Zoology
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Item Kaempferol alleviates neurodegenerative disorders induced by Naja nigricollis venom via mechanisms of antioxidants, anti-inflammatory, dopaminergic and neuronal functions(Elsevier, 2024) Ajisebiola, B.S.; Mustapha, A-R.K.; Oyedara, O.O.; Oladele, J.O.; Adeyi, A.O."Naja nigricollis venom (NnV) contains neurotoxins that influence neurological functions. Kaempferol is a bioactive compound present in edible plants with numerous pharmacological activities. This study investigated the ameliorative potential of kaempferol against NnV-induced neurotoxicity in rats. Fifty male Wistar rats were randomized into five groups (n =10). Group 1 rats were the control while 1.0 mg/kg1 (LD50) of NnV was injected intraperitoneally into rats in groups 2–5 to observed neurotoxicity. Group 2 was untreated post en- venomation, while groups 3–5 were treated with polyvalent antivenom, 4 and 8 mg/kg of kaempferol, respec- tively. The biochemical analysis, neurotoxicity, and pathomorphological defects were assessed in the brain of the envenomed treated rats. Envenomation with NnV elevated oxidative and inflammatory biomarkers, and induced neurotoxicity accompanied with neurobehavioral deficits, and severe pathohistological defects were seen in the brain of untreated envenomed rats. However, treatment with kaempferol significantly (p <0.05) decreased malondialdehyde (MDA) levels and upregulated levels of reduce glutathione (GSH) antioxidant including su- peroxide dismutase (SOD) and glutathione peroxidase (GPX) antioxidant enzymes, while inflammatory bio- markers; nitric oxide (NO) levels and myeloperoxidase (MPO) activity significantly decreased in envenomed treated groups. Kaempferol upregulated dopamine concentration with significant suppression of acetylcholin- esterase (AchE) activity, and restored neurobehavioral and locomotor activities in envenomed treated rats. Also, severe pathomorphological alterations observed in the cortex of the brain were attenuated after kaempferol treatment. The underlaying ameliorative mechanisms of kaempferol are linked to its antioxidant activity, lipid peroxidation inhibition, anti-inflammatory activity, acetylcholinesterase suppression, and alleviation of dopa- mine system and neurobehavioral abilities."Item Kaempferol mitigates reproductive dysfunctions induced by Naja nigricollis venom through antioxidant system and anti‑inflammatory response in male rats(Nature Portfolio / Springer Nature, 2024) Adeyi, A.O.; Ajisebiola, B.S.; Sanni, A.A.; Oladele, J.O.; Mustapha, A-R.K.; Oyedara, O.O.; Fagbenro, O.S.Naja nigricollis Venom (NnV) contains complex toxins that affects various vital systems functions after envenoming. The venom toxins have been reported to induce male reproductive disorders in envenomed rats. This present study explored the ameliorative potential of kaempferol on NnVinduced male reproductive toxicity. Fifty male wistar rats were sorted randomly into five groups (n = 10) for this study. Group 1 were noted as the control, while rats in groups 2 to 5 were injected with LD50 of NnV (1.0 mg/kg bw; i.p.). Group 2 was left untreated post envenomation while group 3 was treated with 0.2 ml of polyvalent antivenom. Groups 4 and 5 were treated with 4 and 8 mg/ kg of kaempferol, respectively. NnV caused substantial reduction in concentrations of follicle stimulating hormone, testosterone and luteinizing hormone, while sperm motility, volume and counts significantly (p < 0.05) decreased in envenomed untreated rats. The venom enhanced malondialdehyde levels and substantially decreased glutathione levels, superoxide dismutase and glutathione peroxidase activities in the testes and epididymis of envenomed untreated rats. Additionally, epididymal and testicular myeloperoxidase activity and nitric oxide levels were elevated which substantiated severe morphological defects noticed in the reproductive organs. However, treatment of envenomed rats with kaempferol normalized