e hical guidelines o animal expe imen a ion.
2.2. Expe imen al Design
Animals we e andomly assigned o eigh g oups (n = 10)
Table 1: Expe imen al g ouping and ea men p o ocols o animal s udy.
G oup
T ea men
G1
Con ol (saline)
G2
L-ca ni ine (300 mg/kg i.p. e e y 48 h × 3 weeks)
G3
GBE (100 mg/kg i.p. e e y 48 h × 3 weeks)
G4
PTZ (40 mg/kg i.p. e e y 48 h × 3 weeks)
G5
L-ca ni ine p e- ea ed + PTZ
G6
GBE p e- ea ed + PTZ
G7
PTZ + L-ca ni ine pos - ea ed
G8
PTZ + GBE pos - ea ed
2.3. Induc ion o Epilepsy
PTZ (Sigma-Ald ich) was adminis e ed in ape i oneally a 40 mg/kg e e y 48 h o 21 days. Beha io al seizu es we e
obse ed and sco ed acco ding o Racine’s scale (0–5). Consis en s age 4–5 seizu es con i med success ul kindling [9].
2.4. Sample Collec ion
Twen y- ou hou s a e he inal injec ion, a s we e anes he ized and decapi a ed. Blood was collec ed o se um
sepa a ion; b ains we e apidly excised, and ce eb al co ices we e dissec ed, weighed, and homogenized in cold
phospha e bu e (0.1 M, pH 7.4). The supe na an s we e used o biochemical assays.
2.5. Biochemical and Neu ochemical Assays
• Lipid pe oxida ion (Malondialdhyde; MDA) – hioba bi u ic acid- eac i e subs ances (TBARS) me hod.
• Reduced glu a hione (GSH) – Ellman’s eagen (5,5′-di hiobis-2-ni obenzoic acid).
• An ioxidan enzymes: glu a hione pe oxidase (GPx), supe oxide dismu ase (SOD), and ca alase (CAT) we e
assayed using colo ime ic ki s (Biodiagnos ic, Egyp ).
• Neu o ansmi e s: GABA and glu ama e we e quan i ied by HPLC wi h p e-column de i a iza ion using o-
ph halaldehyde.
• Ace ylcholines e ase (AChE) ac i i y was measu ed spec opho ome ically ollowing Ellman’s me hod.
GSC Biological and Pha maceu ical Sciences, 2025, 33(01), 129-133
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2.6. S a is ical Analysis
Da a we e exp essed as mean ± SEM. S a is ical di e ences we e analyzed by one-way ANOVA (p ≤ 0.05) using SPSS
25.
3. Resul s
3.1. Gene al Obse a ions
Con ol, L-ca ni ine, and GBE-only g oups exhibi ed no mal beha io and weigh gain. PTZ- ea ed a s displayed
p og essi e seizu e ac i i y wi h onic–clonic con ulsions and mild weigh loss. P e ea men wi h ei he compound
delayed seizu e onse and educed se e i y sco es.
3.2. Oxida i e S ess Bioma ke s
PTZ-kindled a s showed a ma ked ise in MDA le els accompanied by signi ican educ ions in GSH, GPx, SOD, and CAT
ac i i ies, con i ming oxida i e s ess. L-ca ni ine and GBE p e ea men s signi ican ly lowe ed MDA and es o ed
an ioxidan enzyme ac i i ies owa d no mal. Pos - ea men achie ed pa ial eco e y.
