Investigation of ursodeoxycholic acid effects on sirolimus treated and adipose derived mesencyhmal stem cell line

Mesenchymal stem cell therapies are important therapeutic modalities that have been applied experimentally and clinically in many fields of medicine today. Transplantation of mesenchymal stem cells into patients for supportive purposes besides immunosuppressive drugs is becoming common after organ transplantation in clinical cases. The aim of this study is to investigate the cytotoxic effects of Sirolimus, which is used for immunosuppressive purposes in organ transplants, on mesenchymal stem cells derivating from human adipose tissue and to investigate the effects of ursedeoxycholic acid (UDCA), which is known to have antioxidant properties, on this cytotoxicity. The human adipose tissue-derived mesenchymal stem cell line is commercially available. Human mesenchymal stem cells produced in special media which were incubated at 37 ° C until sufficient numbers were reached. The cytotoxic effects of the active substances were assessed by time and dose-dependent WST-1 test. Light microscopy and cell counting device were used for vitality and proliferation analyzes. Muse was used for evaluation of apoptosis and cell cycle in cell analyzer. Lipid peroxidation, superoxide dismutase (SOD) activity, catalase (CAT) activity, glutathione peroxidase (GPX) activity were measured by biochemical methods. Cytotoxic doses of the agents administered to the groups were found by the WST test. Non-apoptotic, early apoptotic, late apoptotic, total apoptotic and dead cells were calculated as both percent and cell/ml by Annexin V. Lipid peroxidation, SOD activity, CAT activity, GPX activity were measured to determine the damage caused by oxidative stress and reactive oxygen metabolites. In human adipose tissue-derived mesenchymal stem cells, Sirolimus IC50 was determined as 15.85 μM for dozen (48th hour). For UDCA, no cytotoxic effect was observed at the doses studied. Therefore, the study continued with the highest fixed dose of 100 μM. In the control group, according to the measurements made with Muse® cell analyzer the calculations made, the dead cell was calculated as 1.03% and the living cell as 98.97%. In the UDCA group, dead cells were calculated as 2.59% and living cells as 97.41%. In the Sirolimus group, early apoptosis was calculated as 7.69%, late apoptosis as 4.31%, dead cell as 8.10% and live cell as 79.90%. In the combined group of Sirolimus+ UDCA early apoptosis rate was 3.90%, late apoptosis rate was 1.34%, dead cell was 7.81% and viable cell was 86.95%. In the cell cycle analysis, the percentages of the G2/ M phases in the control and UDCA groups were 19.3% and 18.9%, respectively. The percentage of G2/ M phase in the Sirolimus treatment group was reduced to 15.9. Biochemical findings were; MDA- combined group; All binary groups except for Control- UDCA in terms of GSH-Px levels; Control in terms of CAT levels- UDCA; control-combined; UDCA- Sirolimus and UDCA- combined groups; SOD levels were statistically significant (p <0.001). In the study, Sirolimus was found to be compatible with the literature in promoting apoptosis in mesencymal stem cells. The presence of evidence that UDCA regulates cell proliferation and the antiapoptotic effects of UDCA have been shown to be consistent with suppressing the Sirolimus effect in the combined group. In immunosuppressive treatment after organ and tissue transplantation and in adipose tissue-derived mesenchymal stem cell application, the use of UDCA showed positive effects on mesenchymal stem cells.