Irisin/PPARα信号通路对小鼠主动脉平滑肌细胞增殖的作用机制  

Irisin/peroxisome proliferator-activated receptor alpha signaling pathway mediates vascular smooth muscle cell proliferation in mice

作　　者：王尧 刘大男 赵广建 周博 王向 Wang Yao;Liu Danan;Zhao Guangjian;Zhou Bo;Wang Xiang(Department of Cardiology,Affiliated Hospital of Guizhou Medical University,Institute of Medical Science,Guizhou Medical University,Guiyang 550004,Guizhou Province,China)

机构地区：[1]贵州医科大学附属医院心内科,贵州医科大学医学科学研究所,贵州省贵阳市550004

出　　处：《中国组织工程研究》2023年第33期5320-5326,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81660083),项目负责人:刘大男;贵州省科技创新人才团队项目[黔科合平台人才(2020)5014],项目负责人:刘大男;贵州省“百”层次创新型人才培养计划项目[黔科合人才(2015)4026号],项目负责人:刘大男;贵州医科大学国家自然科学基金培育项目(TJ20073),项目负责人:刘大男。

摘　　要：背景:鸢尾素(Irisin)由Ⅲ型纤连蛋白结构域包含蛋白5(fibronectin typeⅢdomain containing 5,FNDC5)经水解修饰后释放,除调节糖脂代谢的主要生理功能外,同时具有调节多种细胞增殖与凋亡的能力。目的:探讨Irisin/过氧化物酶体增殖物激活受体α(peroxisome proliferator-activated receptor alpha,PPARα)信号通路介导血红素氧合酶1调节血管平滑肌细胞增殖与凋亡平衡的作用及机制。方法:采用FNDC5过表达和沉默慢病毒载体转染血管平滑肌细胞增加或抑制Irisin生成,构建体外细胞模型,分为7组:空白对照组、过表达空载体组、FNDC5组、过表达空载体+GW6471(PPARα抑制剂)组、FNDC5+GW6471组、沉默空载体组及sh-FNDC5组,采用实时荧光定量PCR和Western blot分别检测各组FNDC5、PPARα、血红素氧合酶1、Bax、Bcl-2 mRNA和蛋白表达水平;采用流式细胞技术检测血管平滑肌细胞凋亡情况,CCK-8法检测细胞增殖活性。结果与结论:(1)与空白对照组、过表达空载体组比较,FNDC5组、FNDC5+GW6471组FNDC5、PPARα、血红素氧合酶1、Bax表达升高,Bcl-2表达降低,Bax/Bcl-2比值升高(均P<0.05);(2)与空白对照组、沉默空载体组比较,sh-FNDC5组FNDC5、PPARα、血红素氧合酶1、Bax表达降低,Bcl-2表达升高,Bax/Bcl-2比值降低(均P<0.05);(3)与FNDC5组比较,FNDC5+GW6471组PPARα、血红素氧合酶1、Bax显著降低,Bcl-2表达升高,Bax/Bcl-2比值降低(均P<0.05);(4)与空白对照组、过表达空载体组比较,FNDC5组、FNDC5+GW6471组血管平滑肌细胞增殖活性显著降低,凋亡率显著升高(均P<0.05);(5)与空白对照组、沉默空载体组比较,sh-FNDC5组血管平滑肌细胞增殖活性升高,凋亡率显著降低(均P<0.05);(6)与FNDC5组比较,FNDC5+GW6471组血管平滑肌细胞增殖活性显著升高,凋亡率显著降低(均P<0.05);(7)提示Irisin/PPARα信号通路可通过诱导血红素氧合酶1表达,抑制氧化低密度脂蛋白诱导的血管平滑肌细胞增殖,促进BACKGROUND:Irisin is released from fibronectin typeⅢdomain containing 5(FNDC5)after hydrolysis and modification.In addition to regulating the main physiological functions of glucolipid metabolism,irisin also has the ability to regulate the proliferation and apoptosis of a variety of cells.OBJECTIVE:To investigate the role and mechanism of irisin/peroxisome proliferator-activated receptor alpha(PPARα)signaling pathway mediated heme oxygenase-1 in regulating the balance between proliferation and apoptosis of vascular smooth muscle cells.METHODS:FNDC5 ove rexpression and silencing lentivirus vector were used to transfect vascular smooth muscle cells to increase or inhibit irisin production,and an in vitro cell model was constructed.There were seven groups:blank control group,overexpressed empty vector group,FNDC5 overexpression group,overexpressed empty vector+PPARαinhibitor(GW6471)group,FNDC5 overexpression+GW6471 group,silencing empty vector group,and sh-FNDC5 group.The mRNA and protein expression levels of FNDC5,PPARa,heme oxygenase-1,Bax,and Bcl-2 were detected by real-time PCR and western blot assay,respectively.Apoptosis of vascular smooth muscle cells was detected by flow cytometry and proliferation activity of vascular smooth muscle cells was detected by cell counting kit-8 assay.RESULTS AND CONCLUSION:(1)Compared with the blank control group and overexpressed empty vector group,the expressions of FNDC5,P PARa.heme oxygenase-1,and Bax were increased,the expressions of Bcl-2 were decreased,and the ratio of Bax/Bcl-2 was increased in the FNDC5 group and FNDC5+GW6471 group(all P<0.05).(2)Compared with the blank control group and silencing empty vector group,the expressions of FNDC5,P PARα,heme oxygenase-1,and Bax were decreased,the expressions of Bcl-2 were increased,and the ratio of Bax/Bcl-2 was decreased in the sh-FNDC5 group(all P<0.05).(3)Compared with the FNDC5 group,the expressions of P PARa,heme oxygenase-1,and Bax were significantly decreased,Bcl-2 expression was increased,and Bax/Bcl-2 ratio was

关 键 词：鸢尾素 过氧化物酶体增殖物激活受体Α 血红素氧合酶1 慢病毒 转染 血管平滑肌细胞 增殖 凋亡 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R543.1