电针预处理对急性心肌缺血再灌注改进模型大鼠的影响
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摘要
目的:在改进大鼠心肌缺血再灌注模型的基础上,给予电针预处理,观察针刺内关穴对大鼠急性心肌缺血再灌注模型的预防性治疗作用。方法:SD大鼠随机分为正常组、模型组、电针预处理组,选择大鼠双侧"内关"穴进行连续3d的电针预处理后,通过改进的造模方法建立缺血再灌注损伤模型,记录心电变化,并于再灌注结束后,进行伊文思蓝-TTC双染色及心律失常评分,采用多重荧光Western Blot法检测损伤局部心肌组织中Troponin C/Ⅰ蛋白相对表达量及其比值。结果:模型组和电针预处理组大鼠心脏结扎以后ST段显著性上抬,呈典型的"墓碑"样变。电针预处理组自再灌1min后ST段逐渐降低至正常组相应水平,与模型组对应时间点ST段幅值比较均降低(P<0.05)。电针预处理组的危险区比例和心律失常评分较模型组显著减少(P<0.01)。与正常组比较,模型组cTnⅠ相对表达量显著降低(P<0.01),cTnC/Ⅰ的比值显著升高(P<0.01);而电针预处理组的cTnC、cTnⅠ相对表达量及cTnC/Ⅰ的比值均降低。与模型组比较,电针预处理组的cTnC相对表达量及cTnC/Ⅰ的比值显著降低(P<0.01),cTnⅠ相对表达量则显著升高(P<0.01)。结论:改进的急性心肌缺血再灌注模型更加稳定可靠,而电针预处理能够有效的降低心肌缺血再灌注导致的心肌损伤。
Objective: To observe the preventive effect of acupuncture at Neiguan at rats' model of acute myocardial ischemia-reperfusion by being given acupuncture pretreatment on the basis of improving the rat model of myocardial ischemiareperfusion. Methods: SD rats were randomly divided into normal group(N), model group(M), electroacupuncture pretreatment group(A). After three consecutive days of acupuncture preconditioning at Neiguan acupoint on both sides of rats, the model of ischemia-reperfusion injury was established by improved modeling method, and the changes of ECG were recorded. Evans blueTTC double staining and arrhythmia score were taken out after the reperfusion. The relative expression of Troponin C/I protein and its ratio in the injured myocardium were detected by multiple fluorescence Western Blot. Results: The ST segment of rats in group M and group A was significantly elevated after cardiac ligation, showing a typical ‘tombstone' like change. The ST segment of group A gradually decreased to the corresponding level of group N after reperfusion for 1 min, and the amplitude of ST segment at the corresponding time point decreased(P<0.05) compared with that in group M. The risk area proportion and arrhythmia score in group A were significantly reduced compared with group M(P<0.01). Compared with group N, the relative expression of cTnI in group M reduced significantly(P<0.01), and the proportion of cTnC/Ⅰ increased significantly(P<0.01). In group A, the relative expression of cTnC and cTnI and the proportion of cTnC/Ⅰ decreased. Compared with group M, the relative expression of cTnC and the proportion of cTnC/Ⅰ in group A decreased significantly(P<0.01), while the relative expression of cTnI increased significantly(P<0.01). Conclusion: The improved acute myocardial ischemia reperfusion model is more stable and reliable.Electroacupuncture pretreatment can effectively reduce the myocardial injury caused by myocardial ischemia-reperfusion.
