柠檬酸盐对缺氧/复氧诱导的心肌细胞损伤及LKB1/AMPK信号通路的影响  

The Effects of Citrate on Hypoxia/Reoxygenation-Induced Myocardial Cell Injury and LKB1/AMPK Signal Pathway

作　　者：张博[1] 周学才[1] 朱冬梅 万翔[1] ZHANG Bo;ZHOU Xuecai;ZHU Dongmei;WAN Xiang(Xiantao First People's Hospital,Xiantao 433000,Hubei,China)

机构地区：[1]长江大学附属仙桃市第一人民医院,湖北仙桃433000

出　　处：《中西医结合心脑血管病杂志》2023年第10期1782-1787,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：湖北省卫生计生委科研项目(No.WJ2016-Y-36)。

摘　　要：目的:探讨柠檬酸盐对缺氧/复氧(H/R)诱导的心肌细胞损伤及肝激酶B1(LKB1)/磷酸腺苷活化蛋白激酶(AMPK)信号通路的影响。方法:将人心肌细胞(HCM)随机分为对照组(常规培养的HCM细胞)、H/R组(缺氧处理2.5 h,复氧处理2 h)、柠檬酸盐低、中、高剂量组(20μmol/mL、40μmol/mL、80μmol/mL柠檬酸盐培养液)、柠檬酸盐高剂量+抑制剂组(3 mg/kg LKB1/AMPK通路抑制剂Doxorubicin HCI、80μmol/mL柠檬酸盐培养液)。采用细胞增殖毒性检测(CCK-8)法测定各组HCM细胞增殖抑制率,生化法测定各组HCM细胞培养液中心肌损伤标志物肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)水平,流式细胞仪检测各组HCM细胞凋亡率,蛋白免疫印迹(Western Bloting)法测定HCM中凋亡相关蛋白(Bax、Bcl-2)及LKB1/AMPK通路相关蛋白的表达水平。结果:与对照组相比,H/R组HCM的细胞增殖抑制率、细胞凋亡率、CK-MB、LDH及Bax蛋白表达水平明显升高(P<0.05),Bcl-2蛋白表达水平明显降低(P<0.05)。与H/R组相比,柠檬酸盐低、中、高剂量组HCM的细胞增殖抑制率、细胞凋亡率、CK-MB、LDH及Bax蛋白表达水平明显降低(P<0.05),Bcl-2蛋白表达水平明显升高(P<0.05)。与柠檬酸盐高剂量组相比,柠檬酸盐高剂量+抑制剂组HCM的细胞增殖抑制率、细胞凋亡率、CK-MB、LDH及Bax蛋白表达水平明显升高(P<0.05),Bcl-2蛋白表达水平明显降低(P<0.05)。结论:柠檬酸盐可以促进受损的HCM细胞生存,可能与激活LKB1/AMPK信号通路有关。Objective:To investigate the effects of citrate on hypoxia/reoxygenation-induced myocardial cell injury and liver kinase B1(LKB1)/AMP-activated protein kinase(AMPK)signaling pathway.Methods:Human myocardial cells HCM were randomly divided into control group(conventionally cultured HCM cells),hypoxia/reoxygenation(H/R)group(hypoxia treatment for 2.5 hours,reoxygenation treatment for 2 hours),citrate low,medium,and high-dose groups(20μmol/mL,40μmol/mL,and 80μmol/mL citrate culture solution,respectively),citrate high-dose+inhibitor group(3 mg/kg LKB1/AMPK pathway inhibitor Doxorubicin HCI,80μmol/mL citrate culture solution).CCK-8 method was used to determine the inhibition rate of HCM cell proliferation in each group.Biochemical methods were used to determine the myocardial injury markers creatine kinase isoenzyme(CK-MB)and lactate dehydrogenase(LDH)in the culture medium of HCM cells in each group.Flow cytometry was used to detect the apoptosis rate of HCM cells in each group.Western Bloting method was used to determine the expression levels of apoptosis-related proteins(Bax,Bc-l 2)and LKB1/AMPK pathway related proteins in HCM cells.Results:Compared with control group,the cell proliferation inhibition rate,apoptosis rate,and the expression levels of CK-MB,LDH,and Bax proteins in HCM cells in H/R group were significantly increased(P<0.05),and the expression level of Bc-l 2 protein was significantly reduced(P<0.05).Compared with H/R group,the cell proliferation inhibition rate,apoptosis rate,and the expression levels of CK-MB,LDH,and Bax proteins in HCM cells in citrate low,medium,and high-dose groups were decreased(P<0.05),and the expression level of Bc-l 2 protein was decreased(P<0.05).Compared with citrate high-dose group,the cell proliferation inhibition rate,apoptosis rate,and the expression levels of CK-MB,LDH,and Bax proteins in HCM cells in citrate high-dose+inhibitor group were significantly increased(P<0.05),and the expression level of Bc-l 2 protein was significantly decreased(P<0.05).Conclusion:Citrate

关 键 词：心肌细胞损伤 柠檬酸盐 肝激酶B1/磷酸腺苷活化蛋白激酶 实验研究 

分 类 号：R542.2[医药卫生—心血管疾病]