Browsing by Author "Ngcobo, Mlungisi."

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The biochemical effects of Sutherlandia Frutescens in cultured H9 cancerous T cells and normal human T lymphocytes.
(2008) Ngcobo, Mlungisi.; Chuturgoon, Anil Amichund.
Indigenous plants have long been used by African populations in their cultural lives and health care. Sutherlandia frutescens (SF) is a popular traditional medicinal plant found in various parts of southern Africa and used for treatment or management of different diseases, including cancer and HIV/AIDS. In this study, the biochemical effects of various dilutions (1/50, 1/150, 1/200, and 1/300) of SF 70% ethanol (SFE) and deionised water (SFW) extracts in cancerous H9 and normal T cells were examined. Untreated, 70% ethanol-treated and camptothecin (CPT, 20jiiM) treated cells were used as reference samples for comparison. Cytotoxicity, apoptotic enzymes activity, oxidant scavenging and antioxidant promoting abilities, cellular morphology and cytokine signalling effects were assessed using the methylthiazol tetrazolium (MTT) assay, adenosine triphosphate (ATP) assay, caspase-3/-7 activity assay, thiobarbituric acid reactant substance (TBARS) and glutathione (GSH) assays, fluorescence microscopy and an ELISAbased cytokine analyses assay respectively. Sutherlandia frutescens ethanol and water extract dilutions (1/50 and 1/200) were shown to be cytotoxic to H9 T cells in a dose- and time-dependent manner with the SFE extract having an average IC50 of 1/40 after 24 hours while SFW extract reached a similar IC50 only after 48 hours. In normal T cells, the SFE extract induced proliferation after 24 hours but this was reverse after 48 hours. The SFW extract dilutions did not significantly change cell viability after 24 hours but significantly increased cell viability after 48 hours. Both SFE and SFW extracts dilutions induced a dose- and time-dependent inhibition of caspase-3/-7 activity in both H9 and normal T cells. Both types of extracts were also shown to efficiently remove lipid peroxides from supernatants of treated cell lines, with SFW extract having a more lasting effect. In the GSH assay, the SFE and SFW extract dilutions reduced GSH levels in H9 T cells, with the SFW extract dilutions being more effective. In normal T cells, the higher dilutions (1/150 and 1/300) of SFW extract increased GSH levels significantly while lower dilutions (1/50) of both SFE and SFW extracts significantly inhibited GSH levels. Lower dilutions (1/50) of SFE and SFW extracts induced chromatin condensation in both H9 and normal T cells after 48 hours incubation. Using treated peripheral blood mononuclear cells (PBMCs) supernatants, SFE and SFW extract dilutions were shown to reduce the levels of pro-inflammatory cytokines IL 1 p and TNF-a in a dose-dependent manner. These results further confirmed the anticancer abilities of SF and showed that higher concentrations of this medicinal plant can be toxic to normal T cells in vitro while lower concentrations can stimulate the immune cells. Therefore further studies should be conducted with regards to the effects of SF on the immune system in both in vitro and in vivo systems.
Concurrent treatment of in vitro cell lines with uthuli lwezichwe™, an African traditional medicine used in the management of diabetes mellitus in KwaZulu-Natal, with conventional treatments and their effects on glucose uptake and insulin secretion.
(2022) Hlatshwayo, Sphamandla.; Ngcobo, Mlungisi.; Gqaleni, Nceba.