Objective: To observe the preventive effect of acupuncture at Neiguan at rats' model of acute myocardial ischemia-reperfusion by being given acupuncture pretreatment on the basis of improving the rat model of myocardial ischemiareperfusion. Methods: SD rats were randomly divided into normal group(N), model group(M), electroacupuncture pretreatment group(A). After three consecutive days of acupuncture preconditioning at Neiguan acupoint on both sides of rats, the model of ischemia-reperfusion injury was established by improved modeling method, and the changes of ECG were recorded. Evans blueTTC double staining and arrhythmia score were taken out after the reperfusion. The relative expression of Troponin C/I protein and its ratio in the injured myocardium were detected by multiple fluorescence Western Blot. Results: The ST segment of rats in group M and group A was significantly elevated after cardiac ligation, showing a typical ‘tombstone' like change. The ST segment of group A gradually decreased to the corresponding level of group N after reperfusion for 1 min, and the amplitude of ST segment at the corresponding time point decreased(P<0.05) compared with that in group M. The risk area proportion and arrhythmia score in group A were significantly reduced compared with group M(P<0.01). Compared with group N, the relative expression of cTnI in group M reduced significantly(P<0.01), and the proportion of cTnC/Ⅰ increased significantly(P<0.01). In group A, the relative expression of cTnC and cTnI and the proportion of cTnC/Ⅰ decreased. Compared with group M, the relative expression of cTnC and the proportion of cTnC/Ⅰ in group A decreased significantly(P<0.01), while the relative expression of cTnI increased significantly(P<0.01). Conclusion: The improved acute myocardial ischemia reperfusion model is more stable and reliable.Electroacupuncture pretreatment can effectively reduce the myocardial injury caused by myocardial ischemia-reperfusion.
引文
[1]国家卫生计生委合理用药专家委员会和中国药师协会.冠心病合理用药指南.中国医学前沿杂志:电子版,2016,8(6):19-108
[2]Wang Q,Liang D,Wang F,et al.Efficacy of electroacupuncture pretreatment for myocardial injury in patients undergoing percutaneous coronary intervention:A randomized clinical trial with a 2-year follow-up.International Journal of Cardiology,2015,194:28-35
[3]Yang L,Yang J,Wang Q,et al.Cardioprotective effects of electroacupuncture pretreatment on patients undergoing heart valve replacement surgery:a randomized controlled trial.Annals of Thoracic Surgery,2010,89(3):781-786
[4]华兴邦,周浩良.大鼠穴位图谱的研制.实验动物与动物实验,1991(1):1-5
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[6]Wong T M, Lee A Y,Tai K K.Effects of drugs interacting with opioid receptors during normal perfusion or ischemia and reperfusion in the isolated rat heart:An attempt to identify cardiac opioid receptor subtype(s)involved in arrhythmogenesis.Journal of Molecular&Cellular Cardiology,1990,22(10):1167-1175
[7]Curtis M J,Walker M J.Quantification of arrhythmias using scoring systems:an examination of seven scores in an in vivo model of regional myocardial ischaemia.Cardiovascular Research,1988,22(9):656-665
[8]李粮辉,陈文华,郑宏.改进的TTC染色法显示大鼠心肌缺血再灌注损伤.中国实验动物学报,2014(5):75-78
[9]Liu H T,Li F,Wang Y M,et al.A comparison of efficacy between conventional and modified methods of the chronic myocardial ischemia/reperfusion model.African Journal of Biotechnology,2009,8(21):5917-5921
[10]高俊虹,付卫星,晋志高,等.电针预治疗保护心肌缺血再灌注损伤——β-肾上腺素受体的耐受机制.针刺研究,2006(1):22-26
