丝裂原活化蛋白激酶/细胞外信号调节激酶信号通路在穿心莲内酯治疗心肌缺血/再灌注损伤大鼠中的分子机制  

The molecular mechanism of MAPK/ERK1/2 signaling pathway in myocardial ischemia/reperfusion injury rats treated by andrographolide

作　　者：田俊斌[1] 赵静[1] 吕建瑞[1] 罗斌[1] 马磊[1] TIAN Junbin;ZHAO Jing;LYU Jianrui;LUO Bin;MA Lei(The Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004,China)

机构地区：[1]西安交通大学第二附属医院麻醉科,陕西西安710004

出　　处：《陕西中医》2022年第5期575-579,共5页Shaanxi Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金资助项目(81601148)。

摘　　要：目的:从丝裂原活化蛋白激酶/细胞外信号调节激酶(MAPK/ERK1/2)信号通路研究穿心莲内酯(AG)对心肌缺血/再灌注损伤(MI/RI)大鼠的分子机制,为AG能够安全地用于MI/RI的预防提供参考。方法:雄性SPF级SD大鼠60只,购入后先全部予适应性喂养2周,后将其随机分为假手术组、模型组、银杏内酯B干预组,AG治疗组(低、中和高三个剂量组),每组10只,分组后于次日给予每个组别相应药物干预14 d,14 d后采用经典法构建MI/RI模型(心肌缺血30 min,再灌注120 min),随后采集血液检测乳酸脱氢酶(LDH)、肌酸激酶同工酶(CK-MB)和肌钙蛋白I(cTnI)含量,同时采集心肌,检测心肌组织病理学、心肌Ca^(2+)-Mg^(2+)-ATP活力、Na^(+)-K^(+)-ATP活力、丙二醛(MDA)含量和超氧化物歧化酶(SOD)含量,最后采用免疫印迹法检测心肌丝裂原活化蛋白激酶p38(p38-MAPK)和细胞外调节蛋白激酶(p-ERK1/2)蛋白表达特点。结果:组织病理学表现为假手术组心肌细胞排列整齐、细胞呈长纤维状,模型组心肌纤维则出现明显且大量的断裂,与模型组相比,各治疗组均有不同程度好转;与假手术组比较,模型组大鼠血清LDH、CK-MB、cTnI、心肌MDA和心肌p38-MAPK蛋白表达均明显升高(P<0.05)、心肌SOD、Ca^(2+)-Mg^(2+)-ATP、Na^(+)-K^(+)-ATP、心肌p-ERK1/2蛋白表达均明显降低(P<0.05);与模型组相比,银杏内酯B组和AG低、中和高剂量组血清LDH、CK-MB、cTnI、心肌MDA以及心肌p38-MAPK蛋白表达均明显降低(P<0.05),心肌SOD、Ca^(2+)-Mg^(2+)-ATP、Na^(+)-K^(+)-ATP和心肌p-ERK1/2蛋白表达均明显升高(P<0.05)。结论:AG预防给药,能够有效的保护MI/RI大鼠心肌及心功能,其作用机制可能与AG能够调节MAPK/ERK1/2信号通路有关。Objective:Study on the therapeutic effect and mechanism of andrographolide(AG)on myocardial ischemia/reperfusion injury(MI/RI)in rats from MAPK/ERK1/2 signaling pathway.Methods:60 male clean SD rats were fed adaptively for 2 weeks after purchase,then they were randomly divided into sham operation group,model group,ginkgolide B intervention group,and andrographolide treatment group(low,middle,and high dose groups).After grouping,the corresponding drugs were given for 14 days and 14 days later,the MI/RI model was established by routine method(myocardial ischemia 30 min,reperfusion 120 min).Then blood samples were collected to detect the contents of lactate dehydrogenase(LDH),creatine kinase isoenzyme(CK-MB),and troponin I(cTnI).At the same time,myocardial histopathology,myocardial Ca^(2+)-Mg^(2+)-ATP activity,Na^(+)-K^(+)-ATP activity,malondialdehyde(MDA)content,and superoxide dismutase(SOD)content were measured.Finally,the expression of mitogen-activated protein kinase p38(p38-MAPK)and extracellular regulated protein kinase(p-ERK1/2)in the myocardium was detected by Western blot.Results:Myocardial HE staining showed that the myocardial cells in the sham operation group were uniform and arranged neatly.The myocardial fibers in the model group were extensively exercised,compared with the model group,the cells in the three treatment groups were significantly improved.Compared with the sham operation group,the model group serum LDH,CK-MB,cTnI,myocardial MDA,and p38-MAPK protein expression were significantly increased(P<0.05),myocardial SOD,Ca^(2+)-Mg^(2+)-ATP,Na^(+)-K^(+)-ATP,myocardial p-ERK1/2 protein expression were significantly decreased(P<0.05),compared with the model group,ginkgolides B group and andrographolide low,medium and high dose group for serum LDH,CK-MB,cTnI,myocardial MDA,and the protein expression of p38-MAPK was significantly decreased(P<0.05),and the SOD,Ca^(2+)-Mg^(2+)-ATP,Na^(+)-K^(+)-ATP,myocardial p-ERK1/2 protein were significantly increased(P<0.05).Conclusion:Andrographolide can effectively p

关 键 词：心肌缺血/再灌注损伤 穿心莲内酯 MAPK/ERK1/2信号通路 p38-MAPK蛋白 p-ERK1/2蛋白 大鼠 

分 类 号：R542.22[医药卫生—心血管疾病]