电针对心肌缺血小鼠心肌组织交感神经相关物质的影响
|
摘要
目的:观察电针对心肌缺血模型小鼠心肌组织交感神经相关物质的影响,探讨其作用机制。方法:将30只成年雄性C57BL/6小鼠随机分为假手术组、模型组和电针组,每组10只。模型组和电针组采用冠状动脉左前降支结扎法建立心肌缺血模型。假手术组不进行冠状动脉左前降支结扎,其余步骤同模型组。电针组电针"内关"穴,疏密波,频率2 Hz/100 Hz,电流强度2 mA,每天1次,每次20 min,共治疗5 d;假手术组和模型组仅抓取、固定,不予电针治疗。记录Ⅱ导联心电图并计算△ST值进行模型评价;采用TTC染色评价小鼠心肌梗死面积;免疫组化评价小鼠心肌组织阳性神经纤维密度;Western blot检测小鼠心肌组织神经调节蛋白-1(NRG-1)、酪氨酸羟化酶(TH)、生长相关蛋白-43(GAP-43)的蛋白表达水平。结果:Ⅱ导联心电图显示,与假手术组相比,模型组和电针组S-T段显著抬高(均P<0.01),提示缺血模型成功建立;TTC染色结果显示,与假手术组相比,模型组心肌梗死面积显著增加(P<0.01),与模型组相比,电针组心肌梗死面积显著减小(P<0.01);免疫组化结果显示,与假手术组相比,模型组小鼠心肌TH阳性神经纤维密度显著增加(P<0.01),与模型组相比,电针组心肌组织TH阳性神经纤维密度显著减小(P<0.05);Western blot结果显示,与假手术组相比,模型组TH、NRG-1、GAP-43的蛋白表达水平显著升高(均P<0.01),与模型组相比,电针组心肌组织中TH、GAP-43蛋白表达显著降低(均P<0.01),NRG-1蛋白表达显著升高(P<0.05)。结论:电针可能通过增加心肌缺血小鼠心肌组织中NRG-1蛋白表达水平,降低TH、GAP-43蛋白表达水平,抑制梗死后交感神经过度兴奋,实现心肌保护效应。
Objective To observe the effects of electroacupuncture(EA) on sympathetic nerve-related substance in myocardial tissue in mice with myocardial ischemia(MI), and to explore its possible mechanism. Methods Thirty adult male C57 BL/6 mice were randomly divided into a sham operation group, a model group and an EA group, 10 mice in each one. The model of MI was established in the model group and EA group by ligating the left anterior descending branch of coronary artery. The mice in the sham operation group were not treated with ligating at left anterior descending branch of coronary artery, but the remaining procedure was similar with the model group. The mice in the EA group were treated with EA at "Neiguan"(PC 6) with 2 Hz/100 Hz of frequency and 2 mA of intensity, 20 min per treatment, once a day for totally 5 days. No EA was given for model group and sham operation group. The electrocardiogram was recorded and △ST value was calculated to evaluate the model. TTC staining was applied to evaluate the infarct size. Immunohistochemical(IHC) method was applied to evaluate the positive nerve fiber density in myocardial tissue. Western blot method was applied to test the protein expression levels of neuregulin-1(NRG-1), tyrosine hydroxylase(TH), growth associated protein-43(GAP-43). Results The electrocardiogram(lead II) results indicated compared with the sham operation group,the S-T segments in the model group and EA group were increased obviously(both P<0.01), indicating the MI model was established successfully. The TTC staining results indicated compared with sham operation group, the infarction size was significantly increased in the model group(P<0.01); compared with the model group, the infarction size in the EA group was significantly reduced(P<0.01). The IHC results indicated compared with the sham operation group, the positive nerve fiber density in myocardial was increased in the model group(P<0.01); compared with the model group, the positive nerve fiber density in myocardial was reduced in the EA group(P<0.05). The Western blot results indicated compared with the sham operation group, the expression levels of TH, NRG-1 and GAP-43 were significantly increased in the model group(P<0.01); compared with the model group, the expression level of TH and GAP-43 were significantly reduced(P<0.01) and that of NRG-1 was increased in the EA group(P<0.05). Conclusion EA could increase the expression of NRG-1 and reduce the expression of TH and GAP-43 in myocardial tissues in MI mice, which could suppress sympathetic nerve hyperexcitability after infarction to achieve myocardial protection effect.
