电针“百会”“太冲”对自发性高血压大鼠心肌肥大和自噬的影响
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摘要
目的:通过观察电针对自发性高血压大鼠(SHR)心肌细胞超微结构和自噬相关蛋白表达的影响,探究电针调节SHR心肌肥大的内在机制。方法:选用12只12周龄雄性WKY大鼠作为正常组,24只同周龄雄性SHR随机分为模型组和电针组各12只。电针组电针"百会"和右侧"太冲",留针20min,每日1次,共30d。以无创血压仪检测大鼠尾动脉收缩压,超声心动检测大鼠左心室肥厚程度和功能,Masson染色和透射电子显微镜观察心肌组织结构和细胞超微结构的变化,Western blot法检测各组大鼠心肌组织中Beclin-1和LC3表达的变化。结果:模型组大鼠收缩压与正常组比较显著升高(P<0.01),电针组大鼠收缩压与模型组相比显著降低(P<0.01)。与正常组比较,模型组左心室质量指数(LVMI)、左室舒张末期前壁厚度(LVAWd)、左室舒张末期后壁厚度(LVPWd)显著升高(P<0.01),舒张末期左室内径(LVIDd)显著降低(P<0.01);与模型组比较,电针组LVMI、LVAWd和LVPWd明显降低(P<0.01,P<0.05),LVIDd显著升高(P<0.01)。Masson染色和透射电子显微镜结果显示,电针组较模型组心肌组织损伤明显改善,自噬现象明显减少。Western blot结果显示,与正常组比较,模型组心肌组织Beclin-1、LC3-Ⅱ的相对表达量和LC3-Ⅱ/LC3-Ⅰ比值显著升高(P<0.01);与模型组比较,电针组心肌组织Beclin-1、LC3-Ⅱ的相对表达量和LC3-Ⅱ/LC3-Ⅰ比值显著降低(P<0.01,P<0.05)。结论:电针"百会""太冲"可有效改善SHR心肌肥大,其机制可能与电针调节Beclin-1和LC3的表达,从而抑制心肌细胞的过度自噬有关。
Objective To investigate the effect of electroacupuncture(EA)on cardiac hypertrophy and expression of myocardial autophagy-specific proteins in spontaneously hypertensive rats(SHRs).Methods Twelve male Wistar Kyoto(WKY)rats were selected as the normal control group,and 24 male SHRs were randomized into model and EA groups(n=12 in each).EA(2 Hz/15 Hz,1 mA)was applied to"Taichong"(LR3)and"Baihui"(GV20)for 20 min,once a day for successive 30 days.The other two groups only received the same grasping and fixation procedures.The systolic blood pressure(SBP)was measured using a non-invasive blood detector,and the left ventricular function including the left ventricular anterior wall diameter(LVAWd),left ventricular posterior wall diameter(LVPWd)and left ventricular internal diameter at end-diastolic dimension(LVIDd)was detected using a real-time echocardiography imaging system,and the left ventricular mass index(left ventricle weight/body weight,LVMI)was calculated.The expression of Beclin-1 and LC3 proteins in the left ventricle tissue was detected by Western blot.Pathological changes of the myocardial tissue were observed by Masson staining and transmission electron microscope(TEM).Results After the intervention,the elevated SBP was significantly lowered in the EA group relevant to the model group(P<0.01)and the increased LVAWd,LVPWd and LVMI were markedly decreased in the EA group(P<0.01,P<0.05),while the reduced LVIDd was significantly ameliorated in the EA group(P<0.01).Masson staining showed that in SHRs,disorder of arrangement of cardiac myofibers,swelling of cardiomyocytes,widened space among myocardial cells with a large number of collagenous fibers were observed,and under TEM,aggregation and unequal size of myocardial mitochondria with vacuolation and rupture,autophagosomes and autophagic lysosomes were seen.These changes were relatively milder in the EA group.Western blot showed that hypertension induced up-regulation of Beclin-1 and LC3-II and ratio of LC3-II/LC3-I,and down-regulation of LC3-I were significantly suppressed in the EA group(P<0.01,P<0.05).Conclusion EA at LR3 and GV20 may improve the left cardiac function,myocardial morphological changes and the degree of autophagy in SHRs,which is related to its effects in down-regulating the expression of myocardial Beclin-1 and LC3-II and in up-regulating expression of LC3-I.
