Co esponding au ho : U hman Ademola Yusu
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.
Amelio a i e e ec o bee oo aqueous ex ac on he li e o diabe ic Wis a a s
Ma y kabole 1, Isabel Nam ukwe Luambia 1, Albe ina Ng’andu 2, Olawole Temi ayo P iscilla 3 and U hman
Ademola Yusu 1, *
1 Depa men o Human Ana omy, School o Medicine and Heal h Sciences, Mulungushi Uni e si y, Li ings one Campus,
Zambia.
2 Depa men o Ana omy, Ins i u e o Basic and Biomedical Sciences, Le y Mwanawasa Medical Uni e si y, Zambia.
3 Depa men o Ana omy, College o Medicine, Uni e si y o Lagos, Nige ia.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 26(02), 2335-2343
Publica ion his o y: Recei ed on 04 Ap il 2025; e ised on 11 May 2025; accep ed on 13 May 2025
A icle DOI: h ps://doi.o g/10.30574/wja .2025.26.2.1860
Abs ac
This s udy in es iga ed he e ec s o bee oo ex ac on he li e o diabe ic Wis a a s. Thi y male Wis a a s (8-10
weeks old, weighing 160-200 g) we e andomly assigned o i e g oups: A) no mal con ol, B) diabe ic only, C) diabe ic
+ bee oo , D) diabe ic + Me o min, and E) bee oo ex ac only. Diabe es was induced ia a single in ape i oneal dose
o s ep ozo ocin (70 mg/kg body weigh ). A e 72 hou s o sus ained diabe es (blood glucose ≥ 7 mmol), he a s
ecei ed o al adminis a ions o bee oo ex ac and Me o min (500 mg/kg and 100 mg/kg body weigh daily
espec i ely) o ou weeks, while being ed a s anda d die wi h wa e access. Blood glucose le els and body weigh
we e measu ed weekly. A he expe imen 's conclusion, he a s we e eu hanised, and ela i e li e weigh s we e
eco ded. Da a analysis included one-way ANOVA and g aphical ep esen a ion using Excel. Resul s indica ed
signi ican di e ences in blood glucose le els and a e age body weigh in he diabe ic a s (p<0.05) compa ed o o he
g oups. The ela i e li e weigh in he diabe ic g oup was no ably lowe (p<0.05). His ological examina ion e ealed
no mal li e a chi ec u e in he con ol and bee oo g oups, whe eas he diabe ic g oup exhibi ed dis up ed
his oa chi ec u e wi h nec o ic hepa ocy es. The diabe ic + bee oo g oup showed no malised his oa chi ec u e
simila o he con ol g oup, while he diabe ic + Me o min g oup displayed mino dis up ions. All g oups excep he
diabe ic g oup displayed no mal eac ions o PAS s aining. In conclusion, bee oo ex ac demons a ed
hepa op o ec i e e ec s, mi iga ing diabe es- ela ed li e damage and weigh loss in male Wis a a s.
