http://hdl.handle.net/123456789/59 2026-04-13T10:38:43Z 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/123456789/2340 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/123456789/1973 SEXUAL DIMORPHIC RESPONSE OF METABOLIC VARIABLES IN HIGH FAT DIET-INDUCED OBESITY IN WISTAR RATS ON LOW CALORIE DIET DAVID, Ubong Edem Striking differences exist between men and women in lipid kinetics possibly due to sexual dimorphism in metabolism. However, the effect of prolonged intake of High Animal Fat Diets (HAFD), High Plant Fat Diets (HPFD) and Low Calorie Diets (LCD) in both sexes have not been fully investigated. This study was designed to evaluate the influence of sexual dimorphism on metabolic variables following prolonged intake of HPFD and HAFD in normal, and LCD-treated obese Wistar Rats (WR). Gross energy, fat and fiber content of formulated feeds from proximate composition were HPFD (4.50 Kcal/g, 16.30, 3.60 %), HAFD (5.90 Kcal/g, 31.00, 3.10 %) and LCD (2.66 Kcal/g, 2.27, 20.64%). In experiment one (designed to evaluate sexual dimorphism in high fat diets induced obesity rats), 30 WR were divided into 15 males (mWR) and 15 females (fWR). Each sex group was sub-divided into 3 groups (n=5) and fed with Standard Chow (SC), HPFD and HAFD respectively, for 17 weeks. Serum was obtained from blood, samples of the liver, Small Intestine (SI) and heart were excised. Serum, liver, heart and SI lipid profile [High Density Lipoprotein (HDL), Low Density Lipoprotein (LDL), Total Cholesterol (TC), triglyceride, Free Fatty Acids (FFA)] were measured by spectrophotometry. Serum luteinizing hormone (LH), Apolipoprotein A and B were analysed using ELISA while SI Saturated, Monounsaturated and Polyunsaturated Fatty Acids (SFA, MUFA and PUFA, respectively) were measured by gas chromatography using standard methods. Duodenal section of SI was examined for Cluster of Differentiation 36 (CD36) expression using immunohistochemistry. In experiment two, 30 WR were divided into two sexes (n=15). Each sex subgroup was divided into 3 groups (n=5). Subgroups 1 and 2 were fed on SC and HPFD for 22 weeks while subgroup 3 was fed HPFD for 17 weeks followed by LCD for 5 weeks. The same experimental analyses in study one were thereafter implemented. Data were analysed using descriptive statistics and ANOVA at α0.05. In experiment one, the SC showed no significant difference in LDL, TC and triglyceride between fWR and mWR. In HPFD; serum FFA, heart LDL, SI SFA decreased significantly while heart HDL increased significantly in fWR compared with mWR (815±55.00 vs 1754±75.00µmol/L; 27±1.80 vs 38±2.90mg/dL; 41.03±2.07 vs 50.49±1.31%; 20±1.50 vs 14±1.40mg/dL, respectively). The HAFD significantly increased serum TC, LH, apolipoprotein B:A ratio, SI PUFA in fWR compared with mWR (251±17.00 vs 191±3.90mg/dL; 3.5±0.09 vs 4±0.20µlU/mL; 3.5±0.46 vs 2.8±0.07; 13.47±1.34 vs 8.5±0.75%, respectively). No significant difference was observed for MUFA between the sexes. Duodenal section of SI showed increased CD36 expression in fWR compared with mWR (5.83±0.26 vs 8.65±0.83%). In experiment two, serum TC and triglyceride significantly increased in subgroup 2, while in subgroup 3, liver LDL, TG and FFA significantly reduced in fWR compared with mWR (258±11.00 vs 217±9.80; 272±19.00 vs 202±6.90; 14±0.85 vs 31±1.60; 109±2.1 vs 182±5.8mg/dL; 834±72 vs 1409±74µmol/L respectively). Sexual dimorphic response that was more pronounced in males on low calorie diet may be associated with sexually distinct lipid profile and hormonal modulations between the sexes. 