Table 2 Se um and Co ical An ioxidan Ma ke s (Mean ± SEM)
Pa ame e
Con ol
PTZ
P e + LC
P e + GBE
Pos + LC
Pos + GBE
MDA (nmol/mg p o ein)
1.2 ± 0.1
3.8 ± 0.3 ↑↑
1.6 ± 0.2
1.8 ± 0.2
2.0 ± 0.2
2.1 ± 0.3
GSH (µmol/mg)
8.4 ± 0.4
4.1 ± 0.3 ↓↓
7.8 ± 0.3
7.5 ± 0.4
6.9 ± 0.4
6.7 ± 0.3
GPx (U/mg)
25 ± 2
13 ± 1 ↓↓
22 ± 2
20 ± 2
18 ± 1
17 ± 1
SOD (U/mg)
15 ± 1
8 ± 0.5 ↓↓
14 ± 0.7
13 ± 0.6
12 ± 0.6
11 ± 0.5
CAT (U/mg)
50 ± 4
27 ± 3 ↓↓
46 ± 3
44 ± 3
40 ± 3
39 ± 2
3.3. Neu o ansmi e and AChE Al e a ions
PTZ signi ican ly dec eased GABA (–45 %) and inc eased glu ama e (+70 %) and AChE ac i i y (+60 %) ela i e o
con ols. L-ca ni ine and GBE e e sed hese changes, pa icula ly when gi en as p e ea men s.
Table 3 Co ical Neu o ansmi e s and AChE Ac i i y
Pa ame e
Con ol
PTZ
P e + LC
P e + GBE
Pos + LC
Pos + GBE
GABA (µmol/g issue)
4.6 ± 0.3
2.5 ± 0.2 ↓↓
4.3 ± 0.3
4.1 ± 0.3
3.7 ± 0.2
3.5 ± 0.3
Glu ama e (µmol/g)
3.2 ± 0.2
5.5 ± 0.4 ↑↑
3.4 ± 0.3
3.6 ± 0.3
3.9 ± 0.3
4.0 ± 0.3
AChE (U/mg p o ein)
0.45 ± 0.03
0.72 ± 0.05 ↑
0.48 ± 0.04
0.50 ± 0.04
0.53 ± 0.05
0.55 ± 0.05
3.4. Compa a i e E icacy
L-ca ni ine p e ea men no malized an ioxidan enzymes mo e e icien ly han GBE, whe eas GBE p oduced a mo e
p onounced inc ease in GABA and dec ease in AChE ac i i y. Combined da a sugges complemen a y mechanisms o
neu op o ec ion.
4. Discussion
4.1. Oxida i e S ess in Epilepsy
PTZ-kindled seizu es elici ed se e e oxida i e imbalance, e lec ed by ele a ed MDA and deple ion o GSH and ela ed
enzymes. The high me abolic demand du ing epe i i e seizu es accele a es mi ochond ial elec on leakage and ROS
gene a ion. Lipid pe oxida ion o neu onal memb anes impai s ion channels and neu o ansmi e elease, agg a a ing
exci o oxici y. Simila oxida i e signa u es ha e been epo ed in human epilep ic issues and PTZ-kindled a s [2; 10].
GSC Biological and Pha maceu ical Sciences, 2025, 33(01), 129-133
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4.2. An ioxidan Res o a ion by L-ca ni ine and GBE
L-ca ni ine’s abili y o shu le long-chain a y acids in o mi ochond ia enhances β-oxida ion e iciency, educing
elec on leakage and ROS o ma ion. I s hiol g oup and me al-chela ing capaci y con e di ec an ioxidan e ec s.
Res o a ion o GSH and CAT obse ed he e aligns wi h p e ious epo s showing L-ca ni ine’s capaci y o up egula e
an ioxidan gene exp ession and p o ec agains neu onal apop osis [11].
GBE exe s i s an ioxidan e ec p ima ily h ough la onoids such as que ce in and kaemp e ol, which sca enge
hyd oxyl and supe oxide adicals. Te penoids (ginkgolides, bilobalide) also inhibi lipid pe oxida ion and s abilize
mi ochond ial memb anes. The pa ial no maliza ion o oxida i e pa ame e s in GBE- ea ed a s indica es subs an ial
sys emic an ioxidan ac ion [12].
