基于AMPK/mTOR信号通路探讨麦粒灸预处理对心肌缺血再灌注损伤大鼠心肌细胞中自噬及凋亡基因caspase-3表达的影响  

The effects of wheat moxibustion pretreatment on the expression of autophagy and apoptosis gene caspase-3 in myocardial cells of rats with myocardial ischemia-reperfusion injury based on AMPK/mTOR signaling pathway

作　　者：宋万珍 刘顺畅 潘玉璟 刘琼琼 丁雅萍 张宏如[1] 顾一煌[1] SONG Wan-Zhen;LIU Shun-Chang;PAN Yu-Jing(College Acupuncture,Massage,Health and Rehabilitation,Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu,China)

机构地区：[1]南京中医药大学针灸推拿·养生康复学院,江苏南京210023 [2]南京中医药大学针药结合教育部重点实验室,江苏南京210023

出　　处：《中国老年学杂志》2024年第14期3435-3441,共7页Chinese Journal of Gerontology

基　　金：国家自然科学基金面上项目(81974583);江苏省高校护理学优势学科建设工程资助项目(2019YSHL051);南京中医药大学校自然科学基金青年项目(NZY81704169)。

摘　　要：目的 麦粒灸预处理观察心肌缺血再灌注损伤(MIRI)大鼠心肌组织中细胞自噬和凋亡情况的影响,探讨麦粒灸预处理抗MIRI的可能机制。方法 将成年雄性SD大鼠随机分为假手术组、模型(MI)组、麦粒灸组、抑制剂组。MI组、麦粒灸组和抑制剂组采用冠状动脉左前降支(LAD)结扎术建立MIRI,假手术组仅在LAD下方穿线不结扎。麦粒灸组、抑制剂组于造模前麦粒灸双侧“内关”穴,每穴灸5柱,1次/d,连续干预7 d。采用生理信号采集系统记录心电图,观察术后30 min标准肢体Ⅱ导联ST段电位偏移情况;超声心动图评估术后24 h大鼠左心室射血分数(LVEF);酶联免疫吸附试验(ELISA)法检测血清中肌钙蛋白(cTnT)、肌酸激酶同工酶(CK-MB)含量;Western印迹检测AMP激活蛋白激酶(AMPK),p-AMPK,哺乳动物雷帕霉素蛋白(mTOR),p-mTOR,胱天蛋白酶(caspase)-3蛋白表达;实时荧光定量聚合酶链反应(PCR)观察caspase-3 mRNA表达水平。结果 与假手术组比较,MI组LVEF值、p-mTOR/mTOR蛋白表达显著降低,p-AMPK/AMPK蛋白及caspase-3蛋白及mRNA表达显著升高(P<0.01,P<0.001)。与MI组比较,麦粒灸组LVEF值及p-mTOR/mTOR蛋白水平明显上升,p-AMPK/AMPK蛋白、cTnT、CK-MB值及caspase-3蛋白及mRNA表达显著下降(P<0.01,P<0.001)。与麦粒灸组相比,抑制剂组CK-MB、cTnT值、caspase-3 mRNA及蛋白表达明显上升,LVEF值明显降低(P<0.01,P<0.001)。结论 麦粒灸预处理通过AMPK/mTOR通路可以有效减轻MIRI大鼠心肌组织的细胞自噬和凋亡水平,降低心肌梗死程度,实现心肌保护效应。Objective To observe the effects of wheat moxibustion on autophagy and apoptosis in myocardial tissue of myocar-dial ischemia reperfusion injury(MIRI)rats,and to explore the possible mechanism of anti-MIRI induced by wheat moxibustion.Methods Adult male SD rats were randomly divided into sham operation group,model(MI)group,wheat moxibustion group and in-hibitor group.MIRI was established by ligation of the left anterior descending branch of coronary artery(LAD)in MI group,wheat moxibustion group and inhibitor group,sutures were only threaded below the LAD without ligation in the sham operation group.The wheat moxibustion group and the inhibitor group were treated at both"Neiguan"points of wheat moxibustion,5 columns of moxibustion at each point,once a day,for 7 consecutive days.Physiological signal acquisition system was used to record electrocardiogram,and ST segment potential offset of leadⅡof standard limb was observed 30 min after operation.Left ventricular ejection fraction(LVEF)was evaluated 24 h after operation by echocardiography.The contents of serum troponin(cTnT)and creatine kinase isoenzyme(CK-MB)were detected by enzyme-linked immunosorbent assay(ELISA).AMP-activated protein kinase(AMPK),p-AMPK,mammalian rapa-mycin protein(mTOR),p-mTOR,caspase-3 protein expressions were detected by Western blotting.Real-time fluorescence quantitative polymerase chain reaction(PCR)was used to observe the mRNA expression level of caspase-3.Results Compared with sham opera-tion group,LVEF value and p-mTOR/mTORprotein expression were significantly decreased in MI group,p-AMPK/AMPK protein and caspase-3 protein and mRNA expressions were significantly increased(P<0.01,P<0.001).Compared with MI group,the LVEF value and p-mTOR/mTORprotein level were significantly increased,p-AMPK/AMPK protein,cTnT,CK-MB value and caspase-3 protein and mRNA expressions were significantly decreased in wheat moxibustion group(P<0.01,P<0.001).Compared with wheat moxibus-tion group,CK-MB,cTnT value,caspase-3 mRNA and protein expressions were sig

关 键 词：麦粒灸预处理 心肌缺血再灌注损伤 AMP激活蛋白激酶(AMPK) 哺乳动物雷帕霉素蛋白(mTOR) 胱天蛋白酶(caspase)-3 细胞凋亡 自噬 

分 类 号：R542.2[医药卫生—心血管疾病]