Objective To observe the effects of electroacupuncture(EA) on sympathetic nerve-related substance in myocardial tissue in mice with myocardial ischemia(MI), and to explore its possible mechanism. Methods Thirty adult male C57 BL/6 mice were randomly divided into a sham operation group, a model group and an EA group, 10 mice in each one. The model of MI was established in the model group and EA group by ligating the left anterior descending branch of coronary artery. The mice in the sham operation group were not treated with ligating at left anterior descending branch of coronary artery, but the remaining procedure was similar with the model group. The mice in the EA group were treated with EA at "Neiguan"(PC 6) with 2 Hz/100 Hz of frequency and 2 mA of intensity, 20 min per treatment, once a day for totally 5 days. No EA was given for model group and sham operation group. The electrocardiogram was recorded and △ST value was calculated to evaluate the model. TTC staining was applied to evaluate the infarct size. Immunohistochemical(IHC) method was applied to evaluate the positive nerve fiber density in myocardial tissue. Western blot method was applied to test the protein expression levels of neuregulin-1(NRG-1), tyrosine hydroxylase(TH), growth associated protein-43(GAP-43). Results The electrocardiogram(lead II) results indicated compared with the sham operation group,the S-T segments in the model group and EA group were increased obviously(both P<0.01), indicating the MI model was established successfully. The TTC staining results indicated compared with sham operation group, the infarction size was significantly increased in the model group(P<0.01); compared with the model group, the infarction size in the EA group was significantly reduced(P<0.01). The IHC results indicated compared with the sham operation group, the positive nerve fiber density in myocardial was increased in the model group(P<0.01); compared with the model group, the positive nerve fiber density in myocardial was reduced in the EA group(P<0.05). The Western blot results indicated compared with the sham operation group, the expression levels of TH, NRG-1 and GAP-43 were significantly increased in the model group(P<0.01); compared with the model group, the expression level of TH and GAP-43 were significantly reduced(P<0.01) and that of NRG-1 was increased in the EA group(P<0.05). Conclusion EA could increase the expression of NRG-1 and reduce the expression of TH and GAP-43 in myocardial tissues in MI mice, which could suppress sympathetic nerve hyperexcitability after infarction to achieve myocardial protection effect.
引文
[1]Rischpler C.Acute myocardial infarction[J].Q J Nucl Med Mol Imaging,2016,60(3):236-251.
[2]丁亚娟,卢圣锋,陈霞,等.不同时间电针预处理对大鼠心肌缺血损伤及HIF-1α表达的影响[J].南京中医药大学学报,2017,33(1):40-43.
[3]王军蒙,袁璟,蔡云,等.电针对急性心肌缺血模型小鼠心肌组织中炎性反应的影响[J].中国针灸,2018,38(5):513-518.
[4]黄日龙,张伟,东贵荣.电针内关穴对急性心肌缺血家兔心肌组织能量代谢的实验研究[J].针灸临床杂志,2010,26(12):44-48.
[5]D’Elia E,Pascale A,Marchesi N,et al.Novel approaches to the post-myocardial infarction/heart failure neural remodeling[J].Heart Fail Rev,2014,19(5):611-619.
[6]Liu XD,Sun L,Chen JG,et al.Effects of local cardiac denervation on cardiac innervation and ventricular arrhythmia after chronic myocardial infarction[J].PLoS One,2017,12(7):e181322.
[7]侯帅,郑申,王佳宁,等.电针调节自主神经功能作用机制探讨[J].世界最新医学信息文摘,2017,17(76):88-89,92.
[8]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:328.
[9]Cao JM,Fishbein MC,Han JB,et al.Relationship between regional cardiac hyperinnervation and ventricular arrhythmia[J].Circulation,2000,101(16):1960-1969.
[10]Kingma JG,Simard D,Rouleau JR.Influence of cardiac nerve status on cardiovascular regulation and cardioprotection[J].World J Cardiol,2017,9(6):508-520.
[11]胡和生,程文娟,闫素华.心肌梗死后神经重构与室性心律失常[J].中国心血管杂志,2005(6):469-471.
[12]Hu H,Xuan Y,Wang Y,et al.Targeted NGF siRNA delivery attenuates sympathetic nerve sprouting and deteriorates cardiac dysfunction in rats with myocardial infarction[J].PLoS One,2014,9(4):e95106.
[13]Delfiner MS,Siano J,Li Y,et al.Reduced epicardial vagal nerve density and impaired vagal control in a rat myocardial infarction-heart failure model[J].Cardiovasc Pathol,2017,26:21-29.
[14]汪克明,周逸平,周美启,等.电针经脉对急性心肌缺血家兔心交感神经放电活动的影响[J].针刺研究,2005,30(2):106-107.
[15]李明磊,王华,陈泽斌.电针内关穴对AMI大鼠的保护作用及其与心自主神经调节的相关性研究[J].山西中医学院学报,2007,8(3):17-19.
[16]吴子建,蔡荣林,龙迪和,等.电针“神门”“太溪”穴对急性心肌缺血家兔心交感神经电活动的影响[J].针刺研究,2010,35(1):32-36.
[17]Pascal D,Giovannelli A,Gnavi S,et al.Characterization of glial cell models and in vitro manipulation of the neuregulin1/ErbBsystem[J].Biomed Res Int,2014,2014:310215.
[18]Kuramochi Y,Cote G M,Guo X,et al.Cardiac endothelial cells regulate reactive oxygen species-induced cardiomyocyte apoptosis through neuregulin-1beta/erbB4 signaling[J].J Biol Chem,2004,279(49):51141-51147.
