Study On The Protective Mechanism Of Tanshinone ⅡA On Endothelial Cells Caused By Hypoxia

Purpose:1 To study the mechanism of tanshinone IIA on human aortic endothelial cells deprived of oxygen and sugar,and to explore whether tanshinone IIA can increase e NOS expression in endothelial cells,induce NO synthesis and protect the function of damaged endothelial cells.2 To explore whether tanshinone IIA can improve the function of damaged vascular endothelial cells by regulating the expression of micro RNA-24 and activating the expression of e NOS in vascular endothelial cells.Material and method:1 Human aortic endothelial cells(HAECs)were purchased by biological companies,and then subcultured in dimethylformamide(DMEM)medium containing 10% fetal bovine serum FBS with fusion rate of 70-80%.2 HAECs cultured with 10% FBS and high-sugar DMEM at logarithmic growth stage were used to establish a hypoglycemic anaerobic model.3 HAECs were cultured in DMEM medium with different concentrations of TSA(0,10,30,50 mg/ml)for 24 hours to establish TSA intervention group.4 HAECs were divided into blank control group,hypoglycemic anaerobic group and TSA administration group.5 Cell morphology was observed by electron microscopy after 48 hours of culture in different ways in each group.The cell morphology of TSA group after 48 hours was significantly better than that of anaerobic group after 48 hours of glucose deficiency.6 MTT assay was used to detect the survival rate of endothelial cells.7 Cell supernatant was collected,and the content of protein e NOS in cell supernatant was detected by ELISA.The expression of e NOS was detected by Western blot and q RT-PCR.8 Quantitative RT-PCR was used to detect the expression of micro RNA-24 in cells.Results:1 Cell morphology of TSA treated groups was significantly improved compared with that of glucose-deficient and anaerobic groups.2 The survival rate of HAECs in TSA groups was significantly higher than that in glucose-deficient and anaerobic groups.3 The expression of e NOS in HAECs increased significantly in all groups of TSA administration.4 The expression of micro RNA-24 gene in HAECs of TSA groups was significantly down-regulated.Conclusion:1 TSA can improve the morphology of vascular endothelial cells.2 TSA can improve the survival rate of damaged endothelial cells in blood vessels.3 TSA can increase the expression of protein e NOS in damaged vascular endothelial cells.4 TSA can down-regulate the expression of micro RNA-24 in vascular endothelial cells.