http://hdl.handle.net/123456789/54 2026-04-18T14:41:34Z 2026-04-18T14:41:34Z OMOLEKAN, Tolulope Omotope http://hdl.handle.net/123456789/2393 2025-11-06T15:09:22Z 2023-09-01T00:00:00Z

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 OLOJO, Folake Olayinka http://hdl.handle.net/123456789/2266 2024-05-23T08:28:53Z 2023-04-01T00:00:00Z

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 SAMUEL, EKUNDAYO STEPHEN http://hdl.handle.net/123456789/1963 2024-04-24T16:37:51Z 2023-02-01T00:00:00Z

CYTOPROTECTION BY LEAF EXTRACT OF Tridax procumbens (LINN.) IN EMBRYONIC MOUSE, HEPG2 AND PANC-1 CELL LINES AND ARSENITEINDUCED TOXICITY IN RATS SAMUEL, EKUNDAYO STEPHEN Pancreatic Ductal Adenocarcinoma (PDA) and Hepatocellular Carcinoma (HCC) are among the deadliest cancer types worldwide that are associated with arsenic intoxication. Current drugs used in cancer management show a lot of side effects. There is, therefore, an increased search for alternatives from medicinal plants with anticancer properties. Tridax procumbens (TP) is a medicinal plant rich in antioxidant phytochemicals with little information on its effects on cancer and arsenic toxicity. This study was designed to investigate the effect of Tridax procumbens in PDA, HCC and arsenite-induced toxicity using in vivo and in vitro models. The TP was authenticated (UIH-22542), air-dried, blended, soaked in ethanol, and concentrated to obtain the Crude Extract (CETP). The CETP was fractionated using Hexane, Dichloromethane, and Ethyl Acetate to obtain Fractions (HXF, DCMF, and EAF, respectively). Antioxidant assays of TP were performed. For in vivo study, 32 male Wistar rats (80-100g) were assigned into four groups (n=8), and treated as follows; A (control); 1 ml/kg body weight olive oil, B; 2.5 mg/kg Sodium Arsenite (SA), C; 50 mg/kg CETP, D; SA+CETP. Olive oil and CETP were administered once daily for 14 days, and SA twice (days 7 and 14). Histological examination of lungs and brain, and micronucleated Polychromatic Erythrocytes (mPCEs) frequency were carried out. The HCC and PDA cell lines; HepG2 and Panc-1, were maintained in a humidified incubator, and treated with (10, 20, 50, 100, and 250 𝜇g/ml) dimethyl sulfoxide (control), CETP and CETP-fractions for 24 and 48 hours. Similarly, embryonic mouse pancreas (E11.5d) were cultured for five days and treated with the test samples (20 𝜇g/ml) for two days. Cytotoxicity assays [3- (4,5-dimethylyhiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), Live/Dead] were carried out, and expression of proteins (cytokeratin-7, peanut agglutinin (PNA), α- fetoprotein, insulin, glucagon, amylase, catalase, alkaline phosphatase, Glutathione Stransferase-pi (GST-pi), caspase-3, Bcl-2, Adenomatous Polyposis Coli (APC), p53, p21Cip1/Wap-1, vimentin, Ki-67, Sox9, β-catenin, α1-antitrypsin, albumin, transferrin) were evaluated using immunofluorescence. Data were analysed using descriptive statistics and ANOVA at α0.05. The DCMF scavenged 2,2-diphenyl-1-picrylhydrazyl hydrate and nitric oxide radicals (IC50: 0.43 and 0.39 𝜇g/ml, respectively) relative to control (IC50: 0.85 and 0.14 𝜇g/ml, respectively). The CETP reduced arsenite-induced histological lesions in lungs and brain, and mPCEs [SA (21.00±3.33) to SA+CETP (3.80±0.58)] compared to control (0.60±0.40). The DCMF elicited cell death (IC50=23.1 𝜇g/ml) compared to CETP (IC50=114.2 𝜇g/ml). Relative to control, phenotype morphogenesis was observed in DCMF-treated embryonic pancreas with immunopositivity for insulin, vimentin and amylase (1.3-fold), cytokeratin- 7 (1.5-fold), PNA (1.9-fold), and glucagon (2.8-fold). There were significant elevations in transferrin (1.5-fold), albumin (1.8-fold), p53 (2.8-fold), caspase-3 (2.9-fold), catalase (4.0-fold), p21Cip1/Wap-1 (4.4-fold), and alkaline phosphatase (5.0-fold) in DCMF-treated cells compared to control. Conversely, there were significant reductions in β-catenin (1.3- fold), α1-antitrypsin and cytokeratin-7 (2.0-fold), PNA and GST-pi (2.3-fold), α- fetoprotein and Ki-67 (2.7-fold), and vimentin (10.8-fold) in DCMF-treated cells relative to control. The DCMF induced Bcl-2 punctate nuclear staining, and cytoplasmic translocation of APC, Sox9, and β-catenin in treated cells. Leaf extract of Tridax procumbens exhibited antiproliferative activities via the decrease in biomarkers of cancer induction and increase cellular antioxidant defence system.

