电针内关穴对大鼠体外循环后心肌线粒体超微结构及氧化应激的影响  

Effect of electroacupuncture at Neiguan point on mitochondrial ultrastructure and oxidative stress in rats under-going cardiopulmonary bypass

作　　者：涂杰[1] 李涛[1] 梁东科[1] 刘国锋[1] 刘菊梅[1] 韦秋英[1] 张炳东[1] 

机构地区：[1]广西医科大学第一附属医院心血管病研究所手术麻醉室,南宁530021

出　　处：《广东医学》2015年第9期1322-1326,共5页Guangdong Medical Journal

基　　金：广西自然科学基金资助项目(编号:2013GXNSFBA019124)

摘　　要：目的探讨电针内关穴对大鼠体外循环（CPB）后心肌线粒体超微结构和氧化应激反应的影响。方法成年SD大鼠60只,4～6月龄,体重320～420 g,雌雄不限,采用随机数字表法,将大鼠随机分为4组（n=15）：假手术组（S组）、模型组（M组）、内关穴组（EA组）和非穴组（EAN组）。采用尾动脉插管灌注,右颈静脉插管引流建立CPB模型。S组仅行麻醉和动静脉插管;M组行CPB 2 h;EA组和EAN组行CPB 2 h,且在CPB期间实施电针刺激,其中EA组取双侧内关穴,EAN组取双侧内关穴旁开0.5 cm处。麻醉复苏后2 h,处死动物留取左心室心肌组织。透射电镜下观察心肌组织超微结构,行线粒体损伤程度评分;采用黄嘌呤氧化法检测心肌超氧化物歧化酶（SOD）活性,硫代巴比妥酸法检测心肌丙二醛（MDA）含量,高效液相色谱法测定心肌三磷酸腺苷（ATP）含量;采用差速离心法分离线粒体后,用二氯双氢荧光素染色,在激光共聚焦显微镜下检测线粒体活性氧簇（ROS）水平。结果与S组比较,其余3组线粒体损伤评分升高,心肌SOD活性和ATP含量下降,心肌MDA含量和线粒体ROS水平升高,差异有统计学意义（P〈0.05）;与M组比较,EAN组线粒体损伤评分,心肌SOD活性、MDA和ATP含量,线粒体ROS水平差异无统计学意义（P〉0.05）,EA组线粒体损伤评分下降,心肌SOD活性和ATP含量升高,心肌MDA含量和线粒体ROS水平下降,差异有统计学意义（P〈0.05）;与EAN组比较,EA组线粒体损伤评分下降,心肌SOD活性和ATP含量升高,心肌MDA含量和线粒体ROS水平下降,差异有统计学意义（P〈0.05）。结论电针内关穴可减轻大鼠CPB后心肌线粒体超微结构损伤,改善缺血心肌的能量代谢,其机制可能与减轻心肌线粒体氧化应激水平有关。Objective To investigate the effect of electroacupuncture at Neiguan point on mitochondrial ultrastructure and oxidative stress in rats undergoing cardiopulmonary bypass（ CPB）. Methods Sixty adult Sprague- Dawley rats,aged 4 ～ 6 months and weighing 320 ～ 420 g,were randomly divided into 4 groups（ n = 15 each） ： sham operation group（ Group S）,model group（ Group M）,electroacupuncture at Neiguan point group（ Group EA）,and electroacupuncture at no point group（ Group EAN）. CPB model was established by cannulation between tail arteries and right jugular vein. Rats in Group S were anaesthetized and cannulated; rats in Group M were subjected to CPB for 2 h; rats in Group EA and Group EAN were subjected to CPB for 2 h and electroacupunctured during CPB（ electroacupunctured at Neiguan point or at the point 0. 5 cm nearby Neiguan point were given,respectively）. Myocardial specimens from left ventricle 2 h after anesthesia resuscitation were harvested. The myocardial ultrastructures were observed with transmission electron microscope,for assessment of mitochondrial damage score. The contents of malondialdehyde（ MDA） were assayed according to thiobarbituric acid method,the activities of superoxide dismutase（ SOD） were measured by xanthine oxidase method,and the cardiac ATP contents were assayed by high performance liquid chromatography. Mitochondria fractions were isolated by differential centrifugation,and the generation of reactive oxygen species（ ROS） were measured with 2＇,7＇- dichlorodihydrofluorescein diacetate using laser confocal microscope. Results Compared with Group S,significantly higher score of mitochondrial injury,reduced activity of SOD and concentration of ATP,and increased myocardial concentration of MDA and mitochondrial concentration of ROS were observed in the other three groups（ P〈0. 05）. There was no significant difference in the score of mitochondrial injury,myocardial activity of SOD,myocardial concentrations of MDA and ATP,or mitochondrial concen

关 键 词：电针 内关穴 体外循环 线粒体 氧化应激 

分 类 号：R256.302[医药卫生—中医内科学]