Co esponding au ho : Jude Nnaemeka Okoyeh
Copy igh © 2025 Au ho (s) e ain he copy igh o his a icle. This a icle is published unde he e ms o he C ea i e Commons A ibu ion Liscense 4.0.
Hepa op o ec i e E ec s o Vi ex Doniana F ui Ex ac on Ca bon Te achlo ide
Induced Li e Damage
Vic o ia Nonyelum Olli 1, Jude Nnaemeka Okoyeh 2, * and Kosisochukwu Chizoba Udechukwu 3
1 Depa men o Pha macology and Toxicology, Chukwuemeka Odumegwu Ojukwu Uni e si y Igba iam campus Nige ia.
2 Depa men o Medical Labo a o y Science and Biology, School o Nu sing and Heal h Sciences, Neumann Uni e si y, One
Neumann D i e, As on, PA, 19014. USA.
3 Depa men o Pha macology and Toxicology, Chukwuemeka Odumegwu Ojukwu Uni e si y Igba iam campus Nige ia.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(02), 989-996
Publica ion his o y: Recei ed on 05 July 2025; e ised on 10 Augus ; accep ed on 13 Augus 2025
A icle DOI: h ps://doi.o g/10.30574/wja .2025.27.2.2934
Abs ac
The li e in he cou se o playing i s ole o de oxi ica ion, p oduces highly eac i e molecules known as ee adicals
which can damage he li e . Hence, in spi e o he ad ancemen in he ield o hepa hology, li e disease is s ill on he
inc ease. The need o a na u al hepa op o ec i e agen mo i a ed his s udy. Acu e oxici y s udy and phy ochemical
sc eening o he e hanol ui ex ac o Vi ex doniana was done. The hepa op o ec i e s udy on he ex ac was ca ied
ou using adul albino a s. The li e damage was induced by he subcu aneous adminis a ion o ca bon e achlo ide
(CCl4) in a single dose o 2 ml/kgbw pe day o 4 days. The a s we e hen as ed o 36 h be o e he o al adminis a ion
o he ex ac s (250, 500 and 1000 mg/kg) o 21 days. E ec o he ex ac on his opa hology o he li e was also
in es iga ed.
A signi ican dose-dependen (P<0.05) educ ion in he le els o ALT, AST and ALP enzymes was obse ed in he
ea men g oups when compa ed o he nega i e con ol. The ex ac signi ican ly (P<0.05) inc eased he se um o al
p o ein when compa ed o nega i e con ol. Fu he mo e, a dose-dependen es o a ion o he li e a 100, 250, 500
and 1000 mg/kg dose o he ui ex ac was obse ed. Conside ing he da a ob ained om his s udy, he ui ex ac
o Vi ex doniana could possesse hypa op o ec i e p ope y.
Keywo ds: Vi ex doniana; Lli e damage; Hepa op o ec i e; Ca bon e achlo ide
1. In oduc ion
The ques o e icacious emedies o ea a ious human ailmen s has cen e ed on he u iliza ion o medicinal plan s
[1]. As heal h ca e cos s con inue o escala e, he a ac ion o low-cos medicinal emedies has s imula ed he in e es
o consume s o e-e alua e he po en ial o al e na i es [2]. Some he bs ha e been employed in he ea men o li e
ailmen s [3]. The li e is a i al o gan ha pe o ms nume ous unc ions essen ial o me abolism, immuni y, diges ion,
de oxi ica ion, and i amin s o age [4]
Ca bon e achlo ide (CCl4) is a well-known hepa o oxin capable o causing li e damage [5]. D ug-induced li e inju y
accoun s o app oxima ely i y pe cen o cases o acu e li e ailu e globally [6], and app oxima ely 29 million people
wo ldwide su e om ch onic li e disease [7]. Some o he causes o li e disease wo ldwide a e ch onic hepa i is B
and C, alcohol and non-alcoholic s ea ohepa i is associa ed wi h obesi y and me abolic synd ome [8].