[11]邓宇珺,符永恒,谭宁,等.大鼠活体心肌缺血再灌注损伤模型的建立与综合评估.中国病理生理杂志,2011,27(2):412-416
[12]张玮玮,郭永清.大鼠急性心肌缺血再灌注模型的改进.山西医药杂志,2009,38(19):902-903
[13]Mehta P K,Polk D M,Zhang X,et al.A randomized controlled trial of acupuncture in stable ischemic heart disease patients. International Journal of Cardiology,2014,176(2):367-374
[14]Longhurst J C.Electroacupuncture treatment of arrhythmias in myocardial ischemia.American Journal of Physiology Heart&Circulatory Physiology,2007,292(5):H2032
[15]Richter A,Herlitz J,Hjalmarson A.Effect of acupuncture in patients with angina pectoris.European Heart Journal,1991,12(2):175
[16]Gao J,Fu W,Jin Z,et al.A preliminary study on the cardioprotection of acupuncture pretreatment in rats with ischemia and reperfusion:Involvement of cardiac beta-adrenoceptors.Japanese Journal of Physiology,2006,56(4):275
[17]邢冬梅,李彬,朱明军,等.中医药预防急性心肌梗死后心力衰竭的新思路.中华中医药杂志,2017,32(1):196-19
[18]曹杰红,徐宁红,谭华清.丹参酮ⅡA对阿霉素所致大鼠心肌损伤的保护作用.中国医药导报,2014,11(24):25-28
[19]Zhang J,Song J,Xu J,et al.ERK1/2-Egr-1 Signaling pathwaymediated protective effects of electroacupuncture in a mouse model of myocardial ischemia-reperfusion.Evidence-Based Complementray and Alternative Medicine,2014,2014:253075
[20]Palmer S,Kentish J C.The role of troponin C in modulating the Ca2+sensitivity of mammalian skinned cardiac and skeletal muscle fibres.Journal of Physiology,1994,480(1):45
[21]Farah C S,Miyamoto C A,Ramos C H,et al.Structural and regulatory functions of the NH2-and COOH-terminal regions of skeletal muscle troponin I.Journal of Biological Chemistry,1994,269(7):5230-5240
[22]Farah C S,Reinach F C.The troponin complex and regulation of muscle contraction.Faseb Journal,1995,9(9):755-767
[23]Metzger J M,Westfall M V.Covalent and noncovalent modification of thin filamentaction:the essential role of troponin in cardiac muscle regulation.Circulation Research,2004,94:146-158
[24]Putkey J A,Sweeney H L,Campbell S T.Site-directed mutation of the trigger calcium-binding sites in cardiac troponin C.Journal of Biological Chemistry,1989,264(21):12370-12378
[25]Solaro R J,Lee J A,Kentish J C,et al.Effects of acidosis on ventricular muscle from adult and neonatal rats.Circulation Research,1988,63:779-787
[26]Zhou Hao,Wang Jin,Zhu Pingjun,et al.Ripk3 regulates cardiac microvascular reperfusion injury:The role of IP3R-dependent calcium overload,XO-mediated oxidative stress and F-action/filopodia-based cellular migration.Cellular Signalling,2018,45:12-22
[27]高俊虹.电针预处理改善心肌缺血性损伤的β1-肾上腺素能受体信号转导机.北京:中国中医科学院,2006
[28]Day S M,Westfall M V,Fomicheva E V,et al.Histidine button engineered into cardiac troponin I protects the ischemic and failing heart. Nature Medicine,2006,12(2):181-189
[2]Wang Q,Liang D,Wang F,et al.Efficacy of electroacupuncture pretreatment for myocardial injury in patients undergoing percutaneous coronary intervention:A randomized clinical trial with a 2-year follow-up.International Journal of Cardiology,2015,194:28-35
[3]Yang L,Yang J,Wang Q,et al.Cardioprotective effects of electroacupuncture pretreatment on patients undergoing heart valve replacement surgery:a randomized controlled trial.Annals of Thoracic Surgery,2010,89(3):781-786
[4]华兴邦,周浩良.大鼠穴位图谱的研制.实验动物与动物实验,1991(1):1-5
[5]Fenton R A,Galeckas K J,Dobson J G Jr.Endogenous adenosine reduces depression of cardiac function induced by beta-adrenergic stimulation during low flow perfusion.Journal of Molecular&Cellular Cardiology,1995,27(10):2373