the reproductive hormones with significant improvement on sperm functional parameters. Elevated inflammatory and oxidative stress biomarkers in testis and epididymis were suppressed post kaempferol treatment. Severe histopathological lesions in the epididymal and testicular tissues were ameliorated in the envenomed treated groups. Results highlights the significance of kaempferol in mitigating reproductive toxicity induced after snakebite envenoming.Item Biosynthesis, characterization and antivenom activities of Moringa oleifera silver nanoparticles: an experimental approach(Zoological Society of Nigeria, 2023) Adeyi, A.O.; Olowookorun, T.O.; Ajisebiola, B.S.; Labulo, H.A.; Adeyi, O.E.; Ibrahim, H.Moringa oleifera has been previously established to possess neutralizing potentials against Echis ocellatus venom. This study however, investigated the bioefficacy of silver nanoparticles biosynthesized from M. oleifera leaf extract aimed at improving its bioactivity against E. ocellatus venom-induced toxicities using in vivo and in vitro methods. The intrinsic characteristics of the produced M. oleifera-Silver nanoparticles (MO-AgNPs) were carried out using energy dispersive X-ray, X-ray diffraction, scanning electron microscopy, transmission electron microscopy and Fourier-transform infrared spectroscopy. Twenty-five male Wistar rats divided randomly into five groups (n=5) were used for the antivenom study. Group 1 received saline while groups 2 to 5 were envenomed intraperitoneally with 0.22mg/kg (LD50) of E. ocellatus venom. Group 2 was left untreated while groups 3 to 5 were treated with 0.2ml of antivenom, 5 and 10mg/kg MO-AgNPs post-envenomation, respectively. Blood and tissue of treated rats were analyzed for heamatological parameters and histopathology, respectively. The MO-AgNPs formation was confirmed with a colour change from light brown to yellowish-brown with maximum SPR band at 420nm from UV-Vis analysis, indicating a reflection of the bio-reduction of Ag+ to Ag0. The Transmission electron micrographs showed well dispersed spherical AgNPs with average particle size of 15.7nm. Treatment with MO-AgNPs caused a significant improvement of acute anemia, leucopenia and thrombocytopenia induced by the venom in the envenomed treated rats. Also, MO-AgNPs inhibited the haemorrhagic, haemolytic and anticoagulant activities of the venom. Tissue lesions observed in heart of envenomed untreated rats were attenuated after treatment with MO-AgNPs. The biosynthesized MO-AgNPs exhibited potent neutralizing potentials than M. oleifera crude extract against E. ocellatus venom toxicities.Item "Inhibition of Metalloproteinase from Bitis arietans Venom In Vitro by the Ethyl Acetate Fraction of Moringa oleifera and Its Chemical Profile"(Springer Nature, 2023) Ajisebiola, B.S.; Bello, F.T.; Osamudiamen, P.M.; Oladele, J.O.; Adeyi, A.O.Moringa oleifera Lam., Moringaceae, ethyl acetate soluble fraction has been documented to contain bioactive antivenom chemicals against Bitis arietans venom toxicities. Snake venom metalloproteinase is the principal enzyme of B. arietans venom exhibiting severe haemorrhage and haemostatic disturbances in envenomed victims. In this current study, B. arietans snake venom metalloproteinase was isolated using ion-exchange chromatography and gel filtration while SDS-PAGE was used to determine enzyme molecular weight. M. oleifera ethyl acetate soluble fraction was fractionated using column and thin-layer chromatography into subfractions and tested against B. arietans snake venom metalloproteinase. EchiTab polyvalent antivenom was used as the standard drug. The compounds present in the most active subfraction were identified using GCMS. The molecular weight of isolated enzyme was 43.28 kDa with specific activity of 3.06 μmol/min/mg and percentage yield of 18.5%. A total of 8 subfractions were obtained after fractionation and labelled ethyl acetate subfractions F1-8. All subfractions showed significant (p < 0.05) inhibition against B. arietans