Adipose Differentiation-related Protein Knockdown Inhibits Vascular Smooth Muscle Cell Proliferation And Migration And Attenuates Neointima Formation

Background:The morbidity of cardiovascular,cerebrovascular andperipheral vascular diseaseincreased year by year,and the mortality ranked first in the nation.These are current researching focuses that the pathogenic mechanism and the solutions of atherosclerosis.At present,with the progress of interventional techniques and the inventions of new equipment,endovascular therapy for arteriosclerosis occlusive disease has been used more and more.But,the main problem is that artery restenosis postoperative has been unable to completely solved.Vascular smooth muscle cells?VSMCs?have an important role in atherosclerosis development.VSMCs also have an important role in restenosis postoperative.Evidence has demonstratedthat adipose differentiation-related protein?ADRP?is associated with foam cell formation and atherosclerosis progression.However,to the best of our knowledge,no previous studies have investigated the role of ADRP knockdown in platelet-derived growth factor?PDGF?-stimulated proliferation and migration of VSMCs in vitro.Furthermore,the effect of ADRP knockdown on neointima formation in vivo remains unclear.Objective:We investigate the effects of ADRP knockdown on arteriosclerosis model mice in vivo.We investigate the effects of ADRP knockdown on neointima formation after vascular injury by balloon in vivo.Weinitiatethecurrentstudyto determinethe role of ADRP knockdown in platelet-derived growth factor?PDGF?-stimulated proliferation and migration of VSMCs in vitro.Weexplore themolecular mechanism of ADRP how to influence on endometrial hyperplasia and VSMC.Methods:1.VSMCs were incubated with PDGF following ADRP small interfering?si?RNA transfection.Cell viability,migration and cell cycle distribution were analyzed by MTT and Transwell assays and flow cytometry,respectively.Extracellular signal-regulated kinase?ERK?,phosphorylated?p?-ERK,Akt,p-Akt,proliferating cell nuclear antigen?PCNA?,matrix metalloproteinase?MMP?-2 and MMP-9 protein levels were determined by western blotting.2.Apolipoprotein E^-/-mice fed an atherogenic diet were injected with siADRP or control siRNA.After 3 weeks,aortas were excised and mRNA and protein ADRP expression was determined by RT-qPCR and western blotting.Neointima formation was assessed by hematoxylin and eosinstaining.3.The restenosis model of rat's carotid artery was established by balloon injury.The rats were injected with siADRP or control siRNA.After 3 weeks,the carotid artery were excised and mRNA and protein ADRP expression was determined by RT-qPCR and western blotting.Neointima formation was assessed by hematoxylin and eosinstaining.Results:1.ADRP knockdown significantly inhibited PDGF-induced increases in VSMC viability,caused G1 phase cell cycle arrest and decreased PCNA expression.Knockdown of ADRP inhibited PDGF-induced migration of VSMCs by reducing MMP protein expression and activity.In addition,the present study also demonstrated that ADRP knockdown inhibited ERK and Akt signaling pathways in response to PDGF.Furthermore,si ADRP administration suppressed neointima formation in model mice.2.The results of the current study demonstrated that the expression of ADRP mRNAand ADRP were increased in the arteriosclerosis model mice.Subintimal lipid deposited,plaque formated,VSMCs proliferated and migrated to the endometrium,these were observed under hematoxylin and eosinstaining.siADRP administration suppressed the expression of m RNA and protein ADRP in model mice.siADRP administration suppressed neointima formation in model mice,too.3.The results of the current study demonstrated that the expression of ADRP mRNA and ADRP were increased in the restenosis model rats.VSMCs proliferated and migrated to the endometrium,neointima formation of model rats,these were observed under hematoxylin and eosinstaining.siADRP administration suppressed the expression of mRNA and protein ADRP in model rats.siADRP administration suppressed neointima formation in model rats,too.Conclusions:ADRP knockdown significantly inhibited PDGF-induced increases in VSMC viability,caused G1 phase cell cycle arrest,inhibited ERK and Akt signaling pathways in response to PDGF.ADRP knockdown administration suppressed neointima formation in arteriosclerosis model mice.ADRP knockdown administration suppressed neointima formation inrestenosis model rats.The results of the present study indicate that ADRP may be a potential target for the treatment of atherosclerosis.