基于心脑相关理论探讨电针心经、心包经穴对大脑中动脉梗阻大鼠心、脑细胞凋亡的影响
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摘要
目的:比较电针手厥阴心包经"内关"、手少阴心经"神门"、督脉"水沟"以及足少阴肾经"照海"对大脑中动脉梗阻(MCAO)大鼠的干预效应及对心脑细胞凋亡相关因子的影响,初步探讨电针心包经、心经与心脑的相关性。方法:SD大鼠随机分为内关组、神门组、水沟组、照海组、模型组及正常组,每组8只,采用颈外动脉插入线栓法建立MCAO大鼠模型,各穴位组分别在造模后每12h电针治疗1次,共治疗5次。采用免疫组化染色法检测各组心肌及脑组织细胞凋亡百分比及Bax、Bcl-2表达的变化。结果:与正常组比较,模型组心、脑组织细胞凋亡百分比、促凋亡因子Bax的表达均增多(P<0.01)。与模型组比较,内关组、神门组心、脑细胞凋亡百分比均不同程度减少(P<0.01,P<0.05),水沟组脑细胞凋亡百分比减少(P<0.05);内关组、神门组、水沟组脑组织Bax的表达减少(P<0.05),Bcl-2的表达增高(P<0.01,P<0.05);内关组心肌Bax的表达减少(P<0.05),内关组、神门组心肌Bcl-2表达增多(P<0.01,P<0.05)。与照海组比较,内关组心、脑细胞凋亡指数、Bax表达均减少(P<0.01,P<0.05),神门组心肌细胞凋亡的指数减少(P<0.05);内关组、水沟组脑组织Bcl-2表达高于照海组(P<0.05),内关组、神门组心肌Bcl-2表达高于照海组(P<0.01,P<0.05)。结论:脑缺血损伤继发心肌损害提示脑病及心,脑心二者之间具有密切的病理相关性。而针刺心包经、心经可较好地抑制脑心细胞凋亡。
Objective To compare the effect of electroacupuncture(EA)of"Neiguan"(PC6)of the Pericardium Meridian,"Shenmen"(HT7)of the Heart Meridian,"Shuigou"(GV26)of the Governor Vessel and"Zhaohai"(KI6)of the Kidney Meridian on myocardial and cerebral cell apoptosis in cerebral ischemia(CI)rats,so as to explore its mechanism underlying improvement of CI based on the theory of"Heart-brain Correlation".Methods Forty-eight SD rats(half male and half female)were randomly divided into normal control,model,PC6,HT7,GV26 and KI6 groups(n=8 in each one).The CI model was established by occlusion of the middle cerebral artery(MCAO).EA(4 Hz/20 Hz,1.5 mA)was applied to the right PC6,HT7,GV26 or KI6 respectively for 30 min,once every 12 hfor 5 times.The cell apoptosis of the ischemic myocardial and cerebral tissues was detected by TUNEL method,and the expression of cerebral and myocardial Bax and Bcl-2 was determined by using immunohistochemistry.Results Following modeling,the cell apoptosis percentages and Bax-positive cells of both myocardial and cerebral tissues were significantly increased in the model group in comparison with the control group(P<0.01).After EA intervention,the cerebral apoptotic percentage and cerebral Bax-positive cells in the cerebral tissue of the PC6,HT7 and GV26 groups,and the myocardial apoptosis percentage in the PC6 and HT7 groups,as well as the myocardial Bax-positive cells in the PC6 group were obviously decreased(P<0.05,P<0.01),while the cerebral Bcl-2 positive cells in the PC6,HT7 and GV26 groups,and the myocardial Bcl-2 positive cells in the PC6 and HT7 groups were significantly increased relevant to the model group(P<0.01,P<0.05).No significant changes were found in the KI 6 group in the cell apoptosis index and percentages and Baxand Bcl-2-positive cells of both myocardium and cerebral cortex tissues compared with the model group(P>0.05).Conclusion EA stimulation of PC6 and HT7 can inhibit CI injury induced cell apoptosis of cerebral and myocardial tissues in CI rats,which is possibly associated with its effects in down-regulating Bax expression and up-regulating Bcl-2 expression of both myocardial and cerebral tissues.
