心肌细胞肥大时线粒体三磷酸腺苷敏感钾通道和线粒体自噬的变化  

Changes in mitochondrial adenosine triphosphate sensitive potassium channel and mitochondrial autophagy during cardiomyocyte hypertrophy

作　　者：程晓蔚 朱庆磊 Cheng Xiaowei;Zhu Qinglei(Medical School of Chinese PLA,Beijing 100853,China;Institute of Cardiological Diseases,Senior Department of Cardiology,Sixth Medical Center,Chinese PLA General Hospital,Beijing 100853,China)

机构地区：[1]解放军医学院,北京100853 [2]中国人民解放军总医院第六医学中心心血管病医学部心血管病研究所,北京100853

出　　处：《中华老年多器官疾病杂志》2023年第5期372-376,共5页Chinese Journal of Multiple Organ Diseases in the Elderly

基　　金：国家自然科学基金(81570349);军队重大项目(ALB18J002)。

摘　　要：目的 研究心肌细胞肥大时线粒体三磷酸腺苷(ATP)敏感钾通道亚基及线粒体自噬的变化。方法 选择出生24 h的大鼠乳鼠体外培养原代心肌细胞,将其随机分为2组。对照组细胞采用加含10%胎牛血清的高糖培养基进行培养,模型组细胞在含10%胎牛血清的高糖培养基中加入异丙肾上腺素(ISO)140μmol/L处理48 h。酶联免疫吸附法检测心肌细胞培养上清液氨基末端脑钠肽前体(NT-proBNP)浓度,实时荧光定量聚合酶链式反应检测心房钠尿肽(ANP)、脑利钠肽(BNP)、β-肌球蛋白重链(β-MHC)、内向整流钾通道6.1(Kir6.1)和自噬相关基因8(Atg8)的mRNA表达,蛋白免疫印迹法检测Kir6.1和线粒体自噬相关蛋白微管相关蛋白1轻链3(LC3)和FUN14结构域蛋白1(FUNDC1)的表达。采用GraphPad Prism v9.0.0软件进行数据分析。组间比较采用t检验。结果 经ISO处理后,模型组心肌细胞上清液NT-proBNP浓度[(1 699.43䥺SymbolqB@407.01)pg/ml]显著高于对照组[(808.68䥺SymbolqB@91.46)pg/ml],标志性肥大基因ANP、BNP和β-MHC的mRNA表达增加,Kir6.1和Atg8的mRNA表达显著降低,线粒体ATP敏感钾通道蛋白Kir6.1和线粒体自噬相关蛋白LC3,FUNDC1表达明显下降(均P<0.05)。结论 心肌细胞肥大时线粒体ATP敏感钾通道表达和线粒体自噬水平均降低。Objective To study the changes in mitochondrial adenosine triphosphate(ATP)sensitive potassium channel Kir6.1 and mitophagy in cardiomyocyte hypertrophy.Methods The primary cardiomyocytes derived from^(2)4-hour-old neonatal rats born were randomly divided into control group and model group.The control group was cultured in a high glucose medium containing 10%fetal bovine serum,while for the model group,140μmol/L isoproterenol(ISO)was added to the medium for 48 h to establish a cellular model of cardiomyocyte hypertrophy.The concentration of N-terminal pro brain natriuretic peptide(NT-proBNP)in the culture supernatant of cardiomyocytes was detected by enzyme linked immunosorbent assay.The mRNA expression of atrial natriuretic peptide(ANP)mRNA,brain natriuretic peptide(BNP)mRNA,β-myosin heavy chain(β-MHC)mRNA,inward rectifyimg potassium channel 6.1(Kir6.1)mRNA and autophagy related gene 8(Atg8)mRNA were detected by quantitative real-time polymerase chain reaction(qRT-PCR).The expression of Kir6.1,microtubule-associated protein 1 light chain 3(LC3)and FUN14 domain-containing protein 1(FUNDC1)were detected by Western blotting.GraphPad Prism statistics v9.0.0 was used for statistical analysis.Data comparison between two groups was performed using student′s t test.Results The supernatant concentration of NT-proBNP was significantly higher in the model group than the control group[(1699.430x0E䥺SymbolqB@0x0F407.01)vs(808.680x0E䥺SymbolqB@0x0F91.46)pg/ml,P<0.05].When compared with the control group,the model group had obviously increased ANP,BNP andβ-MHC expression and decreased Kir6.1 and Atg 8 expression at mRNA level,and reduced expression of Kir6.1 as well as LC3 and FUNDC1 at protein level(all P<0.05).Conclusion The expression of mitochondrial ATP sensitive potassium channel and the level of mitophagy are decreased in cardiomyocyte hypertrophy.

关 键 词：心肌细胞肥大 异丙肾上腺素 内向整流钾通道6.1 自噬相关基因8 微管相关蛋白1轻链3 FUN14结构域蛋白1 

分 类 号：R542.2[医药卫生—心血管疾病]