北京中医药2025，Vol.44Issue(2)：149-156,8.DOI:10.16025/j.1674-1307.2025.02.007
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
摘要
Abstract
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 Ca2+content was detected dy Rhod2-AM Ca2+fluorescent prode.Result Compared to the normal group,the model group exhibited increased mRNA and protein expressions of IP3R,GRP75,VDAC1,and mitochondrial Ca2+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 Ca2+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 Ca2+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 expressions along with reduced mitochondrial Ca2+content while showing elevated levels of MFN2 mRNA/protein expressions(P<0.05).No significant differences were observed in terms of cell survival rate or mitochondrial Ca2+content among the si-GRP75 group,si-GRP75+traditional Chinese medicine group or si-GRP75+captopril group(P>0.05).Conclusion Xiefei Lishui Mixture can improve the survival rate of ISO injured rat H9c2 cells and reduce mitochondrial Ca2+overload,and its mechanism may relate to down regulation the IP3R/GRP75/VDAC1 pathway.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Key words
Xiefei Lishui Mixture/ISO/rats/H9c2 cells/mitochondrial overload引用本文复制引用
梁立新,魏鹏路,刘红旭,刘子豪,来晓磊..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[J].北京中医药,2025,44(2):149-156,8.基金项目
国家中医药管理局中医药循证能力建设项目(2019XZZX-XXG001) （2019XZZX-XXG001）
北京市医院管理局重点医学专业发展计划项目(ZYLX201817) （ZYLX201817）