snake venom metalloproteinase activity. However, subfraction F5 demonstrated the highest inhibition against B. arietans snake venom metalloproteinase activity with total inhibition at all concentrations compared to other subfractions and antivenom. Subfraction F5 displayed substantial inhibition against the haemorrhagic, haemolytic, and anticoagulant activities of B. arietans venom with a dose-dependent effect. The major bioactive compounds of subfraction F5 were γ-sitosterol (5.92%), kaempferol (5.9%), quercetin (5.28%), n-hexadecanoic acid (5.23%), and n-pentacosane (5.22%) of which some were previously reported as potential snake venom inhibitors. Results showed that subfraction F5 possesses potential antivenom compounds that could be explored for snakebite treatment.Item "Inhibition of phospholipase A2 from Naja haje and Naja nigricollis venoms by active fraction of Moringa oleifera leaves: in vitro and in silico methods"(Taylor & Francis, 2023) Adeyi, A.O.; Jimoh, A.O.; Ajisebiola, B.S.; Adeyi, O.E.; Metibemu, D.S.; Okonji, P.E.Phospholipases are one of the principal toxic enzymes in snake venoms inducing a wide variety of pharmacological effects after envenomation. Natural inhibitors from plants are known to inhibit the toxic enzyme activities of snake venoms. In this study, ethanol crude extract of M. oleifera leaves was partitioned using n-hexane and ethyl acetate after which fractionation was done using column and thin layer chromatography. Subsequently, the inhibitory activities of the crude extract and sub-fractions of M. oleifera were investigated against phospholipases A2 isolated from Naja haje and Naja nigricollis venoms using in vitro and in-silico approaches while EchiTab-PLUS polyvalent antivenom was used as the standard drug. The molecular weight of isolated N. haje phospholipase A2 (NH-PL) and N. nigricollis phospholipase A2 (NN-PL) were 24.11 and 35.22 kDa respectively. NH-PL enzyme had a specific activity of 2.70 lM/min/mg substrate while NN-PL activity was 2.10 lM/min/mg substrate. The Km of NH-PL was 0.330 lM with Vmax of 0.085 lM/mL min while NN-PL had Vmax of 0.198 lM/mL.min and Km of 0.670 lM. M. oleifera nhexane sub-fraction 5 (MOLH5) exhibited a total inhibition of NN-PL and NH-PL enzyme activities at all concentrations used. Molecular docking of the phytoconstituents of MOLH5 against the catalytic site of phospholipase A2 revealed 2-Hydrazino-8-hydroxy-4-phenylquinoline as the lead compound and a potential drug candidate with a docking score of _6.789 kcal/mol. Findings indicated that MOLH5 possesses phospholipase A2 natural inhibitors that could be explored as a therapy for snake envenoming.Item "Ethyl Acetate Fraction of Moringa oleifera Leaves Mollified Toxicological Activities Actuated by Bitis arietans (Puff adder) Venom"("Faculty of Physical Sciences and Faculty of Life Sciences, Univ. of Ilorin, Nigeria", 2022) Ajisebiola, B.S.; Oyinlola, R.R.; Akintunde, S.O.; Paul, O.; Adeyi, A.O.Bitis arietans (Puff adder) venom possess numerous biological active toxins exhibiting toxicological actions after envenoming. Moringa oleifera crude extract has been documented to extenuate toxicities induced by B. arietans venom. This study reported the efficacy of n-hexane, ethyl acetate and ethanol solvent fractions obtained from crude extract of M. oleifera against the lethal dose toxicity, biological activities and oxidative stress induced by B.arietans venom to ascertain the best fraction with active antivenom phytochemicals. Forty-five male rats were randomly selected into nine groups (n=5) for the anti-oxidative stress study. Groups 1 served as control; group 2 to 9 were envenomed by a single intraperitoneal injection of 1.5 mg/kg (LD50) of the venom. Group 2 was not treated post envenomation while groups 3 was treated with polyvalent antivenom. Group 4, 6 and 8 were treated with 300 mg/kg while group 5, 7 and 9 were treated with 600 mg/kg of n-hexane, ethyl acetate and ethanol solvent fractions of M. oleifera