| Objective:Type A Aortic Dissection（TAAD）is a kind of vital dangerous cardiovascular disease.Stanford type A aortic dissection extend over the ascending aorta,it may also involve the aortic arch,descending aorta and abdominal aorta.Death rates of TAAD are extremely high[1].There’s no clear cause of TAAD,a possible inducement is the of aortic wall atherosclerosis[2].Atherosclerosis is a chronic inflammatory vascular disease.It can involve vascular endothelial injury,cell proliferation and migration of smooth muscle,lipid accumulation in intima and plaque formation.Sirtuin 1（SIRT1）is currently recognized as a longevity gene,also one of the most studied members of the Sirtuin family.SIRT1 participate in the regulation of glycolipid metabolism,inflammation,oxidative stress,atherosclerosis and other physiological and pathological process[3].Of course,SIRT1expression is also regulated by a variety of m RNA.MiR-487a-3p and SIRT1 have possible regulatory relationships predicted by bioinformatics.Studies have shown that miR-487a-3p highly expressed in a variety of malignancies[6],but less research in cardiovascular disease.This paper aims to study whether miR-487a-3p can play a role in atherosclerosis-induced TAAD by regulating SIRT1.Methods:1.Collecting the aortic wall tissue of patients with aortic dissection and coronary heart disease treated by surgery.TAAD was divided into atherosclerosis group（AS group）and simple aortic dissection group（AD group）,the coronary heart disease group was regarded as control group（NC group）.2.The expression of miR-487a-3p in AS group,AD group and NC group.3.Western Blot was used to detect the SIRT1 expression in the above groups.4.Culturing of Human Aortic Smooth Muscle Cells（Hu VSMCs）.Oxidized low density lipoprotein（OX-LDL）was used to stimulate Hu VSMCs to acquire the model of cell atherosclerotic.CCK-8 kit was used to detect the optimal concentration and time of cell atherosclerotic.Oil red O staining was applied to examine lipid deposition in Hu VSMCs.RT-q PCR was used to detect the expression of miR-487a-3p in cells at different time.5.Dual-luciferase assay was used to verify the targeting relationship between miR-487a-3p and SIRT1.6.After transfection of miR-487a-3p mimics and miR-487a-3p mimics NC,miR-487a-3p inhibitor and miR-487a-3p inhibitor NC,we detected transfection efficiency and expression of SIRT1 m RNA by RT-q PCR.The SIRT1,MCP-1 and MMP2 expression in each transfection group and normal group were detected by Western Blot.7.Western Blot was used to detect the expression of SIRT1,MCP-1 and MMP2 after the OX-LDL after transfecting miR-487a-3p.8.The expression level of SIRT1m RNA was detected by RT-q PCR after transfection and adding OX-LDL,and the expression of SIRT1,MMP-2,MCP-1 in each group was tested by Western Blot.Results:1.Compared to NC group,MiR-487a-3p was highly expressed in AS group and in AD group MiR-487a-3p was also high.2.The result of CCK-8 result showed the best drug concentration was50μg/ml and the best time is 24h.3.In Oil red O staining,Lipid deposition was most significant in Hu VSMCs treated with 50μg/ml OX-LDL after 24h.4.RT-q PCR was used to detect the peak time of miR-487a-3p expression after 50μg/ml OX-LDL in cells and the time was 24 h.5.The double luciferase report showed that there was a negative relationship between miR-487a-3p and SIRT1.6.overexpression miR-487a-3p can reduce the SIRT1 m RNA expression level and SIRT1 protein expression level,improve the MCP-1,MMP-2protein level.The down regulated miR-487a-3p had the opposite result.Conclusion:1.The expression of miR-487a-3p is higher in the aortic dissection with atherosclerosis.2.There is negative relationship between miR-487a-3p and SIRT13.overexpression/inhibition miR-487a-3p can affect the process of atherosclerosis by regulating SIRT1. |
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