电针预处理对一次性力竭运动大鼠心肌组织AMPK、PGC-1α的影响
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摘要
目的观察电针预处理对一次性力竭运动大鼠心肌组织5-单磷酸腺苷活化蛋白激酶(AMPK)和过氧化物酶体增殖物激活受体共激活因子-1(PGC-1α)的影响,探讨在一次性力竭运动对心肌损伤状态下,电针预处理对心肌的保护作用机制。方法将30只SD雄性大鼠随机分为空白对照组、一次性力竭运动组及电针预处理组,每组10只。空白组大鼠、一次性力竭运动组大鼠连续10d每天抓握10min,电针预处理组大鼠连续10d取"内关"、"足三里"、"关元"电针干预10min,疏密波,2Hz～100Hz,1mA。第11天一次性力竭运动组及电针预处理组跑台力竭运动后记录力竭运动时间,1h后取材,空白对照组大鼠直接取材。采用比色法检测血清中乳酸脱氢酶(LDH);ELISA检测血清中肌酸激酶同工酶(CK-MB)、中肌钙蛋白T(c TnT)含量;Western Blot测定心肌组织中AMPK、PGC-1α蛋白表达;实时荧光定量PCR测定心肌组织中AMPK m RNA、PGC-1α m RNA基因表达水平。结果电针预处理组大鼠较一次性力竭运动组大鼠力竭运动时间增长(P<0.05);一次性力竭运动组大鼠较空白对照组大鼠血清LDH、CK-MB、cTnT升高(P<0.05),心肌组织AMPK、PGC-1α下降(P<0.05);电针预处理组大鼠较一次性力竭运动组大鼠血清LDH、CK-MB、c TnT下降,心肌组织AMPK、PGC-1α上升(P<0.05)。结论电针预处理可以通过对AMPK、PGC-1α调控,从而能够对一次性力竭运动损伤的心肌起到保护作用。
Objective To observe the effect of electro-acupuncture(EA) pretreatment on 5-adenosine monophosphate activated protein kinase(AMPK) and peroxisome proliferator activated receptor co-activating factor 1(PGC-1α) in myocardium of rats after one-off exhaustive exercise,and to explore the protective mechanism of EA pretreatment on myocardium in the state of myocardial injury caused by one-off exhaustive exercise. Methods Thirty SD male rats were randomly divided intoblank control group,one-off exhaustive exercise group and EA pretreatment group,with 10 rats in each group. The rats in blank control group and one-off exhaustive exercise group grasped for 10 d for 10 min every day. The rats in EA pretreatment group received EA intervention at"Neiguan","Zusanli"and"Guanyuan"for 10 min,density wave,2～100 Hz,1 mA for 10 d. On the11 th day,the time of exhaustive exercise after running on the platform of one-off exhaustive exercise group and EA pretreatment group was recorded,samples were obtained after 1 h,while blank control group were directly obtained. Determination of lactate dehydrogenase(LDH) in serum by colorimetry; the contents of CK-MB and cTnT in serum were detected by ELISA. Western blot was used to determine the expression of AMPK and PGC-1α in myocardial tissue. Real-time fluorescence quantitative PCR was used to determine the expression levels of AMPK m RNA and PGC-1α mRNA in myocardial tissues. Results The time of exhaustive exercise in EA pretreatment group was longer than that in one-off exhaustive exercise group(P<0.05). The serum LDH,CK-MB and cTnT in exhaustive exercise group were higher than those in no treatment control group(P<0.05),while AMPK and PGC-1α in the myocardial tissue were lower(P<0.05). The serum LDH,CK-MB and cTnT in EA preconditioning group decreased,while the myocardial AMPK and PGC-1α increased(P<0.05) than those in one-off exhaustive exercise group.Conclusion EA preconditioning can protect myocardium from one-off exhaustive exercise injury by regulating AMPK and PGC-1α.