Keywo ds: S ep ozo ocin; Li e ; Diabe es; Wis a Ra s; Bee oo
1. In oduc ion
Diabe es melli us is a medical condi ion cha ac e ised by abno mal me abolism o p o eins, ca bohyd a es, and a s,
leading o high blood glucose and lipid le els. The mos common sub ypes a e ype 1 and ype 2 diabe es melli us,
hough o he o ms exis [1]. Type 1 diabe es a ec s 5-10% o indi iduals wi h diabe es and is caused by dys unc ion o
be a cells, esul ing in dec eased insulin p oduc ion and low le els o ci cula ing insulin [2]. Symp oms ypically
mani es du ing childhood o adolescence bu can appea la e in li e as well. Con e sely, ype 2 diabe es melli us
accoun s o abou 90% o 95% o diabe es cases globally [2]. I is p ima ily ma ked by insulin esis ance in pe iphe al
issues and an insu icien insulin esponse. Obesi y, especially abdominal a accumula ion, is a common ai among
pa ien s wi h ype 2 diabe es. Bo h gene ic and en i onmen al ac o s play a ole in i s epidemiology, wi h gene ics
becoming inc easingly signi ican in he con ex o seden a y li es yles and high-calo ie die a y habi s [3]. Key o gans
in ol ed in de eloping ype 2 diabe es include he li e , skele al muscle, kidneys, b ain, small in es ine, adipose issue,
and panc eas. Less common o ms o diabe es include ges a ional diabe es, ma u i y-onse diabe es o he young, la en
au oimmune diabe es in adul s, and seconda y diabe es, which can esul om o he medical condi ions, such as
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panc ea i is, o as a side e ec o medica ions like co icos e oids [4]. The li e , comp ising abou 2.5% o he body
weigh , is i al o nu ien me abolism and was e elimina ion [5]. I plays an essen ial ole in p oducing bile o lipid
diges ion and abso p ion in he small in es ine and egula ing coagula ion h ough he manu ac u e o i amin K and
clo ing ac o s [6]. Recen ly, he e has been inc easing in e es in he bal and complemen a y he apies, pa icula ly
hose u ilising medicinal plan s ich in an ioxidan s, o help manage diabe es by educing in lamma ion [7]. Bee oo
has eme ged as a popula unc ional ood due o i s a ious biological ac i i ies and po en ial he apeu ic bene i s
agains condi ions such as hype ension, dep ession, in lamma ion, in ec ions, and diabe es [8]. Some s udies indica e
ha bee oo ex ac may possess hepa op o ec i e and an i-diabe ic p ope ies [9]. Consequen ly, his s udy aims o
e alua e he e ec s o bee oo ex ac on he li e o Wis a a s wi h induced diabe es melli us.
2. Ma e ial and me hods
2.1. Plan ma e ials
The Bee oo was ha es ed om he Li ings one dis ic , Sou he n P o ince o Zambia. I was subjec ed o
iden i ica ion a he Uni e si y o Zambia, School o Na u al Sciences, unde he Depa men o Biological Sciences be o e
he s udy began. The Bee oo was ai -d ied and pounded. The d y pounded Bee oo was hen g ound and sie ed o
ob ain a homogeneous powde . The ex ac ion was done using me hods desc ibed by Bo jan e al [10].
2.2. Animals and animal managemen
Thi y adul , p esumably heal hy male Wis a a s (Ra us no egicus) we e used o his s udy. The animals we e
be ween 8 o 10 weeks old, wi h a body weigh o 160-200 g. They we e kep in i e cages (6 a s pe cage) and housed
in he animal holdings o he Depa men o Ana omy, Mulungushi Uni e si y School o Medicine and Heal h Sciences.
The a s we e main ained on s anda d animal eeds (Weal h-ga e pelle ized eeds) and allowed ee access o clean
wa e and eed (ad libi um).
2.3. Induc ion o diabe es.
S ep ozo ocin (STZ) was used o induce diabe es. Ra s we e weighed, and a baseline glucose le el was es ablished
ollowing an o e nigh as ing pe iod. The animals we e injec ed wi h s ep ozo ocin, calcula ed a a dose o 70 mg/kg
body weigh , and hen e u ned o hei no mal eeding cycle [11]. I ook abou 72 hou s o diabe es o be es ablished
in he animals ollowing he adminis a ion o s ep ozo ocin. Subsequen ly, a as ing blood sample was collec ed o
con i m he es ablishmen o diabe es using a ail ein punc u e. A glucome e was used o assess blood glucose le els,
wi h animals conside ed diabe ic i as ing blood glucose le els exceeded 7 mmol/L (≥ 250 mg/dL).