respectively. High dose of M. oleifera ethyl acetate solvent fraction best neutralized the lethal dose toxicity and strongly inhibited the heamorrhagic and anticoagulant activities of B. arietans venom. The venom induced oxidative stress with significant (P<0.05) enhancement of Superoxide Dismutase, Catalase and Malondialdehyde levels in serum and heart tissues of untreated envenomed rats. However, ethyl acetate solvent fraction was most effective in normalizing the antioxidant enzyme activities of envenomed treated rats. Results showed that ethyl acetate fraction of M. oleifera possesses active antivenom phytochemicals against B. arietans venom induced toxicities.Item "Echis ocellatus Venom-Induced Reproductive Pathologies in Rat Model; Roles of Oxidative Stress and Pro-Inflammatory Cytokines"(MDPI, 2022) Ajisebiola, B.S.; Alamu, P.I.; James, A.S.; Adeyi, A.O.This study reported reproductive pathologies associated with Echis ocellatus venom in animal model. Twenty male Wistar rats with body weight between 180 and 220 g were selected randomly into two groups (n = 10). Rats in group 1 served as the control while rats in group 2 were envenomed with a single intraperitoneal injection of 0.055 mg/kg1 (LD6.25) of E. ocellatus venom on the first day and a repeated dose on the twenty fifth day. Both control and envenomed rats were monitored for fifty consecutive days. The venom caused a significant (p < 0.05) reduction in sperm motility, count, and volume, with increased sperm anomalies in envenomed rats compared to the control. Likewise, serum concentrations of male reproductive hormones were significantly (p < 0.05) higher in envenomed rats. Increased levels of malondialdehyde were accompanied by a significant (p < 0.05) decrease in reduced glutathione and catalase activity in the epididymis and testis tissues of envenomed rats. The venom enhanced the release of epididymal and testicular tumor necrosis factor-alpha and interleukin1-beta compared to the control. Furthermore, severe pathological defects were noticed in tissues of the testis and epididymis of envenomed rats. This study demonstrated that E. ocellatus venom toxins can induce reproductive dysfunction in male victims of snake envenoming.Item "An in vivo assessment of inflammatory and oxidative stress responses in Echis ocellatus-venom induced cardiotoxicity"(Elsevier, 2022) Ajisebiola, B.S.; Fawole, A.B.; Adeyi, O.E.; Adeyi, A.O.Echis ocellatus is one of the many viper species that accounts for severe pathophysiological alterations in tissues of organs after envenoming. However, limited information regarding the potential cardiac toxicity due to viper envenoming is available. This current study investigated cardiotoxicity associated with E. ocellatus envenoming in rat model. Twenty (20) male Wistar rats weighing between 140 and 180 g were divided randomly into two groups (n = 10). Rats in group 1 (control) were injected with saline while rats in group 2 were envenomed intraperitoneally with 0.055 mg/kg−1 (LD6.25) of E. ocellatus venom. The rats were envenomed on day 1 with a repeated dose administered on day 15, afterwards the animals were monitored till day 30. The venom caused significant (P < 0.05) reduction in body and heart weights including the heart index of envenomed rats compared to the control. Levels of malondialdehyde significantly (P<0.05) increased with decrease in glutathione concentration and catalase activity in heart tissues of envenomed rats. E. ocellatus venom elevated proinflammatory cytokines response as levels of tumor necrosis factor‐alpha and interleukin1‐beta significantly (P < 0.05) increased in cardiac tissues of the envenomed rats compared to control. The venom induced severe morphological defects in the heart tissues of envenomed rats indicating that E. ocellatus venom could actuate cardiotoxicity post envenoming.Item Inhibition of Echis ocellatus venom metalloprotease by flavonoid-rich ethyl acetate sub-fraction of Moringa oleifera (Lam.) leaves: in vitro and in