Study on the protective effect of Tadehaginoside on the damage of endothelial cell mitochondria induced by reactive nitrogen

Objective:To investigate the protective effect of Tadehaginoside on vascular endothelial cell injury induced by reactive nitrogen.Methods:MTT colorimetry was used to detect the effect of Tadehaginoside on the survival rate of EA.hy 926 endothelial cells in the concentration range of 5~160μmol/L;1 h after pre-administration of Tadehaginoside,0.5 mM GSNO was given to damage endothelial cells.Detect the mitochondrial specific factors COX-1,ND-1 and inflammatory factor IL-1βof EA.hy 926 cells damaged by GSNO by Real time-PCR method gene intervention.At the same time,Western blot was used to detect the changes in Bax and Bcl-2 protein expression.The mitochondrial membrane potential kit(JC-1)was used to detect the change of Tadehaginoside on the mitochondrial membrane potential after GSNO induced EA.hy 926 cell injury.Results:The results of the MTT method showed that Tadehaginoside had no obvious cytotoxicity on EA.hy 926 cells in the range of 5~160μmol/L,and the optimal protective concentration of the drug was 40μmol/L.Western Blot method showed that BAX protein expression increased in a time-dependent manner after GSNO damaged EA.hy 926 cells over time,while Bcl-2 protein expression was the opposite.Real time-PCR results showed that Tadehaginoside can significantly up-regulate COX-1 gene(P<0.05),and can significantly inhibit GSNO induced ND-1(P<0.05)and IL-1βgene up-regulation(P<0.01).At the same time,the results of JC-1 showed that Tadehaginoside could significantly protect the mitochondrial membrane potential from GSNO damage.Conclusion:The GSNO damage model may induce the increase of Bax and other pro-apoptotic proteins through mitochondrial DNA damage and reduce the expression of anti-apoptotic factor Bcl-2.Tadehaginoside has a certain protective effect on endothelial cell mitochondrial damage induced by reactive nitrogen,and its mechanism is related to inhibiting the expression of ND-1 and IL-1βgenes and upregulating the expression of COX-1 genes.