http://hdl.handle.net/123456789/5 Basic Medical Sciences 2026-04-15T14:11:16Z http://hdl.handle.net/123456789/2395 NEUROPROTECTIVE POTENTIALS OF THE ETHANOL EXTRACT OF Adenopus breviflorus (Benth) FRUIT IN SWISS MICE AND WISTAR RATS OYEBANJO, Oyetola Tolulope Neuroinflammation is a well-characterised feature of neurodegenerative diseases affecting individuals’ movement, memory and speech. The use of synthetic drugs to improve cognition and motor functions caused by inflammation in neurodegenerative diseases has been associated with adverse effects. Tropical fruits offer therapeutic approaches in mitigating inflammation peripherally and centrally. Adenopus breviflorus fruit used as anticonvulsant and pain reliever in folkloric medicine has not been fully evaluated for its neuroprotective activity. This study was designed to investigate the neuroprotective potentials of ethanol extract of Adenopus breviflorus (EEAB) fruit in Swiss mice and Wistar rats. Ripe fruits of Adenopus breviflorus were authenticated at Forest Herbarium Ibadan (FHI: 112244), air-dried and extracted with 70% ethanol to obtain EEAB. In the anti-nociceptive study, 30 male Swiss mice (25-30g) were divided into six groups (n=5): control (distilled water, 10mL/kg), EEAB (25, 50, 100, 200mg/kg) and indomethacin (10mg/kg). Anti- nociceptive activity was evaluated 1hour after by Acetic Acid-induced writhing Test (AAT). In the inflammation study, 30 male Wistar rats (150-180g), were divided into six groups (n=5): normal-saline (10mL/kg), carrageenan (1%, s.c.), EEAB (50, 100, 200mg/kg) and indomethacin (5mg/kg). Air-pouch was induced in the animals for six days after which animals in groups 3-6 were challenged with carrageenan for 24hours. Exudates from the air-pouch were evaluated for Total Leukocyte Counts (TLC) and neutrophils using standard techniques while Tumor Necrosis Factor-alpha (TNF-α) and Interleukin-6 were determined by ELISA. In the lipopolysaccharide-induced neuroinflammation model, 20 male Wistar rats (150-200g) were divided into four groups (n=5): normal saline (10mL/kg), lipopolysaccharide (250μg/kg, i.p.), EEAB (200mg/kg) and donepezil (0.5mg/kg i.p.). Memory function was assessed using Y-maze Tests (YMT). The Prefrontal Cortex (PFC), striatum and hippocampus were evaluated for inflammatory cytokines (TNF-α and Interleukin-6), neuronal morphology using Nissl and Golgi stains, and neuronal immunohistochemistry for expression of Amyloid-beta (Aβ) and Nuclear Factor kappa-B (NF-κB). Data were analysed using descriptive statistics and ANOVA at α0.05. vi The EEAB (100, 200mg/kg) significantly reduced writhing in AAT (27.83±8.89, 22.17±3.31) compared with control (46.67±4.03). The EEAB (100, 200mg/kg) relative to carrageenan-control significantly reduced TLC and neutrophil count. A significant reduction in TNF-α (337.20±29.29, 174.40±29.22 vs 521.80±35.02pg/ml exudate) and Interleukin-6 (289.90±26.48, 272.0±15.94 vs 392.10±17.69pg/ml exudate) of EEAB (100, 200mg/kg) compared with carrageenan-control was observed in the inflammation study. The EEAB (200mg/kg) significantly increased percentage alternations (69.72±2.56) in YMT relative to lipopolysaccharide-alone (55.63±2.82). The EEAB (200mg/kg) relative to lipopolysaccharide-alone significantly reduced TNF-α (pg/mg protein) in striatum (2.42±0.06 vs 3.19±0.12), and hippocampus (1.96±0.04 vs 2.32±0.10); and Interleukin-6 (pg/mg protein) in striatum (2.62±0.11 vs 3.71±0.33) alone. Neuronal morphology and dendritic arborization were preserved by EEAB (200mg/kg). The EEAB (200 mg/kg) relative to lipopolysaccharide-alone significantly decreased expressions of Aβ in the PFC (5.92±0.33 vs 15.46±0.77), and hippocampus (13.51±0.53 vs 17.19±1.44); and NF-κB in the PFC (8.61±1.03 vs 20.73±1.08), striatum (7.52±1.00 vs 18.84±0.77), and hippocampus (1.70±1.35 vs 14.08±1.13). Adenopus breviflorus