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Dis up ion o he li e unc ions esul s in he excessi e elease o oxidan s ha damage hepa ic cells. Addi ionally,
ac i a ion o speci ic enzymes wi hin he cy och ome P-450 sys em, pa icula ly CYP2E1, con ibu es o oxida i e s ess
[9]. Se e al li e unc ion es s a e a ailable and, in mos cases, he ea ly signs o li e inju y is ele a ion o hepa ic
enzymes.
Classes o d ugs used in li e oxici y ea men include immunosupp essan s[10], glucoco icoids[11], combina ion o
he apeu ic d ugs[12], N-ace ylcys eine [13] and glu a hione [14].
Vi ex doniana ee is a pe ennial ee o en g own o i s ui s. I is widesp ead in opical A ica and is commonly
known as Black plum o A ican oli e, Dinya (Hausa), Galbihi (Fulani), Oo i-nla (Yo uba), Uchaco o/mbe mbe (Igbo). I
is widesp ead in he sou hwes e n and sou heas e n Nige ia [15] wi h mul iple uses such as ood (bo h lea es and
ui s) and medicines. I is commonly used in adi ional se ings o he managemen o a numbe o diseases; a
decoc ion o he woody pa s o he plan is used o he ea men o in lamma o y diso de s, s omach pains, dia hea,
heuma ic pains and dysen e y [16]. The young lea es and oo s a e used o he ea men o wounds and male
impo ence espec i ely, and he ui ex ac is used in he ea men o a igue and cons ipa ion [17].
Figu e 1 The ui s o Vi ex doniana
2. Ma e ial and me hods
2.1. Ma e ials
2.1.1. Plan ma e ials
Sample collec ion and Iden i ica ion: The esh ui s o Vi ex doniana we e pu chased om Ekwulobia in Anamb a
s a e o Nige ia. The ui was au hen ica ed by a Taxonomis a he depa men o Bo any, Nnamdi Azikiwe Uni e si y,
Awka, Nige ia, and a ouche specimen numbe (NAUBT 3218) was assigned o i .
2.1.2. Ex ac ion o plan ma e ial
Ex ac ion was ca ied ou using a me hod ha was p e iously desc ibed [18]. The ui s we e cleaned o emo e sand
and o he deb is. A e emo ing he hin me ica p, he leshy juicy mesoca p was sc aped o om he seeds and d ied
o a pe iod o wo weeks unde oom empe a u e. This d ied mesoca p was g inded in o powde wi h he help o an
elec ical g inde . Abou 800 g o he powde ed ma e ial was cold mace a ed in 80 % e hanol. The mix u e was agi a ed
in e mi en ly o h ee days (72 hou s) be o e il a ion. The il a e was eco e ed and concen a ed o d yness using
wa e ba h a 40 OC. The pe cen age yield o he ex ac was calcula ed and he ex ac s o ed in a e ige a o un il
when needed o expe imen s.
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2.1.3. Animals
Thi y- i e albino a s o bo h sexes weighing 130g-150g, pu chased om he labo a o y animal house o Chukwuemeka
Odumegwu Ojukwu Uni e si y, Igba iam, Nige ia, we e used o he s udy. E hical app o al numbe
PHACOOU/AREC/2023/032 was assigned o a es ha he animals we e ca ed o acco ding o he Facul y o Pha macy
(COOU) Animal Resea ch E hics Commi ee Guidelines (PHACOOUAREC), which a e in con o mi ies wi h he Na ional
Ins i u e o Heal h (NIH), USA, guidelines o he ca e and use o labo a o y animals.
2.2. Me hods
2.2.1. Quali a i e phy ochemical analysis:
Sc eening o he p esence o seconda y me aboli es was ca ied ou ollowing he phy ochemical es s as demons a ed
ea lie [19]. The p esence o he ollowing seconda y me aboli es we e es ed o : annins, alkaloids, educing suga s,
la onoids, glycosides, an h aquinones, saponins, acidic compounds and p o eins.