2023-02-01T00:00:00Z ADEFISAN, ADEDOYIN OMOBOLANLE http://hdl.handle.net/123456789/1961 2024-04-24T16:34:20Z 2023-02-01T00:00:00Z

CHEMOPREVENTIVE EFFECTS OF ROOT BARK EXTRACT OF C. PORTORICENSIS (BENTH.) IN N-NITROSO-N-METHYLUREA AND BENZO(A)PYRENE-INDUCED MAMMARY GLAND AND REPRODUCTIVE TOXICITY IN WISTAR RATS ADEFISAN, ADEDOYIN OMOBOLANLE Breast cancer (BC) remains a disease with high morbidity and mortality. In Nigeria, BC represents about 23% of all cancer cases. Currently available chemotherapeutic interventions are associated with significant adverse effects, hence the need for safer options. C. portoricensis (CP) has been applied to manage breast diseases in ethnomedicine. However, there is paucity of scientific basis to justify this claim. In this study, an investigation of the effects of CP on N-nitroso-N-methylurea (MNU) and Benzo(a)pyrene (BP)-induced mammary and reproductive organ toxicity in Wistar rats was conducted. A root bark sample of CP was taken from Ikire, Osun State and authenticated at Forest Hebarium Ibadan (FHI:111949). The methanol extract of CP (MCP) was obtained from the powdered root. The MCP was partitioned to obtain n-hexane, chloroform, ethylacetate and butanol fractions. The chloroform fraction of CP (CCP) was subjected to biochemical assay using MCF-7 cells. Sixy-four female Wistar rats (30-40g) were divided into eight groups (n=8): Vehincle, MNU, [MNU+MCP (100 mg/kg)], [MNU+MCP (200 mg/kg)], [MNU+MCP (300 mg/kg)], MCP (300 mg/kg), [MNU+Vincasar (0.5 mg/kg)] and Vincasar only. In another study, fifty-six female rats were grouped into seven (n=8): Vehincle, [MNU+BP], [MNU+BP+CCP (50 mg/kg)], [MNU+BP+CCP (100 mg/kg)], CCP (100 mg/kg), [MNU+BP+ Vincasar] and Vincasar only. The MNU (50 mg/kg) and BP (50 mg/kg) were administered intraperitoneally at age 7, 10 and 13 weeks. The MCP and CCP were administered orally thrice weekly. Rats were sacrificed; blood and tissues (mammary and uterine) were obtained for analyses. Lactate dehydrogenase (LDH), Superoxide Dismutase (SOD), Catalase, Glutathione-S-Transferase (GST), Glutathione Peroxidase (GPx), malondialdehyde, Nitric Oxide (NO) and Myeloperoxidase were determined by visspectrophotometry. Cyclooxygenase-2, inducible Nitric Oxide Synthase (iNOS), B-cell lymphoma-2 (Bcl-2), p53, Interleukins-1β and 6 (IL-1β, IL-6), Estrogen and Progesterone receptors (ER+, PR+) and Epidermal Growth Factor Receptor-2 (EGFR- 2) were determined by immunohistochemistry. Micro-section of tissues were subjected to Hematoxylin & Eosin and examined under the light microscope. Data were analysed using ANOVA at α0.05.ix The CCP decreased levels of IL-1β (34%), LDH (24%), MPO (74%), malondialdehyde (55%) and increased the activities of SOD (48%) and catalase (49%) in MCF-7 cells. The MNU decreased activities of mammary SOD (22%), uterine CAT (24%) and, GST (25%) relative to Vehincle. The MCP (100 mg/kg) when compared with MNU significantly elevated mammary SOD (18%) and uterine GST (20%). Treatment with CCP (100 mg/kg) when compared with MNU+BP significantly increased the mammary catalase (24.42±4.86 vs 16.1p6±2.90), GPx (171.48±13.97 vs 93.68±5.06), SOD (25.16±4.34 vs 16.09±2.90) and uterine catalase (29.52±4.83 vs 15.04±2.41) and SOD (48.56±4.70 vs 21.42±0.35), respectively. Additionally, CCP (100 mg/kg) significantly decreased mammary and uterine MPO (73%, 57%), NO (21%, 26%) and malondialdehyde (10%, 31%) when compared with [MNU+BP]-treated rats. The CCP (100 mg/kg) decreased ER+, PR+, EGFR-2, cyclooxygenase-2, iNOS, IL-6, IL-1β, Bcl- 2 and increased p53 expression in the mammary tissues of [MNU+BP+CCP]-treated rats. Histology of mammary tissues showed atypical epithelia, high nucleocytoplasm and ductal carcinoma in MNU+BP rats, which were reversed in rats given CCP (100 mg/kg). C. portoricensis demonstrated chemopreventive effects via induction of apoptosis and reduction of inflammatory cytokines.

2023-02-01T00:00:00Z