fruit exhibited neuroprotective activities through repression of pro- inflammatory cytokines, amyloid beta and nuclear factor kappa-B in Swiss mice and Wistar rats. 2023-12-01T00:00:00Z http://hdl.handle.net/123456789/2393 EFFECTS OF INDOLE ACETIC ACID AND ULTRAVIOLET LIGHT ON SOME BIOCHEMICAL, NUTRITIONAL AND NEPHROPROTECTIVE PROPERTIES OF Moringa oleifera LAMARCK OMOLEKAN, Tolulope Omotope Chronic Kidney Diseases (CKD) are characterised by unresolved fibrosis causing progressive loss of kidney functions and Vitamin D Insufficiency (VDI). Humans synthesise Vitamin D (VD), an anti-fibrotic factor, when exposed to Growth Hormones (GH) and Ultraviolet B (UVB) light. However, VDI exacerbates fibrosis during Kidney Injury (KI). Many plants like Moringa oleifera (MO) synthesise VD in little amount, however, its enhancement using plant-GH {Indole Acetic Acid (IAA)} and exposure to UVB has not been sufficiently explored. This study was designed to assess IAA and UVB effects on MO’s biochemical, nutritional and nephro- protective properties. One hundred MO seeds (NH-41) were obtained from NIHORT, Ibadan. Twenty-five seeds were each soaked in distilled water (control), 0.4, 0.5, 0.6mM IAA and sowed in a randomised design (n=5) in polythene bags containing 10kg of soil. Seedlings, after seven days of germination, were exposed to UVB (315nm) at 1hour/day for 10weeks. The MO Leaves (MOL) were harvested, dried and milled, while MOL Oil (MOLO) was extracted with n-hexane using soxhlet apparatus. Coding sequences of MO lanosterol and cycloartenol synthases were amplified from cDNA of fresh MOL, cloned into plasmid, and transformed into yeast cells, respectively. The Transformed Yeast Cells (TYC) were treated with IAA (0.04mM) and exposed to UVB (15min).Vitamins D2 and D3 contents of MOL, IAA-UVB-treated-MOL, MOLO, IAA-UVB- treated-MOLO, and IAA-UVB-treated TYC were quantified by HPLC. Photosynthetic pigments, calcium and phosphorus contents of fresh MOL were determined spectrophotometrically. The MOL and MOLO were mixed with chow as experimental diets, separately. Twenty-five male mice (13-15g) were fed with adenine-fortified-chow (0.75%w/w) to initiate KI. Five mice without KI served as control. Mice with KI were grouped and fed with MOL (33%w/w), IAA-UVB-treated-MOL (33%w/w), MOLO (10%w/w), IAA-UVB-treated- MOLO (10%w/w), calcitriol-fortified chow (0.1%w/w) respectively, while control was fed with standard chow for four weeks, then sacrificed. Serum creatinine, Uric Acid (UA), sodium, phosphate and calcium were determined spectrophotometrically. Serum Fibroblast Growth Factor-23 (FGF-23), Kidney Injury Marker-1 (KIM1), and klotho were determined by ELISA. Kidney fibrosis was evaluated by H&E stains. Data were analysed using ANOVA at α0.05. Vitamins D2 and D3 contents of 0.6mM IAA-UVB-treated-MOL and IAA-UVB-treated-MOLO increased by 23.4, 25.3% and 29.2, 30.1%, respectively, relative to their controls. Vitamins D2 and D3 contents of IAA-UVB-treated TYC increased by 15.6, 11.7folds, respectively, relative to control. Chlorophylls a and b contents of 0.5mM IAA-UVB-treated-MOL (6.31±0.01 and 6.65±0.03mg/gfw), and 0.6mM IAA-UVB-treated-MOL (7.02±0.03 and 7.11±0.01mg/gfw), increased relative to control (4.90±0.01 and 4.99±0.01mg/gfw), respectively. Calcium and phosphorus contents of 0.5, 0.6mM IAA-UVB-treated-MOL increased by 44.6, 28.5% and 57.5, 59.1%, respectively, relative to control. The IAA-UVB-treated-MOLO reduced serum creatinine, UA, sodium, phosphate, calcium, FGF-23 and KIM1 by 25.0, 45.5, 25.0, 23.0, 26.5, 43.6 and 47.0%, respectively, relative to control. In IAA-UVB-treated-MOLO-fed mice, serum klotho increased by 45.0% and kidney fibrosis reduced significantly, relative to control. Indole acetic acid and ultraviolet B light enhanced vitamin D contents of Moringa oleifera leaves and its oil, which demonstrated nephro-protective properties via anti-fibrotic mechanisms. 