4.3. Neu o ansmi e Modula ion
PTZ-induced supp ession o GABA and ele a ion o glu ama e con i m an exci a o y/inhibi o y imbalance—a hallma k
o epilepsy. Res o a ion o GABA and educ ion o glu ama e by bo h agen s sugges enhancemen o inhibi o y
neu o ansmission and a enua ion o exci o oxic signaling. L-ca ni ine may modula e GABAe gic one indi ec ly ia
imp o ed mi ochond ial ATP gene a ion equi ed o GABA syn hesis. GBE may ac h ough modula ion o NMDA-
ecep o ac i i y and up- egula ion o glu amic acid deca boxylase (GAD), he enzyme ha con e s glu ama e o GABA
[13].
4.4. Ace ylcholines e ase Ac i i y
Ele a ed AChE in PTZ a s e lec s choline gic hype ac i i y and oxida i e modi ica ion o he enzyme. T ea men wi h
ei he compound signi ican ly dec eased AChE ac i i y, sugges ing imp o ed choline gic balance and educed neu onal
s ess. GBE’s la onoids a e known AChE inhibi o s, which may enhance cogni i e esilience and synap ic s abili y [14].
4.5. Compa a i e and Syne gis ic Aspec s
While bo h agen s ac as an ioxidan s, hei modes di e : L-ca ni ine enhances ene gy me abolism, whe eas GBE
p o ides memb ane-le el an ioxidan and ecep o modula ion. P e ea men a o ded be e ou comes han pos -
ea men , emphasizing he impo ance o p ophylac ic an ioxidan suppo in seizu e managemen .
4.6. Mechanis ic In eg a ion
The neu op o ec i e ou come can be a ibu ed o mul iple syne gis ic mechanisms
• Supp ession o ROS gene a ion and lipid pe oxida ion;
• Enhancemen o endogenous an ioxidan enzyme syn hesis;
• Res o a ion o GABA/glu ama e equilib ium;
• Regula ion o choline gic ac i i y ia AChE inhibi ion; and
• S abiliza ion o neu onal memb anes and mi ochond ia.
By a enua ing oxida i e inju y and neu o ansmi e dys egula ion, L-ca ni ine and GBE p e en he cascade leading
om oxida i e s ess o neu onal dea h.
4.7. Clinical Rele ance
Ch onic an iepilep ic d ug use o en exace ba es oxida i e s ess. Na u al an ioxidan s wi h neu ochemical s abilizing
p ope ies could he e o e se e as adju an he apies o imp o e seizu e con ol and mi iga e sys emic side e ec s.
The cu en esul s suppo u u e ansla ional s udies and possible combina ion he apies.
5. Conclusion
Pen ylene e azole-induced epilepsy p o okes signi ican oxida i e s ess and neu o ansmi e imbalance in a
ce eb al co ex. L-ca ni ine and Ginkgo biloba ex ac e ec i ely coun e hese pa hological changes h ough po en
an ioxidan and neu omodula o y mechanisms. P e ea men wi h ei he compound p o ides subs an ial p o ec ion,
whe eas pos - ea men aids eco e y. Thei complemen a y ac ions—me abolic suppo by L-ca ni ine and ecep o -
le el an ioxidan de ense by GBE—highligh hei p omise as adjunc i e, non- oxic s a egies o p e en ing oxida i e
neu onal damage in epilep ic condi ions.
GSC Biological and Pha maceu ical Sciences, 2025, 33(01), 129-133
133
Compliance wi h e hical s anda ds
Acknowledgmen s
To all pa icipan s in he s udy. To D . Basiouny Fouad El-sendiony o his suppo in all he ime o esea ch. To D .
Abdulaziz M. Hakami con inuous encou agemen h oughou his wo k.
Disclosu e o con lic o in e es
The au ho s decla e ha hey ha e no con lic o in e es o disclose.
S a emen o e hical app o al
Animals used o esea ch only wi hin an e hical amewo k.
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