2.4. Expe imen al design
Thi y adul heal hy male Wis a a s we e di ided in o i e g oups o six (6) Wis a a s each. Con ol G oup A consis ed
o no moglycemic animals ha ecei ed nei he STZ no bee oo ex ac . G oup B was diabe ic wi hou ecei ing
bee oo ex ac o me o min. G oup C was diabe ic ea ed wi h bee oo ex ac only. G oup D was diabe ic ea ed
wi h Me o min only, and G oup E ecei ed bee oo ex ac only.
2.5. Bee oo mode o adminis a ion
The dose o he aqueous ex ac s o Bee oo used in hese s udies was adop ed om a epo by [12]. Bee oo was
dissol ed in physiological saline daily and adminis e ed o ally using an o o-gas ic cannula o G oup C a s (n=6) a a
dosage o 500 mg/kg body weigh (bw) a 9:00 – 10:00 a.m. each day o a maximum pe iod o ou weeks. G oup D
(n=6) ecei ed 100 mg/kg bw o me o min, and G oup E a s (n=6) we e adminis e ed 500 mg/kg bw o bee oo
ex ac s. G oup A a s (n=6) ecei ed nei he STZ no bee oo aqueous ex ac .
2.6. Measu emen o blood glucose
Blood glucose le els we e e alua ed a e o e nigh as ing he a s a 9:00 – 10:00 hou s, using he Glucose oxidase
me hod wi h a One Touch Ul a 2 glucome e s (Accu-Chek Compac Plus). Blood samples we e ob ained om he
median caudal ein o he ail by snipping he ip. The blood glucose le el was moni o ed weekly o one week
(acclima isa ion pe iod) be o e he induc ion o diabe es and moni o ed o ou weeks du ing ea men [13].
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2.7. Measu emen o body weigh (g)
Body weigh (g) o he a s was eco ded o one week (acclima isa ion pe iod) be o e he induc ion o diabe es and
e e y week du ing he expe imen al ea men o ou weeks. Body weigh was measu ed using a weighing scale
(Venus VT 30 SL) [13].
2.8. The ela i e o gan weigh (%)
The ela i e weigh o he li e o he a s was e alua ed as he a io o he li e 's espec i e weigh o he e minal body
weigh o he same a , eco ded as a pe cen age (%) using a sensi i e weighing balance (SonyF3G b and) [13].
2.9. His ological p ocess.
A he end o he s udy, animals we e sac i iced by eu hanasia. They we e laid supine on he dissec ing boa d and pinned
h ough he o e and hind paws. The abdomen o he animals we e dissec ed using scalpel and su gical blade, and each
o gan was ca e ully emo ed and weighed. Tissues we e ixed in eshly p epa ed o mo saline o 72 hou s and
p ocessed o ou ine his ological examina ions, s ained wi h Haema oxylin and Eosin (H&E) o obse e changes in
cellula mo phology. Pe iodic acid-Schi (PAS) s ain was also used o checked o he p esence o he glycogen.
2.10. Pho omic og aphy
Pho omic og aphs o his ological sec ions o he li e we e aken wi h an Olympus Mic oscope (New Yo k, Uni ed S a es
o Ame ica) coupled wi h a came a a he Depa men o Human Ana omy, Mulungushi Uni e si y School o Medicine
and Heal h Sciences, Li ings one Campus, Zambia.
2.11. Da a analysis.
Da a we e p esen ed as mean ± s anda d e o o he mean (mean ± SEM) and analysed using one-way ANOVA. All
g aphs we e d awn using Excel (Mic oso Co po a ion, U.S.A.). P alues less han 0.05 (p < 0.05) we e conside ed
s a is ically signi ican .
3. Resul s
3.1. A e age blood glucose le els on weekly basis
Figu e 1 Blood glucose le els o Wis a a s on weekly basis. Da a we e exp essed as mean ± SEM (p<0.05)
The a e age blood glucose le els we e moni o ed weekly ac oss di e en g oups o Wis a a s. Du ing he
acclima isa ion week (week-1), all a s we e no moglycemic. The con ol and bee oo -only g oups main ained s able
blood glucose le els h oughou he s udy. In week 0, which was he induc ion week, he diabe ic g oup, as well as he
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diabe ic + bee oo and diabe ic + me o min g oups, showed ele a ed blood glucose le els. By week 1, he diabe ic +
bee oo g oup began o show a dec ease in blood glucose le els, which was s a is ically signi ican (p<0.05) when
compa ed o he con ol and bee oo -only g oups. A simila dec ease was obse ed in he diabe ic + me o min g oup.
