Effect Of MiR-630 Regulating JAK/STAT Signaling Pathway On Proliferation/Apoptosis By Targeted IRS1 In HUVECs

Objective:People with atherosclerosis(AS)have a significantly increased risk of cardiovascular disease in the future.The atherosclerotic process begins in childhood and progresses silently after a long period of preclinical stage,usually beginning in middle age and eventually manifesting clinically.In many cases,when the disease has fully developed,the clinical manifestations of atherosclerosis will appear,which will lead to significant narrowing of the vascular lumen and possibly fatal consequences.Therefore,finding new mechanisms to better understand the pathogenesis of atherosclerosis can provide new diagnostic biomarkers and therapeutic targets for the AS process.AS is a complex systemic inflammatory vascular disease.Vascular endothelial dysfunction can lead to the dynamic development of atherosclerotic plaques.Therefore,it is necessary to further determine the important regulatory molecules involved in endothelial function and their regulatory pathways in future therapeutic applications.The differential expression of miRNA in AS is related to the proliferation,apoptosis,inflammation,angiogenesis and remodeling of endothelial cells.Although many miRNAs have been reported to participate in the process of regulating AS,the mechanism of miRNA’s influence on endothelial cell function is not completely clear,and further research is needed.Method:This project uses the Gene Expression Omnibus(GEO)database for data mining to analyze the differential expression of miRNA and mRNA in AS.Endothelial cell proliferation,apoptosis,and angiogenesis were analyzed by CCK8,flow cytometry,and in vitro angiogenesis;the combination of miRNA and mRNA was detected by luciferase experiment;the expression of miRNA was detected by RT-qPCR,and the protein was detected by western blot detection.Result:GSE137580 samples found that there were 26 differentially expressed miRNAs in human aortic endothelial cells treated with oxidized low-density lipoprotein,of which 19 were up-regulated and 7 were down-regulated.The expression of miR-630 was up-regulated in human aortic endothelial cells treated with oxidized low-density lipoprotein.Overexpression of miR-630 can promote the apoptosis of HUVECs and inhibit their proliferation and angiogenesis;whereas interference with the expression of miR-630 can inhibit the apoptosis of HUVECs cells promotes their proliferation and angiogenesis.Targetscan analysis and luciferase experiment showed that miR-630 can bind to the 3’-untranslated region of IRS1.Overexpression of miR-630 can inhibit the expression of IRS1 while interference with miR-630 promotes the expression of IRS1.Interference with the expression of IRS1 can promote the apoptosis of HUVECs,inhibit their proliferation and angiogenesis,and reverse the protective effect of miR-630 silencing on HUVECs.Overexpression of miR-630 can activate JAK / STAT signaling while interference with miR-630 inhibits JAK / STAT signaling.Additionally,interference with IRS1 can activate JAK / STAT signaling,reversing the protective effect of miR-630 silencing on HUVECs.Conclusion:miR-630 activates the JAK / STAT signaling pathway by targeting the expression of IRS1 to promote the apoptosis of HUVECs,inhibit their proliferation and angiogenesis,thereby promoting the occurrence of AS.The elucidation of this mechanism provides a potential target for the treatment of AS.