电针“夹脊”穴预处理对心肌缺血再灌注大鼠细胞色素P450信号通路基因表达的影响

Effect of“Jiaji”(EX-B 2)-electroacupuncture Preconditioning on Expression of Cytochrome P 450 Signaling Pathways in Myocardial Ischemia-reperfusion Rats

目的:研究电针"夹脊"穴预处理对心肌缺血再灌注损伤大鼠的心肌保护作用及对心肌细胞色素P 450(CYP 450)信号通路基因表达的影响。方法:Wistar大鼠随机分为假手术组、模型组、夹脊组、内关组、阳陵泉组、曲池组,每组10只。采用结扎左冠状动脉前降支法复制心肌缺血再灌注损伤模型。各电针预处理组造模前针刺相应穴位30min,每天1次,治疗7d。观察各组造模前后ST段位移值,ELISA法检测血清肌酸激酶同工酶(CK-MB)的活性,微板法检测血清乳酸脱氢酶(LDH)活性,RT-PCR法检测心肌CYP 450相关基因cyp 1a1、cyp 2b2、cyp 2c11、cyp 4a2mRNA表达量的变化。结果:心电图ST段位移值变化,结扎30min后,夹脊组、内关组均显著低于模型组(P<0.01);再灌注30min后,内关组与模型组比较显著降低(P<0.01)。与假手术组比较,模型组血清CK-MB及LDH活性均显著升高(P<0.01);与模型组比较,夹脊组、内关组、阳陵泉组血清CK-MB及LDH活性均显著降低(P<0.01);与夹脊组比较,曲池组血清LDH显著升高,曲池组及阳陵泉组血清CK-MB显著升高(P<0.05)。模型组与假手术组比较,心肌cyp 1a 1、cyp 2b2mRNA表达量明显升高(P<0.01),cyp 2c11mRNA表达量降低(P<0.01);夹脊组、内关组、阳陵泉组与模型组比较,cyp 1a1、cyp 4a2mRNA表达量降低(P<0.01),cyp 2b2、cyp 2c11mRNA表达量升高(P<0.01);内关组与夹脊组比较,cyp 1a1mRNA表达量降低,cyp 2b2mRNA表达量升高(P<0.05)。结论:电针"夹脊"穴预处理能够减少心肌缺血再灌注中血清心肌酶的产生,降低心电图ST段,上调心肌CYP 450表氧化酶cyp 2b2、cyp 2c11mRNA表达量,下调ω-羟化酶cyp 1a1、cyp 4a2mRNA表达量,从而起到心肌保护作用。

Objective To investigate the effect of＂Jiaji＂（EX-B 2）-electroacupuncture（EA）preconditioning on the myocardium and expression of cytochrome P 450 signaling pathways of myocardial cells in myocardial ischemia-reperfusion injury（MI/RI）rats.Methods Sixty male Wistar rats were randomly divided into sham operation,model,Jiaji,Neiguan（PC 6）,Yanglingquan（GB 34）and Quchi（LI 11）groups（n=10in each group）.MI/RI model was established by occlusion of the descending anterior branch of the left coronary artery.EA preconditioning was respectively applied to EX-B 2,PC 6,GB 34 and LI 11for30 min,once daily for 7days.The ECG-ST segment was detected before and after the modeling.The contents of serum creatine kinase-MB（CK-MB）and lactate dehydrogenase（LDH）were detected by ELISA and microplate method separately.The expression levels of myocardial CYP 450 related genes cyp 1a1,cyp 2b2,cyp 2c11,cyp 4a2 were detected by Real-time PCR.Results Thirty minutes after ligation the changes of ECG-ST segment displacement of the Jiaji group and Neiguan group were significantly lower than that of the model group（P0.01）,and the change was significantly lower in the Neiguan group than in the model group（P0.01）30minutes after reperfusion.Compared with the sham operation group,the activities of serum CK-MB and LDH were significantly increased in the model group（P0.01）.Compared with the model group,the activities of serum CK-MB and LDH were significantly decreased in the Jiaji,Neiguan and Yanglingquan groups（P0.01）.Compared with the Jiaji group,the activity of serum LDH was significantly increased in the Quchi group,while the activities of serum CK-MB were significantly increased in the Quchi and Yanglingquan groups（P 0.05）.Compared with the sham operation group,the expression levels of myocardial cyp 1a1 and cyp 2b2 mRNAs were significantly higher in the model group（P0.01）,while the expression of cyp 2c11 mRNA was lower（P0.01）.Compared with the model group,the expression levels of cyp 1a1 and cyp 4a2 mRNAs were lower in the Jiaji,Neiguan and Yanglingquan groups（P0.01）,while the expressions of cyp 2b2 and cyp 2c11 mRNAs were higher（P0.01）.Compared with the Jiaji group,the expression of cyp 1a1 mRNA was lower in the Neiguan group,while the expression of cyp 2b2 mRNA was higher（P0.05）.Conclusion EA preconditioning at Jiaji（EX-B 2）can reduce serum myocardial enzymes of myocardial ischemia-reperfusion rats,lower the ST segment of electrocardiogram,up-regulate myocardial CYP 450 epoxygenase cyp 2b2 and cyp 2c11 mRNAs expression,down-regulateω-hydroxylase cyp 1a1 and cyp 4a2 mRNAs expression,thus play a role in myocardial protection.