2023-09-01T00:00:00Z http://hdl.handle.net/123456789/2340 ERECTOGENIC ACTIVITIES AND MECHANISM OF ACTION OF AQUEOUS EXTRACT OF Ocimum gratissimum LEAF AND THYMOL ON PENILE TISSUE IN WISTAR RATS SHITTU, Seyyid Alli-Sisse Erectile dysfunction is an increasing social and health concern. Relaxation of Corpus Cavernosum Smooth Muscle (CCSM) is required for penile erection. Thymol and thymol-containing plant, Ocimum gratissimum (OG), have been shown to have relaxant activity on aortic smooth muscle. However, modulation of smooth muscle relaxation is different across various tissues. There is thus, dearth of information on the erectogenic activities of OG or thymol. This study was designed to investigate the penile erectogenic activities of OG and thymol, and their probable mechanism of action in Wistar rats. Fresh OG leaves were procured from Bodija market, authenticated at Forestry Research Institute of Nigeria (FHI:110026), air-dried and macerated in distilled water to obtain Aqueous extract of OG (AeOG) which was lyophilised. Thirty-two male rats (170-190g) were used for in-vivo and ex-vivo experiments. Twenty rats for in-vivo study were grouped into four (n=5) and administered 0.5 mL/kg distilled water (control), 0.5 mL/kg corn oil, 300 mg/kg AeOG and 7.5 mg/kg analytical grade thymol orally for 28 days. Mounting latency and frequency were assessed as indices of mating behaviour. Serum Luteinizing Hormone and penile calcium-ATPase activities, Nitric Oxide (NO), cyclic Guanosine Mono-Phosphate (cGMP) and calcium levels were assayed spectrophotometrically. Ex-vivo effects of AeOG and thymol were assessed in 24 CCSM obtained from 12 rats. Using organ bath set-up, CCSM (n=6) were incubated in Kreb’s solution (control) or Kreb’s solution containing 60 μg/mL AeOG, 1% v/v ethanol or 0.06 μg/mL thymol. The CCSM were pre-contracted with phenylephrine (10-7M) or potassium chloride (KCl, 60 mM) for Maximum Contraction Response (MCR), and then relaxed with acetylcholine (10-5M) for Maximum Relaxation Response (MRR). Erectogenic mechanisms were then assessed via co-incubation with pathway inhibitors including Methylene Blue (MB, 10-4M), nifedipine (10-4M) or 5-(1,4-diazepan-1- ylsulfonyl)isoquinoline (HA-1077, 10-3M) for cGMP, calcium channel or Rho-Kinase blockade, respectively. Data were analysed using descriptive and ANOVA at α0.05. Mounting latency reduced in AeOG group by 59.6%. Mounting frequency increased by 63.8% and 47.7% in AeOG and thymol groups, respectively. Luteinizing Hormone (2.4±0.5 vs 1.1±0.1 μIU/mL) and Calcium-ATPase (4.3±0.1 vs 3.8±0.1 nmol/mg protein/hr) increased in AeOG group compared with control. Calcium (AeOG=18.7±0.3 and Thymol=18.8±0.2 10-4M/mL), NO (AeOG=11.4±2.0 and Thymol=12.9±1.6 μM/mL) and cGMP (AeOG=32.3±1.3 and Thymol=33.2±1.1 pM/mL) levels were similar to control (18.8±0.2 10-4M/mL, 11.6±1.1 μM/mL, 33.1±0.6 pM/mL, respectively). The MCR to phenylephrine (77.5±2.2%) was reduced by AeOG (67.2±4.6%) and thymol (68.7±1.1%), while MRR was only increased by thymol (65.0±1.9%) compared with control (54.2±2.1%). The MRR in AeOG (52.6±3.74%) was promoted by MB (60.7±3.4%) and reduced by nifedipine (46.1±1.9%). None of the inhibitors reduced the MRR in thymol. During KCl pre-contraction, MCR was reduced by AeOG (51.5±3.1%) and thymol (57.6±1.5%) compared with control (63.1±2.9%). Both AeOG (64.4±2.6%) and thymol (54.7±2.6%) increased MRR compared with control (47.6±3.4%). The effect of AeOG and thymol on MRR were reduced by nifedipine (56.8±2.9% and 47.5±2.3%, respectively) and HA-1077 (52.2±1.6% and 45.5±2.4%, respectively). The aqueous extract of Ocimum gratissimum leaf and thymol promoted penile erection. This erectogenic activity involved reduction of penile smooth muscle contraction sensitivity to calcium via Rho-Kinase pathway. 