2023-02-01T00:00:00Z http://hdl.handle.net/123456789/1486 ANTIHYPERTENSIVE POTENTIAL OF AQUEOUS EXTRACT OF Peristrophe bivalvis (L.) MERR. LEAF ON NG-NITRO L-ARGININE METHYL ESTER-INDUCED HYPERTENSION IN MALE WISTAR RATS ALUKO, ESTHER OLUWASOLA Hypertension is a major risk factor for cardiovascular diseases. Most antihypertensive drugs have been reported to effectively reduce blood pressure (BP), but they do not ameliorate end-organ damage associated with hypertension. This necessitates the need for agents that can effectively manage hypertension and its complications. Peristrophe bivalvis (PB) is used in traditional medicine to treat hypertension and other ailments. This study was designed to investigate the effects of aqueous extract of PB leaf (APB) on NG-nitro-L-arginine methyl ester (L-NAME)-induced hypertension and its associated cardio-renal complications in male Wistar rats. Peristrophe bivalvis leaf was obtained in Ikono, Akwa-Ibom State, and authenticated at the Department of Pharmacognosy and Herbal Medicine, University of Uyo (ID: UUHO43). Aqueous extract of PB leaf was obtained using cold maceration, filtration and concentration. Twenty-five male Wistar rats (150-170 g) were grouped into five (n=5). Group 1 received 10 mL/kg of distilled water (control) while groups 2-5 were administered with 60 mg/kg of L-NAME (L-NAME60, hypertensive [H]) orally for eight weeks to induce hypertension. After eight weeks, groups 2-5 received: L-NAME60+distilled water (hypertensive untreated [HuT]), L-NAME60+APB (200 mg/kg, hypertensive APB [HAPB]), L-NAME60+ramipril (10 mg/kg, hypertensive ramipril [HRM]) and distilled water (hypertensive recovery [R]) respectively, for five weeks. The BP was mea¬sured by tail-cuff method at 8th and 13th weeks of the study. The rats were sacrificed and samples (blood, heart and kidney) collected. Serum angiotensin II, cyclic Guanosine Monophosphate (cGMP), tissue Interleukin-1 beta (IL-1β) and Tumor Necrosis Factor-alpha (TNF-α) levels were measured using ELISA technique. Tissue malondialdehyde, Superoxide Dismutase (SOD) and reduced Glutathione (GSH) levels were measured by spectrophotometry. Nitrotyrosine and Cluster of Differentiation 68 (CD68) expressions in the tissue were quantified using immunohistochemistry. Data were analysed using ANOVA at α0.05. Systolic, diastolic and mean arterial BP significantly decreased in HAPB (161±1.30, 117±2.60, 132±1.60 mmHg), HRM (132±1.10, 100±1.10, 110±0.86 mmHg) and R (152±2.90, 117±2.30, 128±2.40 mmHg) groups compared to H (172±2.80, 131±2.50, 144±2.50 mmHg) and HuT (196±2.0, 159±2.80, 168±4.90 mmHg) groups, respectively. Angiotensin II level significantly decreased while cGMP level increased in HAPB group compared to HuT (32.49±0.45 vs 51.34±1.02 pmol/mL, 9.96±0.53 vs 6.50±0.62 pmol/mL, respectively). Levels of IL-1β (pg/g tissue) and TNF-α (ng/g tissue) in the heart (3609.33±372.18, 121.03±6.0) and kidney (4635.78±62.58, 174.79±16.92) of HAPB group significantly decreased relative to HuT heart (5157.11±113.66, 296.21±3.16) and kidney (5122.04±207.90, 289.99±6.17), respectively. Administration of L-NAME60+APB significantly decreased malondialdehyde (µmol/g tissue) level and increased SOD (units/g tissue) activity and GSH (mmol/g tissue) level in the heart (17.05±0.40, 2.50±0.05, 1.10±0.08) and kidney (15.43±0.63, 2.60±0.05, 1.20±0.06) compared to HuT heart (69.49±5.43, 0.39±0.03, 0.37±0.06) and kidney (61.39±4.80, 0.41±0.03, 0.44±0.07), respectively. Nitrotyrosine and CD68 expressions were decreased in the heart and kidney of HAPB group relative to HuT group. Blood pressure level significantly increased but nitrotyrosine expression in the heart decreased in HAPB group compared to HRM. 2021-09-01T00:00:00Z