In week 3, he diabe ic + bee oo g oup's blood glucose le els we e app oaching he no mal ange, and when compa ed
o he con ol and bee oo -only g oups, he di e ences we e no longe s a is ically signi ican (p>0.05). In con as , he
diabe ic + me o min g oup s ill exhibi ed s a is ically signi ican di e ences (p<0.05). By week 4, all g oups e u ned
o no mal blood glucose le els excep he diabe ic g oup.
3.2. A e age body weigh on weekly basis
A he s a o he expe imen , he acclima isa ion week (week-1), he a e age body weigh o all he Wis a a s om
he a ious g oups was nea ly iden ical, wi h only a negligible di e ence obse ed. In week 0, du ing he induc ion
phase, he con ol g oup showed he highes inc ease in weigh , ollowed closely by he me o min g oups, hough no
signi ican di e ences we e no ed ac oss all g oups. By week 2, a g adual dec ease in weigh was obse ed in he
diabe ic g oup, while he o he g oups con inued o show weigh inc eases. The compa ison be ween he diabe ic and
con ol g oups was s a is ically signi ican (p<0.05), bu he compa ison wi h he bee oo and me o min g oups did
no show signi ican di e ences (p>0.05). In week 3, he weigh dec ease in he diabe ic g oup became s a is ically
signi ican (p<0.05) compa ed o all o he g oups, and his end con inued up o week 4, indica ing a consis en
ela ionship h oughou he expe imen .
Figu e 2 A e age body weigh on weekly basis. Da a we e exp essed as mean ± SEM (p<0.05)
3.3. Rela i e weigh o he li e (%)
Figu e 3 p esen s he ela i e weigh o he li e among di e en g oups. The indings indica e ha he diabe ic g oup
had he leas li e weigh , ollowed by he diabe ic + me o min g oup, he diabe ic + bee oo g oup, he bee oo -only
g oup, and inally he con ol g oup. The e was a s a is ically signi ican di e ence in he ela i e li e weigh o he
diabe ic g oup compa ed o all o he g oups (p<0.05). Howe e , he di e ences be ween he diabe ic + bee oo and
diabe ic + me o min g oups compa ed o he con ol and bee oo -only g oups we e no s a is ically signi ican
(p<0.05).
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Figu e 3 blood glucose le els o Wis a a s on weekly basis. Da a we e exp essed as mean ± SEM (p<0.05)
3.4. His ology o he li e
The li e in he no mal con ol and bee oo g oups exhibi ed no mal his oa chi ec u e wi h nume ous heal hy
hepa ocy es (Fig. 4 A and E). In con as , he diabe ic g oup displayed dis up ed his oa chi ec u e cha ac e ised by
many nec o ic hepa ocy es (Fig. 4 B). The diabe ic+bee oo g oup demons a ed a his oa chi ec u e simila o ha o
he con ol g oup (Fig. 4 C), while he diabe ic+me o min g oups exhibi ed minimal dis up ion, con aining bo h no mal
and nec o ic hepa ocy es (Fig.4 D). The no mal con ol, bee oo only, diabe ic+bee oo , and diabe ic+me o min
g oups showed a no mal eac ion (Fig. 5 A, C, D, and E), while he diabe ic g oup displayed a posi i e eac ion o he PAS
s ain (Fig.5 B).
