基于核磁共振氢谱技术研究电针“内关”穴对心肌缺血再灌注损伤大鼠血清及心肌组织代谢物的影响  

Effect of Electroacupuncture at “Neiguan” (PC 6)on Serum and Myocardial Metabolites in Rats with Myocardial Ischemia Reperfusion Injury Based on Nuclear Magnetic Resonance Spectroscopy

作　　者：唐雅妮 谭成富 刘薇薇[1] 严洁[1] 王超[1] 刘密[1] 林东海[2] 黄彩华[3] 杜琳[1] 陈美琳[1] 李姣兰 朱鼎铭 

机构地区：[1]湖南中医药大学针灸推拿学院,长沙410007 [2]厦门大学化学化工学院,福建厦门361005 [3]厦门理工学院体育系,福建厦门361005

出　　处：《针刺研究》2018年第3期152-162,共11页Acupuncture Research

基　　金：国家自然科学基金面上项目(No.81574080);湖南省中医药管理局课题(No.201521);湖南省高校创新平台开放基金项目(No.15k093)

摘　　要：目的:基于核磁共振氢谱(1 H NMR)研究电针"内关"穴对心肌缺血再灌注损伤(MIRI)大鼠血清和心肌组织代谢物的影响,探讨电针"内关"穴对MIRI大鼠代谢模式的影响。方法:30只SD雄性大鼠随机分为对照组、模型组、电针组。对照组:不予电针,仅用鼠板束缚大鼠,每次20min,每日1次,持续7d;模型组予心肌缺血再灌注造模:至第7天后结扎冠状动脉左前降支40 min后恢复血流60 min;电针组(频率10Hz/50Hz,脉冲宽度0.5ms,强度1mA):每日电针双侧"内关"穴1次,时间20min,持续7d,于第7天电针后造模。造模结束后,收集3组大鼠的血清和心肌组织。利用1 H NMR进行代谢物检测并利用多元统计方法主成分分析(PCA)、正交偏最小二乘分析(OPLS-DA)进行模式识别。结果:与对照组相比,模型组大鼠血清的亮氨酸、异亮氨酸、缬氨酸、3-羟基丁酸、乳酸、丙氨酸、赖氨酸、醋酸盐、N-乙酰糖蛋白、丙酮、乙酰乙酸、琥珀酸、谷氨酰胺、多不饱和脂肪酸、肌酸、甘油磷酸胆碱、甘氨酸均浓度下降,低密度脂蛋白/极低密度脂蛋白、葡萄糖浓度上升;其中除丙酮、乙酰乙酸和多不饱和脂肪酸外,低密度脂蛋白/极低密度脂蛋白在电针后下降到接近对照组水平,其它代谢物浓度均在电针后上升到接近对照组水平,电针组大鼠血清的代谢模式更接近对照组。与对照组相比,模型组大鼠心肌组织的乳酸、丙氨酸、赖氨酸、谷氨酸、谷氨酰胺、天冬氨酸、甘油磷酸胆碱、肌酸、牛磺酸、甘氨酸、苏氨酸、腺苷一磷酸、烟酰胺腺嘌呤二核苷酸浓度下降,葡萄糖浓度明显上升;其中肌酸、甘油磷酸胆碱、烟酰胺腺嘌呤二核苷酸浓度在电针后上升,葡萄糖浓度在电针后下降;此外,电针组大鼠心肌组织苏氨酸、腺苷一磷酸浓度较模型组进一步下降;电针组大鼠心肌的代谢模式虽然有向对照组偏移的趋势,但与对照组差异还是较大。�Objective We have repeatedly demonstrated that electroacupuncture （EA） of ＂Neiguan＂（PC 6） can im- prove myocardial ischemia in rats. The present study was designed to investigate the metabolomic profile of peripheral blood se- rum and myocardium involving EA-induced improvement of myocardial ischemia-reperfusion injury （MIRI） in rats by using nuclear magnetic resonance spectroscopy. Methods Thirty male SD rats were equally randomized into blank control, model and EA groups. Rats of the control group were only banded for 20 min, once a day for 7 days. The MIRI model was established by occlu- sion of the anterior descending branch of the left coronary artery for 40 rain, followed by reperfusion for 60 rain, and rats of the model group were banded as those in the control group. EA （10 Hz/50 Hz, 1 mA） was applied to bilateral PC 6 for 20 min, once daily for 7 days. The blood samples and left ventricular myocardial tissues were collected for assaying the profiles of differential metabolites using 1H nuclear magnetic resonance （~H NMR） spectroscopy and multivariate statistical analysis such as the principal components analysis （PCA）, partial least squares-discriminant analysis （PLS-DA） and orthogonal PLS-DA （O-PLS-DA） with SIM- CA-P software 12.0. Results A total of 19 differential metabolites （17 down-regulated, 2 up-regulated） in the serum and 14 dif- ferential metabolites （13 down-regulated and 1 up-regulated） in the ischemic left myocardium were identified after MIRI. Of the 19 serum differential metabolites, amino acids （leucine, isoleucine, valine,alanine, lysine, glycine, glutamine）, 3-hydroxy butyric acid （3-HB）, lactic acid, acetate, N-acetyl glycoprotein （NAc）, acetone, acetoacetate, succinate, polyunsaturated fatty acids （PUFA）, creatine, glycerophosphocholine （GPO） were down-regulated; while low density lipoprotein （LDL）, LDL/very low den- sity l ipoprotein（LDL/VLDL） and glucose obviously up-regulated. Of the 14 myocardial differential

关 键 词：电针 心肌缺血再灌注损伤 特征代谢物 血清 左心室肌 代谢组学 核磁共振 

分 类 号：R245.97[医药卫生—针灸推拿学]