[6]Wong T M, Lee A Y,Tai K K.Effects of drugs interacting with opioid receptors during normal perfusion or ischemia and reperfusion in the isolated rat heart:An attempt to identify cardiac opioid receptor subtype(s)involved in arrhythmogenesis.Journal of Molecular&Cellular Cardiology,1990,22(10):1167-1175
[7]Curtis M J,Walker M J.Quantification of arrhythmias using scoring systems:an examination of seven scores in an in vivo model of regional myocardial ischaemia.Cardiovascular Research,1988,22(9):656-665
[8]李粮辉,陈文华,郑宏.改进的TTC染色法显示大鼠心肌缺血再灌注损伤.中国实验动物学报,2014(5):75-78
[9]Liu H T,Li F,Wang Y M,et al.A comparison of efficacy between conventional and modified methods of the chronic myocardial ischemia/reperfusion model.African Journal of Biotechnology,2009,8(21):5917-5921
[10]高俊虹,付卫星,晋志高,等.电针预治疗保护心肌缺血再灌注损伤——β-肾上腺素受体的耐受机制.针刺研究,2006(1):22-26
[11]邓宇珺,符永恒,谭宁,等.大鼠活体心肌缺血再灌注损伤模型的建立与综合评估.中国病理生理杂志,2011,27(2):412-416
[12]张玮玮,郭永清.大鼠急性心肌缺血再灌注模型的改进.山西医药杂志,2009,38(19):902-903
[13]Mehta P K,Polk D M,Zhang X,et al.A randomized controlled trial of acupuncture in stable ischemic heart disease patients. International Journal of Cardiology,2014,176(2):367-374
[14]Longhurst J C.Electroacupuncture treatment of arrhythmias in myocardial ischemia.American Journal of Physiology Heart&Circulatory Physiology,2007,292(5):H2032
[15]Richter A,Herlitz J,Hjalmarson A.Effect of acupuncture in patients with angina pectoris.European Heart Journal,1991,12(2):175
[16]Gao J,Fu W,Jin Z,et al.A preliminary study on the cardioprotection of acupuncture pretreatment in rats with ischemia and reperfusion:Involvement of cardiac beta-adrenoceptors.Japanese Journal of Physiology,2006,56(4):275
[17]邢冬梅,李彬,朱明军,等.中医药预防急性心肌梗死后心力衰竭的新思路.中华中医药杂志,2017,32(1):196-19
[18]曹杰红,徐宁红,谭华清.丹参酮ⅡA对阿霉素所致大鼠心肌损伤的保护作用.中国医药导报,2014,11(24):25-28
[19]Zhang J,Song J,Xu J,et al.ERK1/2-Egr-1 Signaling pathwaymediated protective effects of electroacupuncture in a mouse model of myocardial ischemia-reperfusion.Evidence-Based Complementray and Alternative Medicine,2014,2014:253075
[20]Palmer S,Kentish J C.The role of troponin C in modulating the Ca2+sensitivity of mammalian skinned cardiac and skeletal muscle fibres.Journal of Physiology,1994,480(1):45
[21]Farah C S,Miyamoto C A,Ramos C H,et al.Structural and regulatory functions of the NH2-and COOH-terminal regions of skeletal muscle troponin I.Journal of Biological Chemistry,1994,269(7):5230-5240
[22]Farah C S,Reinach F C.The troponin complex and regulation of muscle contraction.Faseb Journal,1995,9(9):755-767
[23]Metzger J M,Westfall M V.Covalent and noncovalent modification of thin filamentaction:the essential role of troponin in cardiac muscle regulation.Circulation Research,2004,94:146-158
[24]Putkey J A,Sweeney H L,Campbell S T.Site-directed mutation of the trigger calcium-binding sites in cardiac troponin C.Journal of Biological Chemistry,1989,264(21):12370-12378
[25]Solaro R J,Lee J A,Kentish J C,et al.Effects of acidosis on ventricular muscle from adult and neonatal rats.Circulation Research,1988,63:779-787
[26]Zhou Hao,Wang Jin,Zhu Pingjun,et al.Ripk3 regulates cardiac microvascular reperfusion injury:The role of IP3R-dependent calcium overload,XO-mediated oxidative stress and F-action/filopodia-based cellular migration.Cellular Signalling,2018,45:12-22
[27]高俊虹.电针预处理改善心肌缺血性损伤的β1-肾上腺素能受体信号转导机.北京:中国中医科学院,2006
[28]Day S M,Westfall M V,Fomicheva E V,et al.Histidine button engineered into cardiac troponin I protects the ischemic and failing heart. Nature Medicine,2006,12(2):181-189