Basic Medical Sciences
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Browsing Basic Medical Sciences by Advisor "Tiloke, Charlette"
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Item Open Access The hepatoprotective effects of ๐๐ฐ๐ณ๐ช๐ฏ๐จ๐ข ๐ฐ๐ญ๐ฆ๐ช๐ง๐ฆ๐ณ๐ข against antiretroviral-induced cytotoxicity in HepGโ cells(University of the Free State, 2023) Saki, Mbasakazi; Tiloke, Charlette; Ntsapi, Claudia; De Villiers, Helena๐๐ป๐๐ฟ๐ผ๐ฑ๐๐ฐ๐๐ถ๐ผ๐ป: The untreated human immunodeficiency virus (HIV), a lentivirus species that attacks immune cells, causes acquired immunodeficiency syndrome (AIDS). HIV/AIDS is managed by Antiretroviral therapy (ART). The ART regimen contains nucleoside reverse transcriptase inhibitors (NRTIs) associated with oxidative stress. Medicinal plants are often combined with ART to diminish the side effects of ART use. The ๐๐ฐ๐ณ๐ช๐ฏ๐จ๐ข ๐ฐ๐ญ๐ฆ๐ช๐ง๐ฆ๐ณ๐ข (MO) tree extracts have been shown to contain bioactive compounds with antioxidant effects. ๐๐ถ๐บ: This ๐ช๐ฏ ๐ท๐ช๐ต๐ณ๐ฐ study evaluated the cytotoxicity of an NRTI (tenofovir) and its potential amelioration by MO leaf extract. ๐ ๐ฒ๐๐ต๐ผ๐ฑ๐: HepGโ cells were exposed to tenofovir, MO, and combination (tenofovir and MO) treatment groups for 24 and 120 hours. MO aqueous leaf extract was prepared, and cytotoxicity was assessed. Markers for oxidative stress and antioxidant response were assessed using spectrophotometry, luminometry, ELISA, qPCR, and western blotting experimental techniques. ๐ฅ๐ฒ๐๐๐น๐๐: At 24 hours, tenofovir decreased MDA ๐๐๐2, ๐๐๐2, ๐๐๐ mRNA, and NRF2, SOD2, and CAT protein expression. It then increased GSH, ๐๐๐๐ mRNA and p-NRF2 protein expression. MO decreased GSH levels, NRF2, ๐๐๐๐, and ๐๐๐2 mRNA expression and increased ๐๐๐ mRNA, as well as NRF2, p-NRF2, SOD2, and CAT protein expression. At 120 hours, tenofovir increased MDA, NRF2 mRNA, NRF2, p-NRF2, and SOD2 protein expression. It then decreased GSH levels, ๐๐๐๐, ๐๐๐2, ๐๐๐ mRNA and CAT protein expression. MO decreased MDA and GSH levels, NRF2 and CAT protein expression. It then increased ๐๐๐2, ๐๐๐๐, ๐๐๐2, ๐๐๐ mRNA, p-NRF2, and SOD2 protein expression. The combination treatment group downregulated MDA and upregulated the expression of NRF2, ๐๐๐๐, ๐๐๐2, ๐๐๐ mRNA and NRF2, p-NRF2, SOD2, and CAT proteins. ๐๐ผ๐ป๐ฐ๐น๐๐๐ถ๐ผ๐ป: Adding MO to tenofovir downregulates reactive oxygen species by upregulating the NRF2-antioxidant pathway to reduce oxidative stress. Therefore, MO has the potential to ameliorate toxicity induced by tenofovir.Item Open Access Investigating the potential antiproliferative effect of ๐๐ฐ๐ณ๐ช๐ฏ๐จ๐ข ๐ฐ๐ญ๐ฆ๐ช๐ง๐ฆ๐ณ๐ข aqueous leaf extract in MCF-7 breast cancer cells(University of the Free State, 2023) Moremane, Malebogo M.; Tiloke, Charlette; Abrahams, Beynon๐๐ฎ๐ฐ๐ธ๐ด๐ฟ๐ผ๐๐ป๐ฑ: Breast cancer is associated with elevated mortality and morbidity rates in women across the world. Current chemotherapeutic drugs such as Doxorubicin (Dox) display contra-indications, thus expressing the need for alternative treatment methods. Therefore, to reduce the cancer burden, the studyโs objective was to investigate whether an aqueous leaf extract of ๐๐ฐ๐ณ๐ช๐ฏ๐จ๐ข ๐ฐ๐ญ๐ฆ๐ช๐ง๐ฆ๐ณ๐ข (MO), a medicinal tree native to India and indigenous to Africa, possesses antiproliferative potential against MCF-7 breast cancer cells. ๐ ๐ฒ๐๐ต๐ผ๐ฑ๐ผ๐น๐ผ๐ด๐: In order to suppress cell growth, MCF-7 cells were treated with MO (2600 ฮผg/ml) for 72 hours. Cells were also co-exposed with Dox (0.978 ฮผM), modelled as a positive control. The unexposed cells served as the control. Biochemical analysis was conducted after 72 hours (MTT, GSH, DCFH-DA, ATP, Caspase 3/7, 8/9, qPCR and western blot assays) to assess the efficacy of MO and Dox. ๐ฅ๐ฒ๐๐๐น๐๐: ๐๐ฐ๐ณ๐ช๐ฏ๐จ๐ข ๐ฐ๐ญ๐ฆ๐ช๐ง๐ฆ๐ณ๐ข aqueous leaf extract significantly reduced the proliferation of breast cancer cells by inducing oxidative stress through increasing ROS whilst decreasing glutathione content and Nrf2 protein expression. Additionally, MO induced apoptosis by increasing caspases -3/7, -8, -9, metabolic activity and upregulating p53. Similar results were observed in Dox-exposed cells. Furthermore, cell death due to MO was activated with downregulation of Bcl-2, PARP-1 and Bax. Dox decreased the growth of breast cancer cells by increasing ROS. In contrast, Dox induced chemoresistance through increased GSH content and downregulated apoptotic protein Bax and p53 gene. However, the MO + Dox combination induced antiproliferative potential similarly to MO, suggesting a possible synergistic effect. ๐๐ผ๐ป๐ฐ๐น๐๐๐ถ๐ผ๐ป: MO aqueous leaf extract displayed antiproliferative potential by inducing apoptosis and oxidative damage to the MCF-7 breast cancer cells