Figu e 4 Pho omic og aph showing he Li e a day 28. H&E s ain X400. A- No mal con ol, B – Diabe ic, C –
Diabe ic+bee oo , D – Diabe ic+Me o min and E- Bee oo only. A ow – hepa ocy e, A ow head – Nec o ic
hepa ocy e
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Figu e 5 Pho omic og aph showing he Li e a day 28. PAS s ain X400. A- No mal con ol, B – Diabe ic, C –
Diabe ic+Bee oo , D – Diabe ic+Me o min and E- Bee oo only. A ow – hepa ocy e, deep colo a ion indica es
posi i e PAS eac ion
4. Discussion
Diabe es melli us has been associa ed wi h mul iple issue damage and signi ican ly con ibu es o he de elopmen o
non-alcoholic a y li e disease (NAFLD). In con as , bee oo is gaining a en ion o i s hepa op o ec i e and
hypolipidemic p ope ies in a s and abbi s subjec ed o high-choles e ol die s [12]. This s udy aimed o e alua e he
e ec s o bee oo aqueous ex ac on he li e o Wis a a s wi h diabe es induced by s ep ozo ocin.
As shown in Figu e 1, Wis a a s in he diabe ic g oup exhibi ed pe sis en ly ele a ed blood glucose le els h oughou
he expe imen . This ele a ion is a ibu ed o he damage in lic ed by s ep ozo ocin on panc ea ic be a cells, esul ing
in inadequa e insulin p oduc ion [9]. Rema kably, in he diabe ic + bee oo g oup, blood glucose le els began o
no malise by week 3 and achie ed no moglycemia by week 4. This imp o emen can be linked o he insulin
sensi isa ion, an ioxidan ac i i y, and hypoglycemic e ec s o bee oo [12]. Con e sely, he me o min- ea ed g oup
emained diabe ic un il week 3 and only eached no moglycemia in week 4, indica ing ha bee oo 's e icacy may s em
om i s highe concen a ion o bene icial biochemical cons i uen s [14]. Nume ous s udies ha e unde sco ed
bee oo 's abili y o enhance ni ic oxide p oduc ion, a compound ha is o en de icien in insulin- esis an diabe es
[15]. Re aa and El-Nassag [16] also highligh ed ha ni ic oxide imp o es insulin signalling and p omo es glucose
anspo e 4, sugges ing ha inc easing ni ic oxide could e ec i ely imp o e insulin sensi i i y and educe blood
glucose le els. Fu he mo e, Yusu e al. [17] epo ed bee oo 's capaci y o s imula e insulin p oduc ion and sec e ion
h ough be a cell ac i a ion by be anin, aligning wi h he indings o he p esen s udy.
In Figu e 2, he diabe ic g oup showed a p og essi e weigh loss om week 3 un il he end o he s udy. This
phenomenon is due o diabe es impai ing glucose u ilisa ion by cells, p omp ing a ca abolic esponse in ol ing a and
p o ein deg ada ion, ul ima ely leading o muscle was ing [9]. Con e sely, he diabe ic + bee oo g oup main ained
hei body weigh h oughou he expe imen , likely due o bee oo 's abili y o enhance insulin sensi i i y in cells, hus
p e en ing eliance on adipose issue and muscle p o ein o ene gy [18]. Al hough he diabe ic + me o min g oup
displayed a diminished a e o weigh gain a e week 3, hei o e all weigh inc eased, highligh ing ha me o min has
a limi ed impac on pe iphe al insulin sensi i i y [19]. These indings co obo a e he obse a ions o Pang e al. [20],
who no ed ha inadequa e insulin sec e ion om panc ea ic be a cells educed glucose up ake in cells, causing ene gy
sou cing om issue p o eins and, consequen ly, weigh loss. Addi ionally, as epo ed by Pu e al. [21], me o min's
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ac ion does no inhibi body weigh educ ion and is being explo ed as a po en ial weigh loss agen . Bee oo 's con en
o be alains, which p e en glycogenolysis and imp o e blood glucose le els, also plays a ole in inhibi ing adipose issue
b eakdown [17]. Figu e 3 illus a es ha he diabe ic g oup had he lowes ela i e li e weigh , indica ing li e damage
caused by diabe es [4]. Howe e , he diabe ic + bee oo g oup did no show a s a is ically signi ican di e ence in li e
weigh compa ed o he con ol g oup (p<0.05). This may be a ibu ed o bee oo 's p o ec i e e ec s agains oxida i e
s ess and i s ole in educing lipid accumula ion, he eby p ese ing li e in eg i y [12]. These indings con as wi h
hose epo ed by Eluehike and Onoagbe [22], who ound an inc ease in li e weigh among hei diabe ic subjec s.
