摘要
目的探讨烟酰胺单核苷酸腺苷酸转移酶2(Nmnat2)对异丙肾上腺素(ISO)诱导的肥大心肌细胞糖酵解的影响。方法将H9C2心肌细胞分为四组,均加入空白血清预处理12h。对照组转染短发夹RNA(shRNA)-对照质粒48h;模型组加入ISO 10μmol/L孵育48h,构建肥大心肌细胞模型;干扰组转染shRNA-Nmnat2质粒48h;模型+干扰组加入ISO 10μmol/L孵育48h,随后转染shRNA-Nmnat2质粒48h。比较各组细胞活力以及Nmnat2、α-肌球蛋白重链(α-MHC)、β-MHC、NF-κB mRNA和蛋白表达,检测各组葡萄糖摄取量以及乳酸、丙酮酸激酶(PK)、己糖激酶(HK)和磷酸果糖激酶(PFK)含量。结果第5天,与对照组比较,模型组和干扰组细胞活力降低(P<0.05);与模型组比较,模型+干扰组细胞活力降低(P<0.05)。与对照组比较,模型组和干扰组细胞Nmnat2和α-MHC mRNA和蛋白表达降低,β-MHC和NF-κB mRNA和蛋白表达增加,葡萄糖摄取量以及乳酸、PK、HK和PFK含量升高(P<0.05);与模型组比较,模型+干扰组细胞Nmnat2和α-MHC mRNA和蛋白表达降低,β-MHC和NF-κB mRNA和蛋白表达增加,葡萄糖摄取量以及乳酸、PK、HK和PFK含量升高(P<0.05)。结论干扰Nmnat2能够促进ISO诱导的心肌细胞肥大,增加心肌细胞的糖酵解。
Objective To explore the effect of nicotinic acid mononucleotide adenylyltransferase 2(Nmnat2)on the glycolysis of hypertrophic cardiomyocytes induced by isoproterenol.Methods H9C2 cardiomyocytes were divided into four groups and pretreated with blank serum for 12 hours.Group A was transfected with shRNA-control plasmid for 48 hours.Group B was incubated with isoproterenol 10μmol/L for 48 hours to construct the hypertrophic cardiomyocyte model.Group C was transfected with the shRNA-Nmnat2 plasmid for 48 hours.Group D was incubated with isoproterenol 10μmol/L for 48 hours,and then transfected with shRNA-Nmnat2 plasmid for 48 hours.The cell viability and the mRNA and protein expressions of Nmnat2,α-myosin heavy chain(α-MHC),β-MHC and NF-κB were compared among the four groups.The glucose uptake content and the contents of lactic acid,pyruvate kinase(PK),heptokinase(HK)and phosphofructokinase(PFK)were detected.Results Compared with group A,the cell viability in groups of B and C was decreased on the 5th day of treatment(P<0.05).Compared with group B,the cell viability was decreased in group D(P<0.05).Compared with group A,the mRNA and protein expressions of Nmnat2 andα-MHC were decreased,while those ofβ-MHC and NF-κB were increased,the glucose uptake content and the contents of lactic acid,PK,HK and PFK were increased in groups of B and C(P<0.05).Compared with group B,the mRNA and protein expressions of Nmnat2 andα-MHC were decreased,while those ofβ-MHC and NF-κB were increased,the glucose uptake content and the contents of lactic acid,PK,HK and PFK were increased in group D(P<0.05).Conclusion Interference with Nmnat2 can promote isoproterenol-induced cardiomyocyte hypertrophy and increase the glycolysis of cardiomyocytes.
作者
裴菱花
陈静文
柳浦青
PEI Linghua;CHEN Jingwen;LIU Puqing(Department of Cardiovascular Medicine,Affiliated Second Hospital,Zhejiang Chinese Medical University,Hangzhou 310005,CHINA)
出处
《江苏医药》
2025年第6期541-545,共5页
Jiangsu Medical Journal
基金
浙江省自然科学基金项目(LYY22H280001)
浙江省中医药科技计划项目(2023ZL453)。
