Study On The Mechanism Of MicroRNA-133a Mediate Endothelial Cell Apoptosis And Intervention Of Salidroside

BackgroundVascular endothelial cell apoptosis promotes the formation of atherosclerosis(AS),which causes a series of cardiovascular diseases and becomes a great threat to human health.microRNA-133a(miR-133a)is a kind of non-coding RNA,that participates in the pathological process of AS,but the specific molecular mechanism is unknown Salidroside(SAL),a traditional Chinese medicine,has a significant anti-AS effect,but the pharmacological mechanism is still unknownObjective1.To explore the role of miR-133a in endothelial cell apoptosis,which will provide useful data for the study of the pathological mechanism of non-coding RNA regulating endothelial injury2.To study salidroside's inhibition of endothelial cell apoptosis and explore its molecular mechanism,and provide a new approach and target for traditional Chinese medicine to prevent and treat atherosclerosisMethod1.Human coronary endothelial cells(HCAECs)were cultured and treated with oxidized low density lipoprotein(ox-LDL),cell viability was detected by MTT method,miRNA(miR-133a,etc.)was detected by qRT-PCR,and Bcl-xl mRNA levels,Western Blot detection of anti-apoptotic protein Bcl-xl and activated cysteine caspase 3(cleaved-caspase 3)expression levels,flow cytometry to detect apoptosis rate.Transient transfection technology was used to overexpress and silence miR-133a,and observe its effect on the expression of target gene Bcl-xl protein and endothelial cell apoptosis2.HCAECs were cultured and treated with SAL,cell viability was measured by MTT method,and lactate dehydrogenase(LDH)release test was used to detect cytotoxicity.In the ox-LDL-induced HCAECs apoptosis model,SAL was added for intervention,qRT-PCR was used to detect the mRNA levels of miR-133a and Bcl-xl,Western Blot was used to detect the expression of anti-apoptotic proteins Bcl-xl and cleaved-caspase 3,Flow cytometry was used to detect the apoptotic rate.Transient transfection technology overexpression and silencing miR-133a is divided into:mimics+ox-LDL group,mimics+ox-LDL+SAL group and inhibitor+ox-LDL group,inhibitor+ox-LDL+SAL group,and further detection of apoptosis and Bcl-xl protein expressionResult1.ox-LDL can reduce the viability of HCAECs cells and induce apoptosis in HCAECs;miR-133a is abnormally increased in apoptosis models;overexpression of miR-133a,Bcl-xl is reduced,and the rate of apoptosis is increased;The decrease of Bcl-xl induced by LDL and the increase of apoptosis rate were partially reversed,and the difference was statistically significant2.ox-LDL induces an increase in miR-133a,a decrease in the anti-apoptotic protein Bcl-xl,and the activation of caspase 3 in HCAECs,which can be reversed after SAL intervention;the anti-apoptotic effect of SAL is weakened after miR-133a is silenced,and overexpression of miR-133a The anti-apoptotic effect of SAL was partially recovered,and the difference was statistically significantConclusion1.miR-133a regulates apoptosis of endothelial cells induced by anti-apoptotic protein Bcl-xl2.SAL can inhibit the increase of miR-133a induced by ox-LDL,promote the expression of anti-apoptotic protein Bcl-xl,prevent endothelial cell apoptosis,protect vascular function,and resist AS.