Howe e , ou esul s align wi h Abd El-Gh a e al. [7], who obse ed ha bee oo posi i ely in luenced anabolic
p ocesses and imp o ed bo h glucose and lipid me abolism, con ibu ing o i s hepa op o ec i e p ope ies.
His ological assessmen o he li e u ilising H&E s aining e ealed no mal his oa chi ec u e in bo h he con ol and
bee oo g oups, cha ac e ised by abundan hepa ocy es. The diabe ic + bee oo g oup also exhibi ed a simila
his ological p o ile, sugges ing ha bee oo e ec i ely econs uc ed he li e and p e en ed u he damage [23]. In
con as , he diabe ic + me o min g oup displayed mino dis up ions in hei li e a chi ec u e, wi h bo h no mal and
nec o ic hepa ocy es. This disc epancy indica es ha me o min may no adequa ely epai li e damage esul ing om
diabe es melli us [19]. The diabe ic g oup demons a ed signi ican a chi ec u al dis up ion wi h nume ous nec o ic
hepa ocy es, a ibu able o he oxic e ec s o diabe es on he li e [18]. These esul s a e consis en wi h p io s udies
documen ing he hepa op o ec i e e ec s o bee oo , which has been shown o educe li e enzyme le els and lipid
accumula ion. Ele a ed blood glucose le els lead o he gene a ion o eac i e oxygen species (ROS), causing oxida i e
s ess ha igge s in lamma ion and, ul ima ely, hepa ocy e damage [24]. Simila ly, Mahmoud Abbas [9] con i med
ha bee oo p o ec s he li e by i s high concen a ions o polyphenols and la onoids, which impa po en
an i adical p ope ies. Findings pa allel o hose om Sui e al. [25] e ealed ha me o min has minimal e ec s on lipid
le els and hepa ocy e degene a ion, consis en wi h ou obse a ions. In he PAS eac ion assessmen , he con ol,
bee oo -only, diabe ic + bee oo , and diabe ic + me o min g oups exhibi ed no mal esponses, while he diabe ic
g oup demons a ed a posi i e PAS eac ion. This esponse indica es inc eased glycogen accumula ion due o diabe es
melli us [18], aligning wi h indings by Ashc o e al. [26] and [27] which a ibu ed ele a ed glycogen le els o
impai ed glycogenolysis, glycosyla ion o p o eins, and issue damage esul ing om hype glycemia.
5. Conclusion
This s udy es ablished he hepa op o ec i e e ec s o bee oo , e idenced by educ ions in blood glucose le els and he
p ese a ion o no mal his oa chi ec u e in li e issue. Addi ionally, bee oo played a signi ican ole in p e en ing
body weigh loss in diabe ic Wis a a s. These indings sugges ha bee oo has he po en ial o mi iga e diabe es-
ela ed mo ali y and mo bidi y.
Compliance wi h e hical s anda ds
Acknowledgmen s
I app ecia e M Alick Tembo and Josipha Chizambe o helping us o eed he animals.
Disclosu e o con lic o in e es
No con lic o in e es o be disclosed.
S a emen o e hical app o al
The e hical app o al and pe mission o he s udy was ob ained om Mulungushi Uni e si y School o Medicine and
Heal h Sciences Resea ch e hic commi ee
Re e ences
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