花旗松素调节AMPK/mTOR信号通路对急性心肌梗死大鼠心肌损伤的影响  

Impact of taxifolin on myocardial injury in rats with acute myocardial infarction by regulating AMPK/mTOR signal pathway

作　　者：兰惠琴 朱光旭 王志远 黎德才 LAN Huiqin;ZHU Guangxu;WANG Zhiyuan;LI Decai(Department of Laboratory Medicine,Chongqing Kanghua Zhonglian Cardiovascular Disease Hospital,Chongqing 400025,China;School of Basic Medical Sciences,Chengdu University of Traditional Chinese Medicine,Chengdu,Sichuan 610075,China)

机构地区：[1]重庆康华众联心血管病医院检验科,重庆400025 [2]成都中医药大学基础医学院,四川成都610075

出　　处：《安徽医药》2024年第12期2339-2344,共6页Anhui Medical and Pharmaceutical Journal

基　　金：四川省卫生健康科研课题普及项目(19PJ190)。

摘　　要：目的探讨花旗松素(TAX)调节AMP活化的蛋白质激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路对急性心肌梗死(AMI)大鼠心肌损伤的影响。方法2022年2—11月,从90只雄性SD大鼠中按随机数字表法选择18只大鼠作为空白对照(CON)组,其他大鼠构建AMI模型,将造模成功的AMI大鼠按随机数字表法分为AMI组、TAX组(50 mg/kg)、DOR组(0.01 mol/L的AMPK/mTOR抑制剂DOR)、TAX+DOR组(50 mg/kg TAX+0.01 mol/L的AMPK/mTOR抑制剂DOR),每组18只大鼠。观察超声心动图评估舒张期室间隔厚度(IVSd)、舒张期左室内径(LVIDd)、收缩期左室内径(LVIDs)、左心室舒张期后壁厚度(LVPWd)、左室射血分数(EF)和缩短分数(FS);酶联免疫吸附测定(ELISA)检测心肌损伤标志物肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)以及炎症因子白细胞介素(IL)-1β水平;苏木精-伊红(HE)染色检测心脏组织病理变化;2,3,5-氯化三苯基四氮唑(TTC)染色检测心肌梗死面积;TUNEL染色检测细胞凋亡;蛋白质印迹法检测AMPK/mTOR信号通路蛋白水平。结果CON组心脏组织结构完整,细胞排列整齐有序,染色清晰;AMI组出现炎症细胞浸润现象;TAX组炎症细胞浸润现象减轻,而DOR组炎症细胞浸润现象加重;TAX+DOR组与AMI组结构相似。与CON组相比,AMI组LVIDd、LVIDs、CK、CK-MB、LDH、IL-1β含量、细胞凋亡率、梗死面积[(17.82±2.31)%比(3.91±0.15)%]、mTOR(1.26±0.14比0.54±0.04)、核苷酸结合结构域富含亮氨酸重复序列和含热蛋白结构域受体3(NLRP3)(0.76±0.08比0.26±0.02)蛋白水平显著升高(P<0.05),IVSd、LVPWd、EF、FS以及p-AMPK/AMPK(0.49±0.05比0.93±0.07)蛋白水平显著下降(P<0.05);与AMI组相比,TAX组LVIDd、LVIDs、CK、CK-MB、LDH、IL-1β含量、细胞凋亡率、梗死面积[(12.36±1.76)%比(17.82±2.31)%]、mTOR(0.69±0.07比1.26±0.14)、NLRP3(0.39±0.04比0.76±0.08)蛋白水平显著降低(P<0.05),IVSd、LVPWd、EF、FS以及p-AMPK/AMPK(0.85±0.09比0.49±0.0Objective To investigate the impact of taxifolin(TAX)on myocardial injury in rats with acute myocardial infarction(AMI)by regulating AMP-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)signal pathway.Methods From February 2022 to November 2022,18 male SD rats were selected from 90 ones as blank control group(CON group)according to random number table method,and AMI models were constructed for other rats.AMI rats successfully modeled were assigned into AMI group,TAX group(50 mg/kg),DOR group(0.01 mol/L AMPK/mTOR inhibitor DOR),TAX+DOR group(50 mg/kg TAX+0.01 mol/L AMPK/mTOR inhibitor DOR)by random number table method,with 18 rats in each group.Diastolic inlet ventricular septal thickness(IVSd),diastolic left ventricular internal diameter(LVIDd),systolic left ventricular internal diameter(LVIDs),left ventricular posterior diastolic wall thickness(LVPWd),left ventricular ejection fraction(EF)and fractional shortening(FS)were evaluated by echocardiography.The levels of myocardial injury markers creatine kinase(CK),creatine kinase isoenzyme(CK-MB),lactate dehydrogenase(LDH)and inflammatory factors interleukin1β(IL-1β)were detected by enzyme-linked immunosorbent assay(ELISA).Hematoxylin eosin(HE)staining was used to detect the pathological changes of heart tissue,and myocardial infarction area was measured by 2,3,5,triphenyl-2Htetrazolium chloride(TTC)staining.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining was used to detect apoptosis,and Western blotting was used to detect the protein level of AMPK/mTOR signal pathway.Results In CON group,the heart tissue structure was complete,the cells were orderly arranged,and the staining was clear;inflammatory cell infiltration occurred in AMI group;inflammatory cell infiltration was reduced in TAX group,but increased in DOR group;the structure of TAX+DOR group was similar to that of AMI group.Compared with CON group,the levels of LVIDd,LVIDs,CK,CK-MB,LDH,IL-1β,apoptosis rate,infarct size[(17.82±2.31)%vs.(3.91±0.15)%],l

关 键 词：黄杉属 花旗松素 心肌梗死 AMP活化的蛋白质激酶/哺乳动物雷帕霉素靶蛋白信号通路 心肌损伤 炎症 

分 类 号：R285.5[医药卫生—中药学]