MIF-AMPK调控能量代谢介导七氟醚后处理抗心肌缺氧复氧损伤的机制研究  

Mechanism of energy metabolism mediated by MIF-AMPK regulates on sevoflurane postconditioning against myocardial hypoxia-reoxygenation injury

作　　者：努尔比艳·克尤木[1] 曹浩然 何雨璇 辛铭秀 马海平[1] Nuerbiyan Keyumu;CAO Haoran;HE Yuxuan;MA Haiping(The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学第一附属医院,乌鲁木齐830054

出　　处：《新疆医科大学学报》2022年第9期958-963,共6页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区围术期器官保护重点实验室项目(2021D04021)。

摘　　要：目的探索巨噬细胞移动抑制因子介导的腺苷酸活化蛋白激酶(MIF-AMPK)信号通路在调控七氟醚后处理(SPostC)对抗心肌缺氧复氧(H/R)损伤中的作用机制,为临床对抗心肌缺血再灌注损伤提供理论依据。方法实验采用H9C2心肌细胞建立H/R模型,将心肌细胞随机分为四组:空白对照组(Control组)、缺氧/复氧组(H/R组)、SPostC组(H/R+SPostC组)、MIF抑制剂组(H/R+SPostC+IOS-1组)。SPostC是在细胞开始复氧时应用2.4%七氟醚进行处理15分钟。各组细胞存活率的检测采用CCK-8比色法;MIF的含量采用ELISA试剂盒检测;二氢乙啶(DHE)染色法检测细胞内活性氧(ROS)含量;采用检测试剂盒测细胞内二磷酸腺苷(ADP)、三磷酸腺苷(ATP)、乙酰辅酶A(AcCoA)的含量;Western blotting检测AMPK蛋白表达水平。结果(1)H/R+SPostC组与H/R组比较细胞存活率明显增高(P<0.01),而与H/R+SPostC+IOS-1组比细胞较存活率降低(P<0.01);(2)与Control组比较,其余3组MIF表达水平显著增高(P<0.05),而与H/R+SPostC组比较,H/R+SPostC+IOS-1组MIF含量显著降低(P<0.01);(3)AMPKα的mRNA和蛋白表达水平在4组间差异无统计学意义,与H/R组比较,H/R+SPostC组p-AMPKα蛋白表达水平显著增高,而H/R+SPostC+IOS-1的表达明显降低(P<0.05);(4)与对照组相相比,其余3组ADP、ATP含量均下降,H/R+SPostC组较H/R组显著增高,而H/R+SPostC+IOS-1组显著下降。与对照组比较,H/R组和H/R+SPostC+IOS-1组AcCoA含量明显增加;与H/R组比较,H/R+SPostC组和H/R+SPostC+IOS-1组AcCoA含量明显减少,H/R+SPostC+IOS-1组较H/R+SPostC组AcCoA含量显著增加,差异均有统计学意义(P<0.05);(5)与对照组相比,其余3组ROS含量均增高(P<0.05);与H/R组比较,H/R+SPostC组ROS含量明显减少(P<0.05)。结论SPostC对抗心肌H/R损伤中通过增加MIF的表达,改善AMPK的磷酸化水平,增加ADP、ATP的合成并减少AcCoA堆积,促进心肌胞能量合成,降低ROS生成,从而对抗心肌细胞H/R损伤。Objective To explore the mechanism of adenylate activated protein kinase(MIF-AMPK)signaling pathway mediated by macrophage migration inhibitor in regulating sevoflurane post-treatment SPostC)against myocardial hypoxia/reoxygenation(H/R)injury,and to provide theoretical basis for clinical treatment against myocardial ischemia/reperfusion injury.Methods H9c2 cardiomyocytes were used to establish hypoxia/reoxygenation(H/R)model.The cardiomyocytes were randomly divided into four groups:blank control group(Control group),hypoxia/reoxygenation group(H/R group),SPostC group(H/R+SPostC group),MIF inhibitor group(H/R+SPostC+IOS-1 group).SpostC was treated with 2.4%sevoflurane for 15 minutes when the cells began to reoxygenate.CCK-8 colorimetric method was used to detect the cell survival rate of each group;MIF was detected by ELISA kit;Dihydroethidine(DHE)staining was used to detect the content of intracellular reactive oxygen species(ROS);The contents of adenosine diphosphate(ADP),adenosine triphosphate(ATP)and acetyl coenzyme A(AcCoA)in the cells were measured by the detection kit;AMPK protein expression was detected by Western blotting.Results(1)The cell survival rate of H/R+SPostC group was significantly higher than that of H/R group(P<0.01),but itlower than that of H/R+SPostC+ios-1 group(P<0.01);(2)Compared withcontrol group,the expression level of MIF in the other three groups weresignificantly higher(P<0.05),while compared with H/R+SPostC group,the content of MIF inH/R+SPostC+IOS-1 group was significantly lower(P<0.01);(3)There was no significant difference inmRNA and protein expression levels of AMPKαamong the four groups,but compared with H/R group,the expression level of p-AMPKαprotein in H/R+SPostC group was significantly higher,while the expression level of H/R+SPostC+IOS-1 was significantly lower(P<0.05);(4)Compared with Control group,the contents of ADP and ATP in the other three groups were decreased;H/R+SPostC group was significantly higher than H/R group,while H/R+SPostC+IOS-1 group was significantly lo

关 键 词：七氟醚后处理 心肌细胞 巨噬细胞移动抑制因子 腺苷酸活化激活蛋白 

分 类 号：R551.2[医药卫生—血液循环系统疾病]