Intervention of Tert-butyl Hydroperoxide-Induced Injury in Human Umbilical Cord Derived Mesenchymal Stem Cells with Ginkgo Biloba Extract / 广州中医药大学学报

ABSTRACT

Objective To observe the intervention effect of Ginkgo biloba extract EGb761 on tert-butyl hydroperoxide ( t-BHP)-induced injury in human umbilical cord-derived mesenchymal stem cells ( hUC-MSCs) . Methods Proliferation of hUC-MSCs after primary culture was detected by methyl thiazolyl tetrazolium (MTT) assay, and then the optimal concentrations of t-BHP and EGb761 for oxidative stress injured MSC model were screened. Cell apoptosis was assessed by flow cytometry after Annexin V-fluorescein isothiocyanatel propidium iodide ( Annexin V-FITC/PI) staining. Content of malondialdehyde ( MDA) and superoxide dismutase (SOD) activity in hUC-MSCs were evaluated, and the expression levels of p53 and p21Cip1/Waf1 were analyzed by real-time fluorescence PCR. Results Pretreatment with 10~200 mg/L of EGb761 for 3 hours reduced the sensitivity of hUC-MSCs to t-BHP ( 100 μmol/L) induced proliferation inhibition, while EGb761 over 100 mg/L had no significant effect on enhancing the protection of hUC-MSCs . EGb761 at 100 mg/L prohibited hUC-MSCs apoptosis and MDA accumulation in hUC-MSCs induced by 100μmol/L of t-BHP acting for 6 hours, maintained the enzymatic activity of SOD, and decreased the expression of p53 and p21Cip1/Waf1 in hUC-MSCs with t-BHP-induced injury. Conclusion EGb761 is capable of protecting hUC-MSCs against oxidative stress injury, and its mechanism is probably related with the modulation of p53/p21 signal pathway.