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

发布时间：2018-04-09 08:00

本文选题：动脉粥样硬化　切入点：内皮细胞　出处：《中国组织工程研究》2017年01期

【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。
[Abstract]:Background: vascular endothelial cells close to the vascular wall play a vital role in the regulation of vascular homeostasis.Aim: to investigate the effects of adenovirus-mediated adiponectin overexpression on oxidative low-density lipoprotein ox-LDL- induced atherosclerotic injury of human umbilical vein endothelial cells (HUVEC) and the negative regulation of nuclear factor 魏 B p65 nuclear protein and its downstream proinflammatory factor in signal pathway.Methods: the experiment was divided into 4 groups: normal control group, normal culture of human umbilical vein endothelial cells, no ox-LDL induced by ox-LDL, 30 mg/L ox-LDL for 48 h, and dithiocarbamate pyrrolidine (PDTC) to interfere with ox-LDL induced group.The cells were treated with 100 渭 mol/L PDTC for 1 h, and 48 h after treated with ox-LDL 30 mg/L Ad-APN-eGFP. MOI=100 pfu/cell was transfected into human umbilical vein endothelial cells for 2 h, then ox-LDL 30 mg/L for 48 h.The indexes of each group were detected after intervention.Results and conclusion Human umbilical vein endothelial cells (HUVECs) were induced to injure endothelial cells and activate nuclear factor 魏 B (NF- 魏 B) signaling pathway. The expression of nuclear factor- 魏 B p65 nuclear protein was increased and the activation of NF- 魏 B pathway was inhibited by transfection of nuclear factor-kappa d-APN-eG FP.Nuclear factor 魏 B p65 decreased the expression of nuclear protein, decreased the gene and protein expression of VCAM-1, ICAM-1, and increased the gene and protein expression of endothelial nitric oxide synthase (NOS), which was regulated by nuclear factor 魏 B (NF- 魏 B).The results showed that Ad-APN-eGFP vector transfected human umbilical vein endothelial cells could effectively inhibit the activation of nuclear factor 魏 B (NF- 魏 B) in human umbilical vein endothelial cells induced by ox-LDL, and alleviate the pro-inflammatory injury and apoptosis of human umbilical vein endothelial cells induced by ox-LDL.The results showed that r Ad-APN-eGFP and PDTC could attenuate the inflammation of atherosclerosis by inhibiting the activation of nuclear factor 魏 B pathway.
【作者单位】：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;
【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)~~
【分类号】：R54

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