2.2.2. Acu e oxici y s udy:
The acu e oxici y es was conduc ed using he up and down p ocedu e (UDP) adop ed [20] and e ised [21]. Using his
me hod, he animals we e dosed one a a ime and he doses we e dependen on he esponse o he i s animal o he
ini ial dose. The second animal ecei es a lowe dose i he i s animal dies ( he ini ial dose is dec eased by a ac o o
3.2) o he second animal ecei es a highe dose i he i s animal su i es ( he ini ial dose is inc eased by a ac o o
3.2). Th ee albino a s (130-150 g) we e used as s a ing poin . Two a s se ed as nega i e con ol ha ing ecei ed 10
ml/kg o dis illed wa e o ally while he es animal ecei ed a de aul o al dose o 5000 mg/kg o he ex ac . The
animals we e hen obse ed con inuously o 4 hou s o changes in beha io and o any o he ob ious signs o oxici y,
and subsequen ly daily o addi ional 14 days o delayed oxici y.
2.2.3. Induc ion o li e damage
The li e damage was induced by he subcu aneous adminis a ion o ca bon e achlo ide (CCl4) in oli e oil (50% /
CCl4) in a single dose o 2 ml/kg body weigh pe day o 4 days. The ea e , he a s we e as ed o 36 h be o e he
adminis a ion o he ex ac s.
2.2.4. Animal g ouping and ea men
The a s we e andomly di ided in o 7 g oups (o 5 pe g oup) as ollows:
• G oup 1: No mal con ol ( ecei ed eed and wa e only)
• G oup 2: T ea ed wi h oli e oil and se ed as ehicle con ol
• G oup 3: T ea ed wi h CCl4 in oli e oil (2 ml/kg body weigh ) (nega i e con ol)
• G oup 4: T ea ed wi h CCl4 in oli e oil (2 ml/kg) + ex ac (250 mg/kg)
• G oup 5: T ea ed wi h CCl4 in oli e oil (2ml/kg) + ex ac (500 mg/kg)
• G oup 6: T ea ed wi h CCl4 in oli e oil (2ml/kg) + ex ac (1000 mg/kg).
• G oup 7: T ea ed wi h CCl4 in oli e oil (2ml/kg) + i amin E (100 mg/kg) (S anda d d ug)
2.3. T ea men o samples
A he end o 21 days o ea men wi h he ui ex ac s, he animals we e sac i iced by ce ical capi a ion. Blood was
ob ained h ough ca diac punc u e by means o hypode mal sy inge and needle. The collec ed blood samples we e
placed in ice-cold mic o cen i uge ubes. The blood was allowed o coagula e and cen i uged a 4000 /min o 10 min.
The se um samples we e collec ed and u ilized o biochemical pa ame e s analysis.
2.3.1. Se um biochemical analysis:
Hepa ic biochemical es s (AST, ALT, and ALP) we e pe o med on a clinical chemis y au oma ic analyze (ADVIA 2400,
Baye Diagnos ics). AST, ALT, ALP, o al p o ein, albumin, we e measu ed acco ding o he p e ious me hods [22, 23]
using comme cially a ailable assay ki s (Baye Diagnos ics).
2.3.2. His opa hological s udy
His ological s udy o he li e specimen was ca ied ou using he s anda d me hod [24]. The li e was ins an ly
dissec ed om he animals a e sac i icing and insed wi h no mal saline. The li e issues we e ea ed indi idually
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and ixed in 10% o malin o his ological analysis. Using a mic oscope, a pa hologis who was unawa e o he s udy’s
p ocedu e inspec ed he mic oscopic slides [25].
2.3.3. S a is ical analysis
Da a ob ained om he s udy we e analyzed using S a is ical Package o Social Sciences (SPSS-25). Resul s we e
p esen ed as mean ± s anda d e o o mean (SEM) o sample eplica es. Raw da a we e subjec ed o one-way analysis
o a iance (ANOVA) ollowed by pos hoc u key’s es . P<0.05 was conside ed s a is ically signi ican .
3. Resul s
3.1. Yield o ex ac
The weigh o he V. doniana ui a e he emo al o he seeds was 765g and he c ude ex ac ob ained om i was
333g. The pe cen age yield was 43.8%.