2022-02-01T00:00:00Z http://hdl.handle.net/123456789/2266 ISOLATION OF STIGMASTEROL FROM Piptadeniastrum africanum (HOOK. F.) AND ITS MODULATORY EFFECT ON MITOCHONDRIAL-MEDIATED CELL DEATH IN LIVER AND COLONIC TOXICITY IN MICE OLOJO, Folake Olayinka Hepatocellular and colonic damage are fatal outcomes arising from liver and colon toxicity. Modulation of mitochondrial-mediated cell death is a strategy in these diseases management. The use of synthetic drugs in the treatment of these diseases presents several side effects. In folklore, Piptadeniastrum africanum (PA) is used for the treatment of these disorders but the mechanism has not been explored. This study was designed to investigate the role of bioactive compound(s) purified from PA on liver and colon damage via the modulation of mitochondrial-dependent cell death. The istem ibark iof iPiptadeniastrum iafricanum iwas icollected ifrom ia iforest iat iIbadan, identified iat ithe idepartment iof iBotany, iUniversity iof iIbadan i(UIH-22562) iair-dried, pulverised iand isoaked iin iabsolute imethanol ito iobtain iits iextract i(CMEPA). i iThe iCMEPA was ifractionated iusing in-hexane, ichloroform, iethyl iacetate iand imethanol ito iobtain itheir respective ifractions i(HFPA, iCFPA, iEFPA iand iMFPA) iwhich iwere itested ion imitochondrial permeability itransition i(mPT) ipore iopening iand iATPase iactivity i(in ivitro). i iThe ieffects iof the ifractions iof iPA ion iliver iand icolon itissues iwere ievaluated iin istudy i1. iTwenty imice (20±2g; in=5) iwere igrouped iand itreated ias ifollows, igroups i1 i(vehicle), i2, i3 iand i4 iwere treated iintraperitoneally iwith i25, i50 iand i100 img/kg iof iEFPA ionce idaily ifor i14 idays iand liver imPT iassayed. iThe iEFPA iwas ipurified iusing icolumn ichromatography ito iobtain ia ipure compound iwhich iwas icharacterised ias istigmasterol iby ispectroscopic itechniques. iIn istudy i2, forty-two imice iwere igrouped i(n=7), itreated iwith iDextran iSulphate iSodium i(DSS) iand Benzo{a}Pyrene i(BaP) ifor i10 days as follows, groups 1 (vehicle), 2 (oral 4% DSS), 3 (125mg/kg BaP), 4 (DSS+BaP), 5 (DSS+BaP and 200mg/kg of stigmasterol) and 6 (DSS+BaP and 400mg/kg of stigmasterol). The mice were sacrificed and colon samples prepared. Tumor Necrosis Factor-α (TNF-α), Interleukin 6 (IL-6), p53, Caspases 9 (C9), Bax, Bcl-2, were performed on the colon using immunohistochemistry. All data were analysed using descriptive statistics and ANOVA at α0.05. In vitro, EFPA had the highest induction of mPT pore opening (3.80, 5.60, 6.40, 8.10 and 8.90 folds) at 20, 60, 100, 140 and 180µg/ml, respectively and enhanced ATPase activity (0.20±0.01, 0.35±0.10, 0.40±0.10, 0.45±0.20 and 5.20±0.80µmole/Pi/mg/protein/min), relative to the vehicle (0.05µmole/Pi/mg/protein/min), respectively. In vivo, EFPA caused mPT induction of 2.50, 4.90 and 6.90 folds at 25, 50 and 100mg/kg, respectively. The toxicant groups (DSS, BaP and DSS+BaP), relative to the vehicle significantly increased TNF-α (40.00±1.20%, 30.00±0.90%, 34.00±1.10% vs 15.00±0.70%), IL-6 (160.00±3.50%, 110.00±2.20%, 120.00±1.50% vs 60.00±1.50%) and p53 (80.00±2.30%, 70.00±1.10%, 85.00±2.20% vs 25.00±0.90%) However, stigmasterol (200 and 400mg/kg), relative to DSS+BaP significantly attenuated TNF-α (12.00±0.09%, 13.00±0.10% vs 34.00±1.10%), IL-6 (53.00±1.30%, 50.00±1.20% vs 120.00±1.50%), p53 (40.00±2.20%, 50.00±1.70% vs 85.00±2.20%). The DSS, BaP and DSS+BaP, relative to the vehicle increased C9 (1.10±0.10, 0.90±0.01, 1.10±0.10ng/ml vs 0.01ng/ml), Bax (120.00%, 100.00%, 90.00% vs 110.00%) and reduced Bcl-2 (80.00%, 75.00%, 75.00% vs 90.00%) which were modulated by stigmasterol. Stigmasterol isolated from the Piptadeniastrum africanum modulated mitochondrial-mediated cell death via a decrease in pro-inflammatory cytokines and levels of pro-apoptotic proteins. 2023-04-01T00:00:00Z