Objective To compare the effect of electroacupuncture(EA)of"Neiguan"(PC6)of the Pericardium Meridian,"Shenmen"(HT7)of the Heart Meridian,"Shuigou"(GV26)of the Governor Vessel and"Zhaohai"(KI6)of the Kidney Meridian on myocardial and cerebral cell apoptosis in cerebral ischemia(CI)rats,so as to explore its mechanism underlying improvement of CI based on the theory of"Heart-brain Correlation".Methods Forty-eight SD rats(half male and half female)were randomly divided into normal control,model,PC6,HT7,GV26 and KI6 groups(n=8 in each one).The CI model was established by occlusion of the middle cerebral artery(MCAO).EA(4 Hz/20 Hz,1.5 mA)was applied to the right PC6,HT7,GV26 or KI6 respectively for 30 min,once every 12 hfor 5 times.The cell apoptosis of the ischemic myocardial and cerebral tissues was detected by TUNEL method,and the expression of cerebral and myocardial Bax and Bcl-2 was determined by using immunohistochemistry.Results Following modeling,the cell apoptosis percentages and Bax-positive cells of both myocardial and cerebral tissues were significantly increased in the model group in comparison with the control group(P<0.01).After EA intervention,the cerebral apoptotic percentage and cerebral Bax-positive cells in the cerebral tissue of the PC6,HT7 and GV26 groups,and the myocardial apoptosis percentage in the PC6 and HT7 groups,as well as the myocardial Bax-positive cells in the PC6 group were obviously decreased(P<0.05,P<0.01),while the cerebral Bcl-2 positive cells in the PC6,HT7 and GV26 groups,and the myocardial Bcl-2 positive cells in the PC6 and HT7 groups were significantly increased relevant to the model group(P<0.01,P<0.05).No significant changes were found in the KI 6 group in the cell apoptosis index and percentages and Baxand Bcl-2-positive cells of both myocardium and cerebral cortex tissues compared with the model group(P>0.05).Conclusion EA stimulation of PC6 and HT7 can inhibit CI injury induced cell apoptosis of cerebral and myocardial tissues in CI rats,which is possibly associated with its effects in down-regulating Bax expression and up-regulating Bcl-2 expression of both myocardial and cerebral tissues.
引文
[1]栾坤,倪光夏.脑梗死后线粒体介导的细胞凋亡通路及针刺干预作用的研究进展[J].中医药现代化,2015,17(12):2627-2630.
[2]陈小娱,张秋玲,白波.电针对脑缺血再灌注大鼠脑组织细胞线粒体膜电位及凋亡的影响[J].针刺研究,2008,33(2):107-110.
[3]李荣,郭景春,程介士.督脉穴位电针对暂时性脑缺血所致神经细胞死亡的影响[J].针刺研究,2003,28(1):10-16.
[4]卜渊,耿德勤,曾因明.针刺调节脑缺血再灌后细胞凋亡相关基因表达的研究进展[J].针刺研究,2003,28(4):288-291.
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[6]邹晓静,施静,刘敬,等.电针对脑缺血再灌注大鼠大脑皮层去甲肾上腺素及细胞凋亡的影响[J].针刺研究,2005,30(4):203-207.
[7]邓宇琚,谭宁,曾红科,等.脑利钠肽预处理对在体大鼠缺血再灌注心肌细胞凋亡及bcl-2、Bax表达的影响[J].2010,90(48):3431-3434.
[8] LONGA E Z,WEINSTEIN P R,CARLSON S,et al.Reversible middle cerebral artery occlusion without craniectomy in rat[J].Stroke,1989,20(1):84.
[9] XIE Z,KOYAMA T,SUZUKI J,et al.Coronary reperfusion following ischemia:different expression of bcl-2and bax proteins,and cardiomyocyte apoptosis[J].Jpn Heart J,2001,42(6):759-770.
[10]李常法,赵驻军,李亚,等.针刺组穴对脑缺血大鼠脑细胞凋亡相关蛋白表达研究[J].针灸临床杂志,2006,22(7):57-58.
[11] FU Y C,CHI C S,YIN S C,et a1.Norepinephrine induces apoptosis in neonatal rat endothelial ceils via down-regulation of Bcl2and activation of h-adrenergic and caspase-2pathways[J].Cardiovasc Res,2004,61(1):143.
[12]冉群芳,倪光夏.针刺对脑缺血后细胞凋亡相关基因表达影响的研究进展[J].上海针灸杂志,2007,26(7):46-48.
[13]孙世晓,刘泓雨,杨春壮,等.电针对局灶性脑缺血大鼠半暗带神经细胞凋亡及凋亡相关基因Bcl-2、Bax、c-Fos蛋白表达的影响[J].中医药学报,2011,39(1):65-68.
[14] YAMAMURA T,OTANI H,NAKAO Y,et al.IGF-1differentially regulates bcl-xl and bax and confers myocardial protection in the rat heart[J].Am J Physiol Heart Circ Physiol,2001,280(3):H 1191-1200.
[15]张俭欣,金龙云,赵德信.电针对大鼠脑缺血再灌注损伤后神经元细胞凋亡影响及机制的实验研究[J].中国实验诊断学,2006,10(6):673-674.
[16]刘英,郭世杰,孙景辉,等.缺氧缺血性脑损伤新生大鼠心肌细胞凋亡及凋亡蛋白Bax、Bcl-2表达的变化[J].临床儿科杂志,2009,27(12):1172-1176.
[17]朱永磊,宋小鸽.针刺对细胞凋亡的影响[J].中医药临床杂志,2007,19(2):102-104.
