Investigation of The Effect of Oxytosin on Mesenchymal Stem Cells Exposed to Sunitinib
Project no. 21772 · Status: Kapanmış · 2020
Summary
Mesenchymal stem cells are essential treatment methods used in experimental and clinical studies. Nowadays, mesenchymal stem cell therapy is used in many various diseases. In addition to immunosuppressive drug therapy used after organ transplants, a mesenchymal stem cell application can also be utilized in the patient's treatment. In this experimental study, the cytotoxic effects of Sunitinib drug, which is used for immunosuppressive purposes in organ transplants, on mesenchymal stem cells which is derivated from adipose tissue and the toxicity effect on oxytocin (known to have antioxidant properties) were inspected. A mesenchymal stem cell line derived from mouse adipose tissue was commercially available. In line with the work to be done, stromal stem cells were incubated at 37°C in a special medium until they reached a sufficient number. The cytotoxic effects of the active ingredients were evaluated using the WST-1 assay in a time and dose-dependent manner. Viability and prevalence analyze were determined using light microscopy and cell counting. Apoptosis and cell viability were assessed with a BD Accuri C6 flow cytometer analyzer. Lipid peroxidation, glutathione peroxidase (GPx), catalase activity (CAT), and superoxide dismutase (SOD) activity were measured by biochemical methods. The cytotoxic doses of agents applied to the groups were assessed by the WST-1 assay. Necrotic, early apoptotic, late apoptotic, total, and viable cells were calculated in both percentage and cell/ml by the Annexin V method. The effect of lipid peroxidation, glutathione peroxidase (GPx), catalase activity (CAT), and superoxide dismutase activity (SOD) was measured in order to determine the damage due to oxidative stress and reactive oxygen metabolites. The IC50 dose of Sunitinib in mesenchymal stem cells derived from adipose tissue of mice (48 h) was calculated as 44.57 µM. For oxytocin, no cytotoxic effect was observed at the studied doses. Therefore, the study continued with the highest stable dose of 100 µM. According to the measurements and calculations made with the BD Accuri C6 cellular device, the percentage of dead cells in cells in the control group was calculated at 1.03 and the percentage of live cells was 98.97. The percentage of dead cells in the cells in the oxytocin group was 2.59 and the percentage of live cells was 97.41. In the sunitinib group, the rate of early apoptosis was 7.69%, late apoptosis rate 4.31%, dead cells 8.10%, and live-cell 79.90%. In the results of the pooled Sunitinib + oxytocin group, the rate of early apoptosis was 3.90%, the rate of late apoptosis was 1.34%, the percentage of dead cells 7.81, and the percentage of live cells 25.5. As for the biochemical results, confidence intervals were calculated according to the other groups as there was no change in MDA levels in the control group, GSH-Px levels in the Sunitinib group, and CAT levels in the oxytocin group, and there was no significant difference between the groups. All marital differences between all groups were statistically significant (p<0.001). In this study, as a result of a review of the literature, Sunitinib is expected to increase apoptosis over mesenchymal stem cells. There are results showing that oxytocin regulates cell proliferation, but the anti-inflammatory effects of oxytocin were found unable to suppress the sunitinib effect of oxytocin in the co-group for this study.