芍药苷预处理激活AMPKα对在体大鼠心肌缺血再灌注损伤的保护作用

Protective effects of pretreatment with paeoniflorin on the myocardial ischemia reperfusion injury of rats in vivo by activating AMPKα

目的探究芍药苷预处理激活AMPKα信号通路对大鼠在体心肌缺血再灌注损伤的保护作用。方法本实验共用SPF级雄性SD大鼠30只,分三组,分别为假手术组、缺血再灌注组、药物预处理组,采用结扎SD大鼠心脏LAD的方法建立在体大鼠心肌缺血再灌注损伤模型,利用0.5%Evan's蓝灌注心肌,1%氯化三苯基四氮唑(TTC)磷酸缓冲液孵育心肌,CK-MB试剂盒来观察心肌缺血、坏死及肌酸磷酸激酶(CK-MB)和HE染色观察心肌损伤程度,利用蛋白印迹(Western blot)技术探究芍药苷预处理对腺苷酸活化蛋白激酶(AMPKα)的影响。结果芍药苷药物预处理组与缺血再灌注组比较,药物预处理组心梗面积较缺血再灌注组明显减小(P<0.01);CK-MB检测也发现药物预处理组与缺血再灌注组比较能明显降低CK-MB值(P<0.01);HE染色发现药物预处理组较缺血再灌注组能明显减轻大鼠心肌局灶性肿胀、心肌间质水肿,减少炎性细胞浸润及血管充血;蛋白印迹实验发现药物预处理组能明显促进AMPKα蛋白的表达,升高磷酸化AMPKα的表达。结论芍药苷预处理能减轻大鼠在体心肌缺血再灌注损伤,可能通过增加AMPKα蛋白的表达,促进AMPKα磷酸化而实现。

Objective To explore the protective effects of pretreatment with paeoniflorin on the myocardial ischemia reperfusion injury of rats in vivo by activating AMPKα. Methods The SPF grade male SD rats （30 rats） were divided into three groups： sham operation group [the rats＇ chests were opened, but the left anterior descending coronary arteries （LAD） were not ligated], ischemia reperfusion group and paeoniflorin pretreatment group, each with I0 cases. Myocardial ischemia reperfusion injury model was established by cardiac LAD ligation in SD rats. The de- gree of myocardial injury was observed by HE staining and perfusion of 0.5% Evan＇s blue and 1% triphenyl tetrazoli- um chloride （TTC）. Western blot was used to explore the effects of paeoniflorin pretreatment on AMPKα, Results In comparison with ischemic reperfusion group, paeoniflorin pretreatment could decrease infarction area significantly （P〈 0.01） and reduce the value of creatine phosphokinase （CK-MB） significantly （P〈0.01）. HE staining showed paeoniflo- rin pretreatment could significantly relieve myocardial focal swelling, intermyocardial swelling, inflammatory cell in- filtration, and vascular engorgement. Western blot showed that paeoniflorin pretreatment could increase the expression of AMPKa and promote the phosphorylation of AMPKα. Conclusion Pretreatment with paeoniflorin could relieve myocardial ischemia reperfusion injury of rats in vivo. The effect is probably achieved by increasing the expression of AMPKα and promoting phosphorylation ofAMPK α.