“标本配穴”电针对慢性心肌缺血模型大鼠线粒体结构及呼吸酶活性的影响

Protective Effect of Primary-secondary point association Electro-acupuncture on Mitochondrial Morphology and Activity of Respiratory Enzymes in Chronic Myocardial Ischemia Model Rats

目的:以心肌线粒体为切入,观察“标本配穴”电针对慢性心肌缺血模型大鼠心肌线粒体结构、GAPDH、CS、CCO的酶活性及心肌组织ATP含量的影响,探讨“标本配穴”电针对慢性心肌缺血模型大鼠心肌线粒体的保护机制。方法:将30只Wistar雄性大鼠随机分为空白组、模型组、标本配穴组,每组10只。模型组、标本配穴组大鼠皮下注射盐酸异丙肾上腺素制备慢性心肌缺血大鼠模型。标本配穴组在造模结束后取“内关”“足三里”“关元”电针干预10 min,疏密波,2~100 Hz,1 mA,连续21天。运用透射电镜观察心肌线粒体结构变化,采用试剂盒检测3-磷酸甘油醛脱氢酶(GAPDH)、柠檬酸合成酶(CS)、细胞色素C氧化酶(CCO)的酶活性及心肌组织ATP含量。结果:与空白组比较,模型组线粒体结构破坏明显,大部分线粒体出现变形空泡化;而与模型组比较,标本配穴组线粒体结构损伤程度明显减轻。与空白组相比,模型组、标本配穴组心肌组织ATP含量、GAPDH、CS、CCO活性均明显降低(P<0.01)。但与模型组相比,标本配穴组4指标均显著增高(P<0.01),且心肌组织ATP含量与GAPDH、CS、CCO活性成线性相关。结论:在慢性心肌缺血状态下,标本配穴电针可提升线粒体呼吸酶活性,增加有氧呼吸产能效率,改善心脏能量代谢,从而减轻心肌细胞的结构破坏,对心肌组织起到保护作用。

Objective:To observe the effects of“primary-secondary point association”electro-acupuncture on the myocardial mitochondria structure,the enzyme activities of GAPDH,CS and CCO and the ATP content of myocardial tissue in rats with chronic myocardial ischemia targeting on the function of mitochondria;and to explore the protective mechanism of“primary-secondary point association”electro-acupuncture on the myocardial mitochondria structure in chronic myocardial ischemia rats.Methods:30 Wistar male rats were randomly divided into the blank group,the model group and the“primary-secondary point association”group,with 10 rats in each group.Rat models of chronic myocardial ischemia were prepared by subcutaneous injection of isoproterenol hydrochloride.The point association group were intervened by electro-needling neiguan(PC6),Zusanli(ST36)and Guanyuan(CV4)for 10mins for 21 consecutive days(density wave,2~100 Hz,1 mA).Transmission electron microscopy was used to observe the structure of the myocardium mitochondrial,and the enzyme activities of 3-phosphoglyceraldehyde dehydrogenase(GAPDH),citrate synthase(CS),cytochrome C oxidase(CCO)and the ATP content of myocardium were detected by the kit.Results:Compared to the blank group,the structure of mitochondria in the model group was destroyed obviously,and most mitochondria were deformed and vacuolated.The degree of mitochondrial structure damage was significantly reduced in the point association group compared to that in the model group.The ATP content and the activities of GAPDH,CS and CCO in myocardial tissue were significantly reduced in the model group and in the point association group(P<0.01);however,the four indicators were significantly higher in the point association group than those in the model group(P<0.01).The ATP content of myocardial tissue was linearly correlated with the activity of GAPDH,CS and CCO.Conclusion:In the condition of chronic myocardial ischemia,the intervention of“primary-secondary point association”electro-acupuncture can improve the activity of respiratory enzymes,increase the productivity of aerobic respiration,improve the cardiac energy metabolism,and reduce the damaged structure of myocardial cells,thus to exert the protective effect on myocardial tissue.