Prdx2 Of Expression In Human Carotid Atherosclerosis And Its Effect On The Proliferation And Migration Of Vascular Smooth Muscle Cells

Background: Atherosclerosis(AS)is a chronic disease of the arterial wall.Atherosclerotic plaques and other complex lesions are formed within the artery due to lesions,mainly involving large and medium-sized arteries.The basic lesions of AS are arterial intima lipid deposition,endometrial focal fibrosis,atheromatous plaque formation,hardening of the wall,and narrowing of the lumen,causing ischemic changes in the corresponding organ.Inflammation-injury-response theory has been widely supported and accepted.It has been confirmed in literature that endothelial cell injury secretes growth factors,which activates arterial smooth muscle cells,migrates into the tunica intima via the inner elastic membrane,and undergoes proliferation,transforms,secretes cytokines and synthesizes extracellular matrix.Finally,smooth muscle cells engulf lipids to form smooth muscle cell-derived foam cells.Both tumor necrosis factor-?(TNF-?)and angiotensin II(AngII)have chemical chemotaxis,which increase intracellular ROS content,thereby inducing proliferation and migration of vascular smooth muscle cells and accelerating the occurrence and development of AS.In addition,proliferation of cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP2)and matrix metalloproteinase-9(MMP9)are involved in the biological behavior of vascular smooth muscle cells.Prdx2 has an important function of scavenging reactive oxygen species(ROS),while ROS participates in the proliferation,migration and apoptosis of vascular smooth muscle cells.Reactive oxygen species have been shown to accelerate the development of AS,but little is known about the role and mechanism of Prdx2 in atherosclerosis and remains to be further studied.Mitogen-activated kinase(MAPK)is an important regulatory enzyme that links the receptors on the surface of cell membrane with a definitive gene expression and is one of the important signal transduction pathways in the cell.MAPK signaling pathway plays an important regulatory role in the pathological process of various cardiovascular diseases.Methods:1.Immunohistochemistry was used to detect the differential expression and localization of Prdx2 in human carotid atherosclerotic tissues.2.The qPCR method and Western-blot method were used to detect the mRNA and protein expression of Prdx2 in human carotid normal tissue and human carotid atherosclerosis.3.Reactive oxygen species assay kit,CCK8 assay and Western-blot assay detect intracellular ROS content and cell proliferation after stimulation of human carotid vascular smooth muscle cells with tumor necrosis factor alpha(TNF-?)and angiotensin II(Ang II)Conditions and Prdx2 protein expression levels.4.Reactive oxygen species assay kit,CCK8 assay and Western-blot assay detected ROS content,cell proliferation,and Prdx2 protein expression in human carotid vascular smooth muscle cells stimulated by TNF-?+NAC and AngII+NAC.5.qPCR method,Western-blot method and cell immunofluorescence method were used to detect the interference efficiency of Prdx2 siRNA against human carotid artery smooth muscle cells.6.The changes of proliferation and migration of human carotid vascular smooth muscle cells after Prdx2 gene interference were detected by CCK8 assay,cell scratch assay and Transwell assay.7.Western-blot was used to detect the protein expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP2)and matrix metalloproteinase-9(MMP9)in human carotid vascular smooth muscle cells after Prdx2 gene interference.8.Using reactive oxygen detection kit to detect the changes of ROS content in human carotid vascular smooth muscle cells after Prdx2 gene interference.9.CCK8 assay and Transwell cell migration assay were used to detect the changes of the proliferation and migration of human carotid artery smooth muscle cells after antioxidation of NAC on Prdx2.10.The expression of MAPKs pathway-related factors JNK1/2,p38 and ERK1/2 phosphorylation after Prdx2 gene interference with human carotid vascular smooth muscle cells was detected by Western-blot.Results:1.Prdx2 was highly expressed in the human carotid atherosclerotic tissues compared with normal human carotid tissues at the protein level and mRNA level(P<0.05).Prdx2 was highly expressed in smooth muscle cell-derived cell regions of plaques.2.TNF-? and AngII stimulated carotid vascular smooth muscle cells to increase ROS content,increase cell proliferation and increase Prdx2 expression significantly(P<0.05).3.After the antioxidant NAC acted on the atherosclerosis model in vitro,the ROS content of human carotid artery smooth muscle cells was significantly decreased,the cell proliferation ability was decreased,and the Prdx2 expression was significantly downregulated(P<0.05).4.Compared with the negative control group,the proliferation and migration of human carotid vascular smooth muscle cells were significantly increased after Prdx2 gene interference(P<0.05).5.Compared with the negative control group,the expression of PCNA,MMP2 and MMP9 increased after Prdx2 gene interfered with human carotid vascular smooth muscle cells(P<0.05).6.Reactive oxygen species detection showed that Prdx2 gene interfered with human carotid vascular smooth muscle cells and the content of endogenous ROS increased significantly(P<0.05).7.Antioxidant NAC acted on human carotid vascular smooth muscle cells silenced by Prdx2 gene for 24 h,the cell proliferation and migration ability was significantly decreased(p<0.05).8.Antioxidant Western-blot results showed that the expression of p-JNK1/2,p-p38,and p-ERK1/2 protein was significantly increased in Pdhx2 siRNA-transfected human carotid vascular smooth muscle cells 48 hours after treatment with Prdx2 siRNA.(P<0.05).There was no correlation between the total protein expression of JNK1/2 and p38.Conclusion: Prdx2 gene has a significant increase in the proliferation and migration of human carotid vascular smooth muscle cells,which may be related to the increase of intracellular ROS content after Prdx2 silencing.Prdx2 is expressed in the smooth muscle cell-derived cells of human carotid atherosclerotic tissue.High expression,suggesting that Prdx2 high expression may not be pathogenic,may be due to the body's negative feedback regulation.