Background: By the year 2025, prevalence of diabetes mellitus is estimated to reach 300 million globally; with type 2 diabetes mellitus comprising more than 90% of these cases. Due to the side effects which are sometimes adverse and gradual loss of efficacy with time, presented by conventional regimens; a number of diabetic patients have been reported to be using both traditional medicines and conventional regimens concurrently. Experimental and clinical experiments have yielded positive results on studies performed on conventional treatments in combination with traditional medicines and medicinal plant extracts. Most these studies have been performed using a single medical plant, whilst African traditional medicine (ATM) products constitute of a variety of medical plants. Aim: This study aimed to investigate the concurrent treatment of in vitro cell lines with Uthuli Lwezichwe™, an African traditional medicine used in the management of diabetes mellitus in KwaZulu-Natal, in combination with conventional treatments and their effects on glucose uptake and insulin secretion. Methods: Cell viability was used to establish the IC50 doses of Uthuli Lwezichwe™ for HepG2 liver, C2C12 muscle and RIN-5 pancreatic beta cell lines. The IC50 doses were used in combination with known effective doses of metformin, insulin and tolbutamide to treat liver, skeletal muscle and beta cells, respectively. Glucose uptake was monitored at 0, 6, 12 and 24 h time intervals. Changes in glycogen and glutathione (GSH) levels in treated liver cells were evaluated using a glycogen assay kit (MAK0160) and GSH Glo glutathione kits, respectively. Insulin secretion in treated pancreatic cells was assessed using an ultra-sensitive rat insulin ELISA kit. Results: In comparison to the untreated control, treatment with Uthuli LwezichweTM in combination with conventional drugs significantly increased (p<0.05) increased insulin secretion in comparison to all treatment groups. Conclusion: Interaction of anti-diabetic agents studied, resulted in ameliorated glucose metabolism, both via glucose uptake and insulin secretion. This could be beneficial both in modulating diabetes mellitus and its comorbidities.
Immunomodulatory and anticancer potential of a traditional medicine product from a traditional healer against MCF-7 breast cancer cells.
(2021) Sithole, Siboniso Percival.; Ngcobo, Mlungisi.
The incidence of breast cancer in South Africa is expected to rise dramatically in the forthcoming decade. In 2018, approximately 2 088 849 new cases and 626 679 cases of deaths from breast cancer alone were reported worldwide. Among African countries, the incidence of cancer is higher in southern Africa with an estimate standard rate of 235.9 per 1000, 0000 people. Currently breast cancer is treated with either chemotherapy, surgery, radiotherapy or immunotherapy. All this conventional treatment options available for breast cancer have some limitations and drawbacks. Therefore, there is an urgent need for the development of novel breast cancer therapy that will potential kill tumour cells more effective and selective. The use of traditional medicine as an alternative source of healthcare is gaining an increasing interest in many countries including South Africa. This is due to their accessibly and perceived effectiveness. However, the majority of these traditional medicines have not been scientifically validated. Aim: The purpose of this study was to determine immunomodulatory and anti-cancer activity of an African traditional medicine product from a traditional healer (Product Nkadinde, a traditional medicine composed of four medicinal plants) on MCF-7 breast tumour cells. Methods: Normal peripheral blood mononuclear cells (PBMCs) were used initially to establish the IC50 dose of Product Nkabinde using the Celltiter-Glo™ ATP assay. Conditioned media from PBMCs was then prepared by treating a separate set of normal PBMCs using the IC50 dose and the positive control, camptothecin (1μM). For the anticancer assays, the Celltiter-Glo™ ATP assay was used to determine the cytotoxicity effects of conditioned media from PBMCs with Product Nkabinde against MCF-7 breast cancer cells. Antioxidant levels (changes in intracellular glutathione (GSH) levels) were measured using GSH-Glo™ Glutathione assay kit. The mechanism of cell death was determined using the Caspase 3/7 and DNA fragmentation assays. Immunomodulation studies was carried out using a multi analyte cytokine ELISA assay kit. Results: The results obtained from this study revealed that Product Nkabinde was cytotoxic to MCF-7 and an IC50 dose was established at 344 μg/ml in treatment PBMCs. Product Nkabinde was able to induce caspase 3/7 activity, decrease intracellular GSH levels and induce DNA fragmentation comparative to the positive control, thus showed potential to induce apoptosis in breast cancer cells. Inflammatory cytokines secretion studies indicated that Product Nkabinde did not significantly change the secretion of all 12 inflammatory cytokines analyzed in treated PBMCs. However, Product Nkabinde was shown to increase the secretion of IL 1β, IL 8 and GM-CSF in treated MCF-7 cells, although this was not significant. Conclusion: The results of this study showed that Product Nkabinde induced immunomodulatory effects through changes in cytokines secretion and direct anticancer effects through activation of caspase 3/7 enzymes, decreasing GSH levels and induction of DNA fragmentation in treated MCF-7 cells. This traditional medicine has the potential to be developed and optimized as an alternative anticancer medicine.