Effect And Mechanism Of Transmembrane Protein 66 On Vascular Restenosis

ObjectiveCoronary atherosclerotic heart disease(CHD)is a serious hazard to human health.Percutaneous Coronary Intervention(PCI)is an important means of treating coronary heart disease.Over the past 20 years,PCI has made great progress,but the incidence of restenosis(Restenosis,RS)is still as high as 10%,reduced the clinical efficacy of PCI seriously.Therefore,to explore the mechanism of RS is to guide the prevention and control of the key.Intimal hyperplasia after vascular endothelial injury is an important pathological process of vascular restenosis.The proliferation and migration of vascular smooth muscle cells(VSMCs)are the main causes of intimal hyperplasia.Studies have shown that store operated calcium entry(SOCE)plays an important role in the proliferation,migration and intimal hyperplasia of VSMCs,Transmembrane protein 66(TMEM66)is another important component of the store-operated calcium channel(SOC),which can lead to the inactivation of SOC.Therefore,we speculate that TMEM66 may regulate VSMCs proliferation and migration after vascular endothelial injury,inhibite intimal hyperplasia,thereby reduce vascular restenosis.In order to verify the above speculation,this experiment is divided into two parts.In the first part,the model of SD and Apo E-/-rat Restenosis model of vascular injury were established by high-fat diet combined with balloon injury.The differences between the two models were analyzed.In the second part,the carotid artery restenosis model of SD rats was reproduced,and the expression of TMEM66 in blood vessels was observed.In order to verify the effect of TMEM66 on restenosis after vascular injury,we observe the changes of vascular restenosis after overexpressing TMEM66.The mechanism of TMEM66 in restenosis after vascular injury was investigated by detecting the proliferation and migration of VSMCs in different intervention groups.Methods1.The rats were divided into SD rats sham operation group,SD rats operation group,ApoE-/-rats sham operation group,ApoE-/-rats operation group.Glu,serum total cholesterol(TC),triglyceride(TG),LDL,HDL,C-reactive protein(CRP),aspartate aminotransferase(AST),alanine aminotransferase(ALT),urea nitrogen(BUN)and creatinine(Scr)were measured after 2 weeks of high-fat diet.The left common carotid artery balloon injury was performed in the two operation groups,and the balloon injury was not performed in the two sham operation groups,and the rest of the operation was the same as the operation group.H&E staining observed plaque size,oil red O staining,MASSON staining,immunofluorescence staining detected the plaque composition.2.SD rats were randomly divided into control group(sham operation),operation group(left common carotid artery balloon injury),No-load virus group(No-load virus transfection after left common carotid artery balloon injury),TMEM66 overexpression group(TMEM66 gene overexpression transfection after left common carotid artery balloon injury).The Real-time fluorescence quantitative PCR and Western blot were used to detect the expression of TMEM66 in sham operation group and operation group.The expression of TMEM66 was observed by immunohistochemical staining.H&E staining observed each group of vascular restenosis.3.VSMCs were randomly divided into control group,PDGF treatment group(PDGF treatment),No-load virus group(PDGF treatment after no-load virus transfection),TMEM66 overexpression group(PDGF treatment after TMEM66 gene overexpression),EdU staining was used to detect VSMCs proliferation ability,scratch test was used to detect VSMCs migration ability.Results1.Blood cholesterol and inflammatory mediators of ApoE-/-Rats were significantly higher than those in SD rats.Two groups of rats had significant restenosis plaque formation.Lipid infiltration and fibrous deposition were detected in restenosis plaque of Apo E-/-rats,while cell components were mainly of macrophages and VSMCs.However,VSMCs were the main components of vascular restenosis in SD rats.2.The expression of TMEM66 was significantly decreased after vascular endothelial stenosis,and the overexpression of TMEM66 could inhibit the restenosis after endothelial injury.3.Upregulation of TMEM66 expression can significantly inhibit the proliferation and migration of VSMCs.Conclusion:Transmembrane protein 66 may inhibit hyperplasia of neointima,thereby reduce vascular restenosis after endothelial injury via inhibiting the proliferation and migration of VSMCs.