Objective To investigate the effect of electroacupuncture(EA)on cardiac hypertrophy and expression of myocardial autophagy-specific proteins in spontaneously hypertensive rats(SHRs).Methods Twelve male Wistar Kyoto(WKY)rats were selected as the normal control group,and 24 male SHRs were randomized into model and EA groups(n=12 in each).EA(2 Hz/15 Hz,1 mA)was applied to"Taichong"(LR3)and"Baihui"(GV20)for 20 min,once a day for successive 30 days.The other two groups only received the same grasping and fixation procedures.The systolic blood pressure(SBP)was measured using a non-invasive blood detector,and the left ventricular function including the left ventricular anterior wall diameter(LVAWd),left ventricular posterior wall diameter(LVPWd)and left ventricular internal diameter at end-diastolic dimension(LVIDd)was detected using a real-time echocardiography imaging system,and the left ventricular mass index(left ventricle weight/body weight,LVMI)was calculated.The expression of Beclin-1 and LC3 proteins in the left ventricle tissue was detected by Western blot.Pathological changes of the myocardial tissue were observed by Masson staining and transmission electron microscope(TEM).Results After the intervention,the elevated SBP was significantly lowered in the EA group relevant to the model group(P<0.01)and the increased LVAWd,LVPWd and LVMI were markedly decreased in the EA group(P<0.01,P<0.05),while the reduced LVIDd was significantly ameliorated in the EA group(P<0.01).Masson staining showed that in SHRs,disorder of arrangement of cardiac myofibers,swelling of cardiomyocytes,widened space among myocardial cells with a large number of collagenous fibers were observed,and under TEM,aggregation and unequal size of myocardial mitochondria with vacuolation and rupture,autophagosomes and autophagic lysosomes were seen.These changes were relatively milder in the EA group.Western blot showed that hypertension induced up-regulation of Beclin-1 and LC3-II and ratio of LC3-II/LC3-I,and down-regulation of LC3-I were significantly suppressed in the EA group(P<0.01,P<0.05).Conclusion EA at LR3 and GV20 may improve the left cardiac function,myocardial morphological changes and the degree of autophagy in SHRs,which is related to its effects in down-regulating the expression of myocardial Beclin-1 and LC3-II and in up-regulating expression of LC3-I.
引文
[1]肖庆颖,周春霞,赵淑健,等.心肌肥厚的研究进展[J].河北医科大学学报,2015,36(12):1476-1480.
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[3]辛娟娟,高俊虹,陆凤燕,等.探讨针刺改善高血压心肌肥厚及心功能损害潜在机制的新思路[J].针刺研究,2015,40(4):331-336.
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[5]高玲莉,冯晓东.电针神庭、百会穴对脑缺血再灌注后认知障碍大鼠学习记忆能力及自噬相关基因和蛋白表达的影响[J].康复学报,2016,26(4):17-22.
[6]何坚,黄紫妍,陈伟标,等.电针抑制局灶性脑缺血损伤大鼠皮质细胞自噬的实验研究[J].中国康复医学杂志,2015,30(12):1203-1207.
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[13]李澜,高慧,邢晓雪,等.心肌自噬及其分子机制研究进展[J].中国临床药理学杂志,2015,31(24):2479-2482.
[14] CAO D J,GILLETTE T G,HILL J A.Cardiomyocyte autophagy:remodeling,repairing,and reconstructing the heart[J].Curr Hypertens Rep,2009,11(6):406-411.
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[16] WANG C,WANG Y,MCNUTT M A,et al.Autophagy process is associated with anti-neoplastic function[J].Acta Biochim Biophys Sin(Shanghai),2011,43(6):425-432.
[17]高长久,孟令锴,王晶华.参附汤血中移行成分对异丙肾上腺素所致心肌肥大大鼠心肌自噬及凋亡的影响[J].牡丹江医学院学报,2017,38(2):40-42.
[18]马晓雯,常芸,王世强.长期耐力训练对大鼠心肌细胞自噬相关因子Beclin1和LC3的影响[J].体育科学,2016,36(2):66-71.