3.2. Acu e oxici y es (LD50) o he ex ac :
O al adminis a ion o he ui ex ac up o 5000 mg/kgbw dose p oduced no changes in beha io , nei he was he e
any mo ali y in any o he g oups. The e o e, he LD50 o he ui ex ac o Ve ex doniana was abo e 5000 mg/kgbw.
3.3. Phy ochemical sc eening o he ex ac :
Phy ochemical analysis o he ex ac e ealed abundan p esence o alkaloids, annins, la onoids, glycosides, saponins,
s e oids, mode a e amoun o a s, acidic compounds and an h aquinone, and aces o educing suga s (Table 1).
Table 1 Pho ochemical sc eening o ex ac
Cons i uen s
Alk
Tan
Fla
Gly
Sap
An h
S e
Fa s
Acidic cpds
Red suga s
P esence
+++
+++
+++
+++
+++
++
+++
++
++
+
Key: Alk = alkaloids, Tan= annins, Fla = la onoids, Gly= glycosides, An h = an h aquinones, Sap=saponin, S e= s e oids, Red suga = educing
suga . (++) = mode a ely p esen , (+++) = abundan .
3.4. E ec s o ex ac s on li e unc ion pa ame e s
A he end o 21-days o ea men s wi h V. doniana ui ex ac , he e was a dose dependen signi ican (P<0.05)
educ ion in he le els o ALT, AST and ALP enzymes in he ea ed g oups when compa ed o he nega i e con ol g oup
(Table 2). The ui ex ac was also able o signi ican ly (P<0.05) inc ease he se um o al p o ein le el when compa ed
o he nega i e con ol g oup (Table 2).
Table 2 E ec s o he ex ac on li e unc ion pa ame e s
G oup (n=5)
ALT (U/I)
AST(U/I)
ALP(U/I)
To al p o ein (g/L)
Albumin (g/L)
No mal con ol
14.50 ± 2.5*
46.11 ±1.45*
85.37±0.45*
69.14±0.40 *
39.50±1.46 *
Vehicle con ol
14.78±1.43 *
45.55±2.30
82.88±1.37
66.15±0.27
39.99±0.21*
Nega i e con ol
39.42 ± 0.43
89.00±1.44
147.20±1.55
33.60±1.23
20.12±1.4
Posi i e con ol
14.36 ± 0.19*
46.29±0.12 *
87.59±0.22 *
68.27±0.88 *
38.33±0.67 *
250 mg/kg ex ac
20.12 ± 0.23*
55.55±1.29 *
95.22±1.53 *
59.11±1.59 *
25.67±0.18 *
500 mg/kg ex ac
16.21±1.11*
49.32±0.42 *
88.44±0.47 *
64.33±0.19 *
49.59±1.33 *
1000mg/kg ex ac
14.11±0.72 *
45.79±0.33 *
85.12±1.36 *
68.67±1.14 *
45.12±0.55 *
Values a e ep esen ed as S anda d e o o mean (SEM) (n=5). *p<0.05: S a is ically signi ican ly di e en om he con ol g oup
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3.5. His opa hological s udy o he li e
The li e s o he no mal con ol and ehicle con ol g oups (Figu es A and B, espec i ely) e ealed no mal ea u es o
he li e . Howe e , he e was ex ensi e degene a ion o hepa ocy es wi h ocal nec osis, acuola ed cy oplasm (shown
by he a ow), in lamma o y cell in il a ion and damaged cen al ein in he nega i e con ol g oup (Figu e 2C). On he
o he hand, he ui ex ac demons a ed a dose-dependen es o a ion o he li e a 100, 250, 500 and 1000 mg/kg
doses (Figu es 2D, 2E, 2F, and 2G espec i ely). A he end o he 21-day ea men , almos no mal appea ance o he
hepa ocy es a ound he cen al ein was obse ed in he 1000 mg /kg ea ed g oup (Figu e 2G).
