“双固一通”电针预处理对心肌缺血再灌注损伤模型大鼠细胞凋亡的机制研究

Study on the Mechanism of "Shuanggu Yitong" Electroacupuncture Preconditioning on Cell Apoptosis in a Rat Model of Myocardial Ischemia Reperfusion Injury

目的 探讨“双固一通”电针法通过调控miR-133a-3p-丝裂原活化蛋白激酶激酶(mitogen-activated protein kinase kinase,MKK)3/MKK 6-p38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinases,p38 MAPK)通路对心肌缺血再灌注(ischemia reperfusion,I/R)损伤(MIRI)模型大鼠的保护作用。方法 将50只大鼠随机分为5组:假手术组(sham group,S组)、模型组(ischemia reperfusion group,I/R组)、电针预处理组(electroacupuncture pretreatment group,EA组)、miR-133-3p抑制剂组(antagomir group,Ant组)、电针预处理+miR-133-3p抑制剂组(EA+Ant组)。采用全自动生化分析仪检测各组大鼠血清学指标,透射电镜观察各组大鼠心肌梗死区组织超微结构,荧光定量PCR检测大鼠心肌梗死组织相关基因表达,免疫组化法检测相关蛋白表达水平。结果与S组比较,I/R组大鼠血清肌酸激酶心肌型同工酶(creatine kinase isoenzymes,CK-MB)和乳酸脱氢酶(lactic dehydrogenase,LDH)水平增加,总抗氧化能力(total antioxidant capacity,T-AOC)水平降低(P<0.05);心肌梗死组织B淋巴细胞瘤(B-cell lymphoma,Bcl)-2相关X蛋白(Bcl-2 associated X protein,Bax)、半胱氨酸天冬氨酸蛋白酶(cystein-asparate protease,Caspase)-3,MKK3,MKK6,p38MAPK表达增加,Bcl-2表达下降(P<0.05)。与I/R组比较,EA组大鼠血清CK-MB和LDH水平降低,T-AOC水平升高(P<0.05);心肌梗死组织Bax、Caspase-3,MKK3,MKK6,p38MAPK表达下降,Bcl-2表达升高(P<0.05)。结论 “双固一通”电针预处理可有效改善心肌缺血再灌注损伤,其机制可能与通过调节miR-133a-3p-MKK3/MKK6-p38MAPK通路抑制细胞凋亡有关。

Objective To explore the protective effect of"Shuanggu Yitong"electroacupuncture on myocardial ischemia-reperfusion(I/R)injury MIRI in rats via regulating the miR-133a-3p-MKK3/MKK6-p38 MAPK(p38 MARK)pathway.Methods 50 rats were randomly divided into 5 groups:sham group(S group),ischemia reperfusion group(I/R group),electroacupuncture pretreatment group(EA group),miR-133-3p antagomir group(Ant group),and electroacupuncture pretreatment+miR-133-3p antagomir group(EA+Ant group).The electrocardiogram of rats in each group was recorded by the BL-420S biological function experiment system.The serological indexes of rats in each group were detected by an automatic biochemical analyzer.The ultrastructure of the myocardial infarction area was observed by transmission electron microscope.The expression of related genes in myocardial infarction tissue was detected by RT qPCR,while the expression level of related proteins was observed by the immunohistochemistry method.Results Compared with the S group,rats in the I/R group had higher level of serum CK-MB and LDH,while lower level of T-AOC(P<0.05),the expression of Bax,Caspase-3 MKK3,MKK6,and p38MAPK were increased,while Bcl-2 was decreased(P<0.05)in the myocardial infarction area of rats in I/R group.Compare with the I/R group,the serum CK-MB and LDH levels of rats decreased,while the T-AOC level increased(P<0.05)in the EA group,the expression of Bax,Caspase-3,MKK3,MKK6,p38MAPK decreased,while Bcl-2 increased(P<0.05)in myocardial infarction tissue of rats in the EA group.Conclusion"Shuanggu Yitong"electroacupuncture pretreatment can effectively ameliorate MIRI via regulating the miR-133a-3p-MKK3/MKK6-p38MAPK pathway to inhibit cell apoptosis.