[19]Lai X,Zhong L,Fu HX,et al.Effects of neuregulin-1 on autonomic nervous system remodeling post-myocardial infarction in a rat model[J].Neural Regen Res,2017,12(11):1905-1910.
[20]Kobayashi K,Morita S,Sawada H,et al.Targeted disruption of the tyrosine hydroxylase locus results in severe catecholamine depletion and perinatal lethality in mice[J].J Biol Chem,1995,270(45):27235-27243.
[21]张赫楠,李平,白芳芳,等.酪氨酸羟化酶的研究进展[J].国际病理科学与临床杂志,2013,33(5):449-452.
[22]Xu B,Xu H,Cao H,et al.Intermedin improves cardiac function and sympathetic neural remodeling in a rat model of post myocardial infarction heart failure[J].Mol Med Rep,2017,16(2):1723-1730.
[2]丁亚娟,卢圣锋,陈霞,等.不同时间电针预处理对大鼠心肌缺血损伤及HIF-1α表达的影响[J].南京中医药大学学报,2017,33(1):40-43.
[3]王军蒙,袁璟,蔡云,等.电针对急性心肌缺血模型小鼠心肌组织中炎性反应的影响[J].中国针灸,2018,38(5):513-518.
[4]黄日龙,张伟,东贵荣.电针内关穴对急性心肌缺血家兔心肌组织能量代谢的实验研究[J].针灸临床杂志,2010,26(12):44-48.
[5]D’Elia E,Pascale A,Marchesi N,et al.Novel approaches to the post-myocardial infarction/heart failure neural remodeling[J].Heart Fail Rev,2014,19(5):611-619.
[6]Liu XD,Sun L,Chen JG,et al.Effects of local cardiac denervation on cardiac innervation and ventricular arrhythmia after chronic myocardial infarction[J].PLoS One,2017,12(7):e181322.
[7]侯帅,郑申,王佳宁,等.电针调节自主神经功能作用机制探讨[J].世界最新医学信息文摘,2017,17(76):88-89,92.
[8]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:328.
[9]Cao JM,Fishbein MC,Han JB,et al.Relationship between regional cardiac hyperinnervation and ventricular arrhythmia[J].Circulation,2000,101(16):1960-1969.
[10]Kingma JG,Simard D,Rouleau JR.Influence of cardiac nerve status on cardiovascular regulation and cardioprotection[J].World J Cardiol,2017,9(6):508-520.
[11]胡和生,程文娟,闫素华.心肌梗死后神经重构与室性心律失常[J].中国心血管杂志,2005(6):469-471.
[12]Hu H,Xuan Y,Wang Y,et al.Targeted NGF siRNA delivery attenuates sympathetic nerve sprouting and deteriorates cardiac dysfunction in rats with myocardial infarction[J].PLoS One,2014,9(4):e95106.
[13]Delfiner MS,Siano J,Li Y,et al.Reduced epicardial vagal nerve density and impaired vagal control in a rat myocardial infarction-heart failure model[J].Cardiovasc Pathol,2017,26:21-29.
[14]汪克明,周逸平,周美启,等.电针经脉对急性心肌缺血家兔心交感神经放电活动的影响[J].针刺研究,2005,30(2):106-107.
[15]李明磊,王华,陈泽斌.电针内关穴对AMI大鼠的保护作用及其与心自主神经调节的相关性研究[J].山西中医学院学报,2007,8(3):17-19.
[16]吴子建,蔡荣林,龙迪和,等.电针“神门”“太溪”穴对急性心肌缺血家兔心交感神经电活动的影响[J].针刺研究,2010,35(1):32-36.
[17]Pascal D,Giovannelli A,Gnavi S,et al.Characterization of glial cell models and in vitro manipulation of the neuregulin1/ErbBsystem[J].Biomed Res Int,2014,2014:310215.
[18]Kuramochi Y,Cote G M,Guo X,et al.Cardiac endothelial cells regulate reactive oxygen species-induced cardiomyocyte apoptosis through neuregulin-1beta/erbB4 signaling[J].J Biol Chem,2004,279(49):51141-51147.
[19]Lai X,Zhong L,Fu HX,et al.Effects of neuregulin-1 on autonomic nervous system remodeling post-myocardial infarction in a rat model[J].Neural Regen Res,2017,12(11):1905-1910.
[20]Kobayashi K,Morita S,Sawada H,et al.Targeted disruption of the tyrosine hydroxylase locus results in severe catecholamine depletion and perinatal lethality in mice[J].J Biol Chem,1995,270(45):27235-27243.
[21]张赫楠,李平,白芳芳,等.酪氨酸羟化酶的研究进展[J].国际病理科学与临床杂志,2013,33(5):449-452.
[22]Xu B,Xu H,Cao H,et al.Intermedin improves cardiac function and sympathetic neural remodeling in a rat model of post myocardial infarction heart failure[J].Mol Med Rep,2017,16(2):1723-1730.