重组腺病毒介导脂联素对人脐静脉内皮细胞损伤的作用和机制  

Effects of recombinant adenovirus-mediated adiponectin on human umbilical vein endothelial cell injury and the underlying mechanism

作　　者：王雪梅[1] 魏琴[1] 段明军[1] 张春[1] 杨毅宁[1] 

机构地区：[1]新疆医学动物模型研究重点实验室,新疆医科大学第一附属医院临床医学研究院,新疆维吾尔自治区乌鲁木齐市830011

出　　处：《中国组织工程研究》2017年第1期115-121,共7页Chinese Journal of Tissue Engineering Research

基　　金：新疆维吾尔自治区自然科学基金青年基金项目(2015211C095);新疆重大疾病医学重点实验室-省部共建国家重点实验室培育基地开放课题(SKLIB-XJMDR-2014-16)~~

摘　　要：背景:血管内皮细胞紧贴于血管壁,在血管稳态平衡的调节中发挥着至关重要的作用。目的:分析腺病毒介导的脂联素过表达对氧化型低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞动脉粥样化损伤、核因子κB p65核蛋白及其信号通路下游促炎因子的负性调控作用。方法:实验分4组:正常对照组,采用人脐静脉内皮细胞常规培养,未用ox-LDL诱导;ox-LDL诱导组,用30 mg/L的ox-LDL诱导人脐静脉内皮细胞48 h;二硫代氨基甲酸吡咯烷(PDTC)干预+ox-LDL诱导组,先用100μmol/L的PDTC处理细胞1 h,再用ox-LDL 30 mg/L诱导48 h;Ad-APN-eGFP干预+ox-LDL诱导组,以MOI=100 pfu/cell转染人脐静脉内皮细胞2 h后,用ox-LDL 30 mg/L诱导48 h。各组干预后检测各项指标。结果与结论:(1)ox-LDL诱导人脐静脉内皮细胞造成内皮细胞损伤模型,激活核因子κB信号通路,核因子κB p65核蛋白的表达量增加;(2)rA d-APN-eG FP转染能够抑制核因子κB通路的激活,使核因子κB p65核蛋白表达量降低,降低核因子κB调节的下游促炎分子VCAM-1、ICAM-1、MCP-1的基因和蛋白表达,增加抗炎因子内皮型一氧化氮合酶的基因和蛋白表达,差异有显著性表达;(3)结果说明,r Ad-APN-eGFP载体转染人脐静脉内皮细胞,通过有效抑制ox-LDL诱导人脐静脉内皮细胞的核因子κB激活,减轻ox-LDL诱导的细胞促炎损伤和细胞凋亡,与使用PDTC干预具有相近的效果;(4)结果证实r Ad-APN-eGFP和PDTC一样可通过抑制核因子κB通路的激活来减轻动脉粥样硬化这种炎症反应。BACKGROUND： Vascular endothelial cells closely adhere to the blood vessel wall so as to play a crucial role in the regulation of vascular homeostasis, OBJECTIVE： To analyze the inhibitory effects of recombinant adenovirus-mediated adiponectin overexpression on oxidized low density lipoprotein （ox-LDL）-induced atherosclerotic injury in human umbilical vein endothelial cells, nuclear factor KB （NF-KB） p65 nuclear protein and pro-inflammatory factors in the downstream of signaling pathway. METHODS： There were four groups in this experiment： human umbilical vein endothelial cells were cultured in the conventional medium without ox-LDL induction as control group; human umbilical vein endothelial cells induced by 30 mg/L ox-LDL for 48 hours as ox-LDL induction group; cells firstly treated with 100 pmol/L pyrrolidine dithiocarbamate （PDTC） for I hour, and then induced by 30 mg/L ox-LDL for 48 hours as PDTC with ox-LDL induction group; cells transfected by adenovirus bearing a vector encoding for adiponectin and enhanced green fluorescent protein （Ad-APN-eGFP） （multiplicity of infection=100 pfu/cell） for 2 hours, and then induced by 30 mg/L ox-LDL for 48 hours as Ad-APN-eGFP With ox-LDL inductiongroup. Afterwards, all relative indicators were detected in each group. RESULTS AND CONCLUSION： Endothelial cell injury models were obtained by ox-LDL induction, and the NF-KB signaling pathway was activated with the increased expression of NF-KB p65 nuclear protein. Ad-APN-eGFP could inhibit the NF-KB signaling pathway, which significantly decreased the expression of NF-KB p65 nuclear protein, as well as the gene and protein expressions of VCAM-1, ICAM-1 and MCP-1 in the downstream of signaling pathway, but significantly increased the gene and protein expressions of endothelial nitric oxide synthase. These results show that Ad-APN-eGFP-tranfected human umbilical vein endothelial cells can relieve ox-LDL-induced pro-inflammatory injury and reduced apoptosis via effectively inhibiting the NF-KB activa

关 键 词：组织构建 内皮细胞 脂联素 动脉粥样硬化 人脐静脉内皮细胞 氧化型低密度脂蛋白 NF-κB信号通路 新疆维吾尔自治区自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]