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

发布时间：2018-09-05 17:33

【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(P0.05);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(P0.05),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(P0.05);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(P0.05)。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。
[Abstract]:Objective to investigate the effects of electroacupuncture at Neiguan point on myocardial mitochondrial ultrastructure and oxidative stress after cardiopulmonary bypass (CPB) in rats. Methods 60 adult SD rats were randomly divided into 4 groups (n = 15): sham operation group (S group), model group (M group), Neiguan point group (EA group) and non-point group (EAN group). The CPB model was established by caudal artery intubation and right jugular vein drainage. Group S was treated only with anesthesia and arteriovenous intubation. Group M was treated with CPB 2 h EA group and EAN group with CPB 2 h, and with electroacupuncture stimulation during CPB. In the EA group, the bilateral Neiguan acupoints were taken at the side of Neiguan acupoints for 0.5 cm in the EAN group. Two hours after anaesthesia and resuscitation, the animals were sacrificed and left ventricular myocardial tissue was taken. The ultrastructure of myocardial tissue was observed under transmission electron microscope and the degree of mitochondrial damage was evaluated. The activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in myocardium were detected by xanthine oxidation method and thiobarbituric acid method respectively. The content of adenosine triphosphate (ATP) in myocardium was determined by high performance liquid chromatography (HPLC), and the (ROS) level of mitochondria was detected under confocal laser microscope after mitochondria were isolated by differential centrifugation and stained with dichlorodihydrofluorescein. Results compared with group S, the score of mitochondrial damage increased, the activity of SOD and ATP decreased, the content of MDA and ROS of mitochondria increased in the other three groups (P0.05), and compared with the group M, the score of mitochondrial damage in the other three groups was significantly higher than that in group M. There was no significant difference in myocardial SOD activity and ATP content, and there was no significant difference in mitochondrial ROS level (P0.05) in EA group, mitochondrial damage score decreased, myocardial SOD activity and ATP content increased, myocardial MDA content and mitochondrial ROS level decreased. The difference was statistically significant (P0.05); compared with the EAN group, the mitochondrial injury score of EA group decreased, myocardial SOD activity and ATP content increased, myocardial MDA content and mitochondrial ROS level decreased, the difference was statistically significant (P0.05). Conclusion electroacupuncture at Neiguan point can reduce the damage of myocardial mitochondria ultrastructure and improve the energy metabolism of ischemic myocardium after CPB in rats. The mechanism may be related to the reduction of oxidative stress level of myocardial mitochondria.
【作者单位】：广西医科大学第一附属医院心血管病研究所手术麻醉室;
【基金】：广西自然科学基金资助项目(编号:2013GXNSFBA019124)
【分类号】：R245

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