Figu e 2 His opa hological s udy o he li e
4. Discussion
The li e is a i al o gan ha plays a c ucial ole in he main enance o homeos asis. I is in ol ed in de oxi ica ion,
me abolism, s o age and p oduc ion [26]. Ca bon e achlo ide (CCl4) is a well-known hepa o oxin capable o causing
li e damage [5]. The changes linked wi h CCl4-induced li e damage a e simila o ha o acu e i al hepa i is,
d ug/chemicals-induced hepa opa hy and oxida i e s ess. Ca bon e achlo ide-induced hepa o oxici y model is
commonly used o he e alua ion o hepa op o ec i e e icacy o d ugs and plan ex ac s [27]. P ese ing he ypical
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(02), 989-996
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physiological p ocesses o he hepa ic o gans ha ha e been dis up ed by hepa o oxins is a solid c i e ion o e alua ing
he e ec i eness o any hepa op o ec i e he apy.
The ques ion o sa e y and oxici y places signi ican es ic ions on he usage o he apeu ic he bs. This is because some
he bal medicines ha e been implica ed in o gan damage and a al e en s [28]. In his s udy he acu e oxici y s udy
(LD50) o Vi ex doniana ui ex ac e ealed absence o oxic signs and dea hs up o he dose o 5000 mg/kg in he a s.
Absence o oxic signs and dea hs in acu e oxici y s udy a e signs o ela i e sa e y.
The phy ochemical sc eening o he ui ex ac e ealed he abundan p esence o seconda y me aboli es such as
alkaloids, annins, la onoids, glycosides, saponins, s e oids and an h aquinone. Seconda y me aboli es ha e been
linked o he pha macological e ec s o plan ex ac (h ps://www. esea chga e.ne )
Vi ex doniana ui ex ac was able o signi ican ly (P < 0.05) es o e he ele a ed enzyme (AST, ALT and ALP) le els,
while inc easing he o al se um p o ein in a dose dependen manne when compa ed o he nega i e con ol. Due o
he des uc ion o hepa ic cell wall hese enzymes (AST, ALT and ALP) a e d ained in o he blood s eam. Inc ease in
hese enzymes sugges s hepa ic damage [29]. In con as , o al p o ein and albumin le els a e lowe ed in he p esence
o hepa ic damage [30]. The obse ed dose dependen signi ican (P< 0.05) dec ease in se um enzymes in he ea ed
g oups may be due o he abili y o he ui ex ac o p e en he leakage o in acellula enzymes h ough memb ane-
s abilizing ac i i y.
The esul o he his opa hology s udy o he li e s e ealed ex ensi e damage in he li e o he nega i e con ol g oup,
while he e was a dose-dependen es o a ion o he li e o a s ea ed wi h 100, 250, 500 and 1000 mg/kg o he o
he ex ac . A he end o he 21-day ea men , li e s o g oup ea ed wi h 1000 mg/kg o ex ac displayed almos
no mal appea ance o he hepa ocy es. This his ology esul he e o e u he suppo s he hepa op o ec i e e ec o
he ui ex ac o Vi ex doniana.
5. Conclusion
The ui ex ac o Vi ex doniana possesses hepa op o ec i e e ec agains ca bon e achlo ide induced li e damage,
acco ding o he indings o his s udy. Howe e , u he s udies a e equi ed wi h he aim o demons a ing he p ecise
molecula mechanisms o he hepa op o ec i e e ec s o Vi ex doniana and de e mining i s long- e m oxici y p o ile.
Compliance wi h e hical s anda ds
Disclosu e o con lic o in e es
The au ho s wish o con i m ha he e is no known con lic o in e es s associa ed wi h his pape and he e has been
no signi ican inancial suppo o his wo k ha could ha e in luenced i s ou come.
S a emen o e hical app o al
E hical app o al numbe PHACOOU/AREC/2023/032 was assigned o a es he animals we e ca ed o acco ding o
he Facul y o Pha macy (COOU) Animal Resea ch E hics Commi ee Guidelines (PHACOOUAREC), which a e in line wi h
he Na ional Ins i u e o Heal h (NIH), USA, guidelines o he ca e and use o labo a o y animals.
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