silico approaches(Taylor & Francis, 2021) Adeyi, A.O.; Mustapha, K.K.; Ajisebiola, B.S.; Adeyi, O.E.; Metibemu, D.S.; Okonji, R.E"Envenoming by Echis ocellatus is potentially life-threatening due to severe hemorrhage, renal failure and capillary leakage. These effects are attributed to snake venom metalloproteinases (SVMPs). Due to drawbacks in the use of antivenin, natural inhibitors from plants are of interest in studies of new antivenin treatment. Antagonizing effects of bioactive compounds of Moringa oleifera is yet to be tested against SVMPs of E. ocellatus (SVMP-EO). Ethanol crude extract of M. oleifera was partitioned using n-hexane and ethyl acetate. Each partitions was fractionated using column chromatography and tested against SVMP-EO purified through ion exchange chromatography with EchiTab-PLUS polyvalent antivenin as control. Phytoconstituents of ethyl acetate fraction was screened against catalytic site of crystal of BaP1-SVMP while drug-likeness and ADMET toxicity of compound was equally determined. The molecular weight of isolated SVMP-EO was 43.28 kDa, with specific activity of 245 U/ml, percentage yield of 62.83 % and purification fold of 0.920. The Vmax and Km values are 2 mg/ml and 38.095 imol/ml/min, respectively while the optimal pH and temperature are 6.0 and 40 °C, respectively. Polyvalent antivenin, crude extract and ethyl acetate fraction of M. oleifera exhibited complete inhibitory effect against SVMP-EO activity. Inhibitions of P-1 and P-II metalloprotease's enzymes by ethyl acetate fraction are largely due to methanol, 6, 8, 9-trimethyl-4-(2-phenylethyl)-3-oxabicyclo[3.3.1]non-6-en-1-yl)- and paroxypropione, respectively. Both compounds are potential drug candidates with little or no concern of toxicity as revealed from in-silico predictions. The inhibitory effects suggest that this compound might be therapeutic candidate for further exploration for treatment of Ocellatus' envenoming."Item Moringa oleifera ethyl acetate fraction ameliorated the pathophysiological and histopathological alterations induced by Naja katiensis venom(Elsevier, 2021) Adeyi, A.O.; Ajisebiola, B.S.; Adeyi, O.E.; Badru, O.S.; Paul, O.; Idowu, B.BACKGROUND: Envenoming by Naja katiensis is known to constitute serious medical challenge. Moringa oleifera is a medicinal plant which have been reported as antivenom agent against pathologies caused by different snakes species. However, the ameliorative effects of extract and fractions of M. oleifera against toxicities induced by N. katiensis venom remain unreported. PURPOSE: This study therefore investigated the ameliorative effects of n-hexane and ethyl acetate fractions ob- tained from ethanol extract of M. oleifera leaves against N. katiensis venom-induced physiological and histological pathologies. METHODS: Histopathology and antiheamorrhagic activity of M. oleifera extract and fractions were studied using albino male Wistar rats while bovine citrated erythrocytes were used to determine the antiheamolytic and anti- coagulant effects. High Performance Liquid Chromatography was performed to chemically characterize the most active fraction of M. oleifera found to be most effective against venom-induced pathologies. RESULTS: N. katiensis venom induced severe cellular alterations in the brain, liver and kidney tissues of envenomed untreated rats. However, treatments with the fractions obtained from M. oleifera ethanol extract ameliorated tissue injuries in these organs and the most effective was ethyl acetate fraction. M. oleifera extract and fractions showed a dose dependent inhibitory effects on anticoagulant, hemolytic, and heamorrhagic activities of N. katiensis venom. The HPLC analysis of ethyl acetate fraction of M. oleifera leaf led to the identification of two major bioactive constituents as quercetin and kaempferol. CONCLUSION: This study proved that the ethyl acetate fraction of M. oleifera could serve as a potential source of effective compounds against N. katiensis venom-induced pathologies.