[18]周美启,周逸平.心主二经论[J].中国针灸,2004,24(4):272-274.
[19]刘保岩,心脑相关性的中西医理论探讨[J].现代中医药,2012,32(6):48-50.
[20]白芳芳,李平,马洪皓,等.浅述心脑相关理论[J].世界中西医结合杂志,2011,11(1):29-31.
[21]关梓桐,徐雅.试述中医心、脑、神志相关性的研究进展[J].世界中医药,2014,9(9):1243-1246.
[22]赵涛,伍海勤.深化脑心同治研究提高临床诊疗效果[J].中国中西医结合杂志,2013,33(12):1593-1595.
[23]关风光,黄丽钗.浅析基于心脑相关理论舒解脑卒中患者的压力[J].福建中医药,2011,42(1):63-64.
[2]陈小娱,张秋玲,白波.电针对脑缺血再灌注大鼠脑组织细胞线粒体膜电位及凋亡的影响[J].针刺研究,2008,33(2):107-110.
[3]李荣,郭景春,程介士.督脉穴位电针对暂时性脑缺血所致神经细胞死亡的影响[J].针刺研究,2003,28(1):10-16.
[4]卜渊,耿德勤,曾因明.针刺调节脑缺血再灌后细胞凋亡相关基因表达的研究进展[J].针刺研究,2003,28(4):288-291.
[5]余晓慧,孙国杰.针刺对局灶性脑缺血大鼠脑细胞凋亡及Bcl-2蛋白表达的影响[J].针刺研究,2004,29(1):15-17.
[6]邹晓静,施静,刘敬,等.电针对脑缺血再灌注大鼠大脑皮层去甲肾上腺素及细胞凋亡的影响[J].针刺研究,2005,30(4):203-207.
[7]邓宇琚,谭宁,曾红科,等.脑利钠肽预处理对在体大鼠缺血再灌注心肌细胞凋亡及bcl-2、Bax表达的影响[J].2010,90(48):3431-3434.
[8] LONGA E Z,WEINSTEIN P R,CARLSON S,et al.Reversible middle cerebral artery occlusion without craniectomy in rat[J].Stroke,1989,20(1):84.
[9] XIE Z,KOYAMA T,SUZUKI J,et al.Coronary reperfusion following ischemia:different expression of bcl-2and bax proteins,and cardiomyocyte apoptosis[J].Jpn Heart J,2001,42(6):759-770.
[10]李常法,赵驻军,李亚,等.针刺组穴对脑缺血大鼠脑细胞凋亡相关蛋白表达研究[J].针灸临床杂志,2006,22(7):57-58.
[11] FU Y C,CHI C S,YIN S C,et a1.Norepinephrine induces apoptosis in neonatal rat endothelial ceils via down-regulation of Bcl2and activation of h-adrenergic and caspase-2pathways[J].Cardiovasc Res,2004,61(1):143.
[12]冉群芳,倪光夏.针刺对脑缺血后细胞凋亡相关基因表达影响的研究进展[J].上海针灸杂志,2007,26(7):46-48.
[13]孙世晓,刘泓雨,杨春壮,等.电针对局灶性脑缺血大鼠半暗带神经细胞凋亡及凋亡相关基因Bcl-2、Bax、c-Fos蛋白表达的影响[J].中医药学报,2011,39(1):65-68.
[14] YAMAMURA T,OTANI H,NAKAO Y,et al.IGF-1differentially regulates bcl-xl and bax and confers myocardial protection in the rat heart[J].Am J Physiol Heart Circ Physiol,2001,280(3):H 1191-1200.
[15]张俭欣,金龙云,赵德信.电针对大鼠脑缺血再灌注损伤后神经元细胞凋亡影响及机制的实验研究[J].中国实验诊断学,2006,10(6):673-674.
[16]刘英,郭世杰,孙景辉,等.缺氧缺血性脑损伤新生大鼠心肌细胞凋亡及凋亡蛋白Bax、Bcl-2表达的变化[J].临床儿科杂志,2009,27(12):1172-1176.
[17]朱永磊,宋小鸽.针刺对细胞凋亡的影响[J].中医药临床杂志,2007,19(2):102-104.
[18]周美启,周逸平.心主二经论[J].中国针灸,2004,24(4):272-274.
[19]刘保岩,心脑相关性的中西医理论探讨[J].现代中医药,2012,32(6):48-50.
[20]白芳芳,李平,马洪皓,等.浅述心脑相关理论[J].世界中西医结合杂志,2011,11(1):29-31.
[21]关梓桐,徐雅.试述中医心、脑、神志相关性的研究进展[J].世界中医药,2014,9(9):1243-1246.
[22]赵涛,伍海勤.深化脑心同治研究提高临床诊疗效果[J].中国中西医结合杂志,2013,33(12):1593-1595.
[23]关风光,黄丽钗.浅析基于心脑相关理论舒解脑卒中患者的压力[J].福建中医药,2011,42(1):63-64.