[19]杨翼,李章华,柳华,等.穴位电刺激预处理对超负荷训练后大鼠心肌细胞自噬的影响[J].中国运动医学杂志,2012,31(3):253-256.
[20]谭成富,王超,杜琳,等.电针、艾灸预处理对心肌缺血再灌注损伤大鼠心肌自噬相关蛋白LC 3、Beclin 1表达的影响[J].针刺研究,2018,43(1):1-7.
[2] XIONG X,YOU C,FENG Q C,et al.Pulse width modulation electro-acupuncture on cardiovascular remodeling and plasma nitric oxide in spontaneously hypertensive rats[J].Evid Based Complement Alternat Med,2011,2011:812160.
[3]辛娟娟,高俊虹,陆凤燕,等.探讨针刺改善高血压心肌肥厚及心功能损害潜在机制的新思路[J].针刺研究,2015,40(4):331-336.
[4]冯德琳,尹洪娜,费双,等.电针对细胞自噬影响的实验研究进展[J].中国医药导报,2017,14(35):25-28.
[5]高玲莉,冯晓东.电针神庭、百会穴对脑缺血再灌注后认知障碍大鼠学习记忆能力及自噬相关基因和蛋白表达的影响[J].康复学报,2016,26(4):17-22.
[6]何坚,黄紫妍,陈伟标,等.电针抑制局灶性脑缺血损伤大鼠皮质细胞自噬的实验研究[J].中国康复医学杂志,2015,30(12):1203-1207.
[7]王佩.实验针灸学[M].上海:上海科学技术出版社,1994.
[8]熊肇军,董吁钢.心肌肥厚的研究进展[J].中国医学前沿杂志(电子版),2013,5(4):17-22.
[9]赵韶华.Profilin-1在自发性高血压大鼠心肌肥大中的作用及机制研究[D].济南:山东大学,2014.
[10]范金茹,王行宽,熊国强.高血压左室肥厚中医病名、病位、病因病机探讨[J].中医药通报,2005,4(2):19-21.
[11] JIA W,ZHANG Y,SUI M,et al.Effect of acupuncture on the genetic expression of myocardial endothelin-1and atrial natriuretic peptide in rats with stress-induced prehypertension[J].J Trad Chin Med Sci,2017,4(1):82-91.
[12] HUO Z J,LI Q,TIAN G H,et al.The ameliorating effects of long-term electroacupuncture on cardiovascular remodeling in spontaneously hypertensive rats[J].BMC Complement Alterna Med,2014,14:118.
[13]李澜,高慧,邢晓雪,等.心肌自噬及其分子机制研究进展[J].中国临床药理学杂志,2015,31(24):2479-2482.
[14] CAO D J,GILLETTE T G,HILL J A.Cardiomyocyte autophagy:remodeling,repairing,and reconstructing the heart[J].Curr Hypertens Rep,2009,11(6):406-411.
[15]桂亚利,余鹏,张阳阳,等.过度自噬介导心肌肥大心功能不全[J].实用老年医学,2016,30(5):370-372.
[16] WANG C,WANG Y,MCNUTT M A,et al.Autophagy process is associated with anti-neoplastic function[J].Acta Biochim Biophys Sin(Shanghai),2011,43(6):425-432.
[17]高长久,孟令锴,王晶华.参附汤血中移行成分对异丙肾上腺素所致心肌肥大大鼠心肌自噬及凋亡的影响[J].牡丹江医学院学报,2017,38(2):40-42.
[18]马晓雯,常芸,王世强.长期耐力训练对大鼠心肌细胞自噬相关因子Beclin1和LC3的影响[J].体育科学,2016,36(2):66-71.
[19]杨翼,李章华,柳华,等.穴位电刺激预处理对超负荷训练后大鼠心肌细胞自噬的影响[J].中国运动医学杂志,2012,31(3):253-256.
[20]谭成富,王超,杜琳,等.电针、艾灸预处理对心肌缺血再灌注损伤大鼠心肌自噬相关蛋白LC 3、Beclin 1表达的影响[J].针刺研究,2018,43(1):1-7.