阿昔莫司对高糖诱导心肌细胞损伤保护作用机制研究  

Study on the protective effects and action mechanism of acipimox on cardiomyocyte injury induced by high glucose

作　　者：解园星[1] 王文锋[1] 田红燕[2] XIE Yuanxing;WANG Wenfeng;TIAN Hongyan(Department of Cardiology,Shangluo Central Hospital,Shaanxi,Shangluo 726000,China;不详)

机构地区：[1]陕西省商洛市中心医院心血管内科,726000 [2]西安交通大学第一附属医院周围血管科

出　　处：《河北医药》2022年第14期2096-2101,共6页Hebei Medical Journal

摘　　要：目的探讨阿昔莫司对高糖诱导的心肌细胞氧化应激损伤的影响,探讨其保护作用机制。方法取大鼠心肌H9C2细胞,分为对照组、低糖组、等渗组、高糖组,采用MTT法检测各组细胞存活率,Western blot法检测超氧化物歧化酶(SOD)及脂蛋白脂肪酶(LPL)表达水平。在高糖(30 mmol/L)条件下,用不同浓度阿昔莫司(0、5、10、25、50μg/L)干预处理培养24 h,检测细胞存活力及SOD、LPL水平。敲低SOD,并分为空白组、高糖组、阿昔莫司组(25μg/L)、Si-SOD组(转染SOD低表达序列腺病毒载体)、Si-NC组(转染不含Si-SOD序列的腺病毒空载体)、阿昔莫司+Si-SOD组;MMT法检测细胞存活率;透射电镜检测细胞超微结构;油红O染色观察脂质堆积;免疫荧光检测活性氧簇(ROS)蓄积;TUNEL法检测细胞凋亡;Western blot法测定LPL、抗氧化酶-超氧化物歧化酶2(SOD2)、脂质过氧化物酶体增殖物激活受体α(PPARα)、谷胱甘肽过氧化物酶(GPx)、过氧化氢酶(CAT)、心肌细胞脂质代谢相关指标-细胞膜糖蛋白(CD36)、葡萄糖转运蛋白4(GLUT4)蛋白表达。结果糖浓度越高,心肌细胞存活率及SOD蛋白表达越低,LPL表达越高(P<0.05)。高糖条件下,阿昔莫司干预浓度越高,心肌细胞存活率及SOD蛋白表达越高,LPL表达越低(P<0.05)。敲低SOD后,细胞脂质堆积、ROS蓄积、氧化应激损伤及凋亡越严重,PPARα介导的脂代谢异常途径活化越高,SOD抗氧化途径受到抑制(P<0.05)。阿昔莫司可提高高糖条件下细胞存活率,缓解氧化应激损伤及脂质堆积,抑制PPARα活化,并促进SOD抗氧化途径激活(P<0.05),但其作用可被Si-SOD逆转(P<0.05)。结论阿昔莫司通过抑制PPARα活化,升高SOD、GPx、CAT抗氧化相关蛋白表达,缓解脂质堆积及氧化应激损伤,对高糖诱导条件下心肌细胞起保护作用。Objective To investigate the protective effects and action mechanism of acipimox on cardiomyocyte injury induced by high glucose.Methods The ardiac muscle H9C2 cells of rats were divided into control group,low glucose group,isotonic group and high glucose group.MTT method was used to detect cell survival rate,Western Blot was used to detect the expression levels of superoxide dismutase(SOD)and lipoprotein lipase(LPL).Under high glucose(30mmol/L)condition,the cells were cultured and intervened by different concentrations of acipimox(0,5,10,25,50μg/L)for 24 hours,then the cell viability,and levels of SOD and LPL were detected.After knocking down SOD levels,the cells were divided into blank group,high glucose group,acipimox group(25μg/L),Si-SOD group(transfected with adenovirus vector with SOD low expression sequence),Si-NC group(transfected with adenovirus empty vector without Si-SOD sequence),acipimox+Si-SOD group.MMT was used to detect cell survival rate;transmission electron microscopy detected cell ultrastructure;oil red O staining was used to observe lipid accumulation;immunofluorescence detected reactive oxygen species(ROS)accumulation;TUNEL detected cell apoptosis;Western Blot measured the expression levels of LPL,antioxidant enzyme-superoxide dismutase 2(SOD2),lipid peroxisome proliferator activated receptorα(PPARα),glutathione peroxidase(GPx),catalase(CAT),cardiomyocyte lipid metabolism related indicators-cell membrane glycoprotein and glucose transporter 4.Results The higher the glucose concentration,the lower the survival rate of cardiomyocytes and the expression of SOD protein,and the higher the expression of LPL(P<0.05).Under high glucose conditions,the higher the intervention concentration of acipimox,the higher the survival rate of cardiomyocytes and the expression of SOD protein,and the lower the expression of LPL(P<0.05).After knocking down SOD,the cell lipid accumulation,ROS accumulation,oxidative stress damage and apoptosis were more serious,the activation of the abnormal lipid metabolism pat

关 键 词：阿昔莫司 心肌细胞 细胞存活率 高糖 

分 类 号：R542.2[医药卫生—心血管疾病]