泻肺利水合剂对异丙肾上腺素损伤大鼠心肌细胞线粒体钙超载的改善作用及机制  

Effect and mechanism of Xiefei Lishui Mixture on mitochondrial overload of myocardial cells injured by isoproterenol

作　　者：梁立新 魏鹏路 刘红旭[1] 刘子豪 来晓磊[1] LIANG Lixin;WEI Penglu;LIU Hongxu;LIU Zihao;LAI Xiaolei(Department of Cardiology,Beijing Hospital of Traditional Chinese Medicine,Capital Medical University,Beijing 100010,China)

机构地区：[1]首都医科大学附属北京中医医院心血管科,北京100010

出　　处：《北京中医药》2025年第2期149-156,共8页Beijing Journal of Traditional Chinese Medicine

基　　金：国家中医药管理局中医药循证能力建设项目(2019XZZX-XXG001);北京市医院管理局重点医学专业发展计划项目(ZYLX201817)。

摘　　要：目的研究泻肺利水合剂对异丙肾上腺素(isoproterenol,ISO)损伤大鼠心肌细胞(H9c2细胞)线粒体钙超载的改善作用及机制。方法体外培养H9c2细胞,取部分H9c2细胞随机分为正常组、模型组、中药组、卡托普利组,除正常组外,各组均先使用含ISO的培养基培养48 h,正常组、模型组用20%生理盐水含药血清、中药组用20%泻肺利水含药血清、卡托普利组用20%卡托普利含药血清培养基再培养48 h。取部分H9c2细胞随机分为转染对照组(si-NC组)、转染中药对照组(si-NC+中药组)、转染卡托普利对照组(si-NC+卡托普利组)、转染模型组(si-GRP75组)、转染中药模型组(si-GRP75+中药组)、转染卡托普利模型组(si-GRP75+卡托普利组),将转染后的各组细胞均先用含ISO的培养基培养48 h,si-NC组、si-GRP75组加入20%生理盐水含药血清培养基,si-NC+中药组、si-GRP75+中药组加入20%泻肺利水含药血清培养基,si-NC+卡托普利组、si-GRP75+卡托普利组加入20%卡托普利含药血清培养基,再培养48 h。CCK-8法检测细胞活力,荧光定量PCR检测肌醇-1,4,5三磷酸受体(IP3R)、葡萄糖调节蛋白75(GRP75)、电压依赖性阴离子通道蛋白1(VDAC1)、线粒体融合蛋白2(MFN2)mRNA表达,Western blotting法检测IP3R、GRP75、VDAC1、MFN2蛋白表达,Rhod2-AM Ca^(2+)荧光探针法检测线粒体内Ca^(2+)含量。结果与正常组比较,模型组IP3R、GRP75、VDAC1 mRNA、蛋白表达及线粒体Ca^(2+)含量高,MFN2 mRNA和蛋白表达低(P<0.05);与模型组比较,中药组、卡托普利组IP3R、GRP75、VDAC1 mRNA、蛋白表达及线粒体Ca^(2+)含量低,MFN2 mRNA和蛋白表达高(P<0.05);与si-NC组比较,si-NC+中药组、si-GRP75组细胞存活率高,IP3R、GRP75、VDAC1 mRNA和蛋白表达低,线粒体Ca^(2+)含量低,MFN2 mRNA和蛋白表达高(P<0.05);与si-NC+中药组比较,si-GRP75+中药组细胞存活率高,IP3R、GRP75、VDAC1 mRNA、蛋白表达及线粒体Ca^(2+)含量低,MFN2 mRNA、蛋白表达高(P<0.Objective To investigate the protective effect of Xiefei Lishui Mixture containing serum on isoproterenol(ISO)injured rat cardiomyocytes(H9c2)and its mechanism.Method H9c2 cells were cultured in vitro to construct a cell model of ISO injury.First,the cells were divided into 4 groups:normal group,model group,Chinese medicine group and captopril group.The GRP75 gene was knocked down by plasmid transfection,and then the cells were divided into 6 groups:si-NC group,si-NC+Chinese medicine group,si-NC+captopril group,si-GRP75 group,si-GRP75+Chinese medicine group and si-GRP75+captopril group,which were cultured with corresponding serum.The cell viability of each group was detected dy CCK-8 method,the mRNA levels of IP3R,GRP75,VDAC1 and MFN2 were detected dy real-time fluorescence quantitative PCR,the protein expressions of IP3R,GRP75,VDAC1 and MFN2 were detected dy Western blot,and the mitochondrial Ca^(2+)content was detected dy Rhod2-AM Ca^(2+)fluorescent prode.Result Compared to the normal group,the model group exhibited increased mRNA and protein expressions of IP3R,GRP75,VDAC1,and mitochondrial Ca^(2+)content(P<0.05).Conversely,the mRNA and protein expressions of MFN2 were decreased in the model group(P<0.05).In comparison to the model group,both traditional Chinese medicine and captopril groups showed reduced mRNA and protein expressions of IP3R,GRP75,VDAC1 as well as mitochondrial Ca^(2+)content;meanwhile,they exhibited increased mRNA and protein expressions of MFN2(P<0.05).Furthermore,compared to the si-NC group,both si-NC+traditional Chinese medicine group and si-GRP75 group demonstrated improved cell survival rate along with decreased levels of IP3R,GRP75 mRNA/protein expressions as well as mitochondrial Ca^(2+)content;additionally,they displayed increased levels of MFN2 mRNA/protein expressions(P<0.05).Notably,the si-GRP75+traditional Chinese medicine group had a higher cell survival rate than the si-NC+traditional Chinese medicine group;it also exhibited lower levels of IP3R/GRP75/VDAC1 mRNA/protein express

关 键 词：泻肺利水合剂 异丙肾上腺素 大鼠 心肌细胞 线粒体钙超载 

分 类 号：R285.5[医药卫生—中药学]