Objective To observe the effect of electro-acupuncture(EA) pretreatment on 5-adenosine monophosphate activated protein kinase(AMPK) and peroxisome proliferator activated receptor co-activating factor 1(PGC-1α) in myocardium of rats after one-off exhaustive exercise,and to explore the protective mechanism of EA pretreatment on myocardium in the state of myocardial injury caused by one-off exhaustive exercise. Methods Thirty SD male rats were randomly divided intoblank control group,one-off exhaustive exercise group and EA pretreatment group,with 10 rats in each group. The rats in blank control group and one-off exhaustive exercise group grasped for 10 d for 10 min every day. The rats in EA pretreatment group received EA intervention at"Neiguan","Zusanli"and"Guanyuan"for 10 min,density wave,2～100 Hz,1 mA for 10 d. On the11 th day,the time of exhaustive exercise after running on the platform of one-off exhaustive exercise group and EA pretreatment group was recorded,samples were obtained after 1 h,while blank control group were directly obtained. Determination of lactate dehydrogenase(LDH) in serum by colorimetry; the contents of CK-MB and cTnT in serum were detected by ELISA. Western blot was used to determine the expression of AMPK and PGC-1α in myocardial tissue. Real-time fluorescence quantitative PCR was used to determine the expression levels of AMPK m RNA and PGC-1α mRNA in myocardial tissues. Results The time of exhaustive exercise in EA pretreatment group was longer than that in one-off exhaustive exercise group(P<0.05). The serum LDH,CK-MB and cTnT in exhaustive exercise group were higher than those in no treatment control group(P<0.05),while AMPK and PGC-1α in the myocardial tissue were lower(P<0.05). The serum LDH,CK-MB and cTnT in EA preconditioning group decreased,while the myocardial AMPK and PGC-1α increased(P<0.05) than those in one-off exhaustive exercise group.Conclusion EA preconditioning can protect myocardium from one-off exhaustive exercise injury by regulating AMPK and PGC-1α.
引文
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[15]胡引,顾一煌,张宏如,等.艾灸对力竭运动心肌细胞保护的研究进展[J].针刺研究,2014,39(6):504-507.
[16]Liu SD,Meng WX,Xu L,et al.GAS5 promotes myocardial apoptosis in myocardial ischemia-reperfusion injury via upregulating LAS1 expression[J].Eur Rev Med Pharmacol Sci,2018,22(23):8447-8453.
[17]Palkeeva M,Studneva I,Molokoedov A,et al.Galanin/GalR1-3 system:A promising therapeutic target for myocardial ischemia/reperfusion injury[J].Biomed Pharmacother,2019,109(1):1556-1562.
[18]Koskinas KC,Ndrepepa G,Raber L,et al.Prognostic Impact of Periprocedural Myocardial Infarction in Patients Undergoing Elective Percutaneous Coronary Interventions[J].Circ Cardiovasc Interv,2018,11(12):e006752.
[19]Reznick RM,Shulman GI.The role of AMP-activated protein kinase in mitochondrial biogenesis[J].J Physiol,2006,574(1):33-39.
[20]王艳,黄德强,罗志军.AMPK对线粒体功能的调节[J].中国细胞生物学学报,2013,35(10):1434-1443.
[21]张城林,冯寒,李丽,等.CTRP3经AMPK/PGC-1通路促进心肌细胞线粒体生物生成[J].中国病理生理杂志,2016,32(8):1503.
[22]Jager S,Handschin C,St-Pierre J,et al.AMP-activated protein kinase(AMPK)action in skeletal muscle via direct phos phorylation of PGC-1alpha[J].Proc Natl Acad Sci USA,2007,104(29):12017-12022.
[23]Canto C,Jiang LQ,Deshmukh AS,et al.Interdependence of AMPK and SIRT1 for metabolic adaptation to fasting and exercise in skeletal muscle[J].Cell Metab,2010,11(3):213-219.
[2]胡蔚婧,王华,吴松,等.针刺预处理不同配穴对运动性疲劳大鼠心肌细胞TERT和CGRP表达的影响[J].时珍国医国药,2018,29(7):1771-1773.
[3]胡蔚婧,王华,吴松.针刺预处理不同配穴对延缓运动性疲劳及心脏功能调节的影响[J].湖北中医药大学学报,2018,20(3):5-8.
[4]程津津,任萍萍.学习《关于善待实验动物的指导性意见》后的体会[J].实验动物科学,2011,28(3):78-79.
[5]陈茜,王华,梁凤霞,等.“标本配穴”电针对慢性心肌缺血大鼠心电保护效应及代谢组学的影响[J].针刺研究,2018,43(11):698-704.
[6]丁海丽,谢凤香,杨澎湃,等.递增负荷跑台运动大鼠心肌一氧化氮/一氧化氮合酶的增龄变化[J].中国老年学杂志,2017,37(6):2636-2638.
[7]Sun XJ,Mao JR.Role of Janus kinase 2/signal transducer and activator of transcription 3 signaling pathway in cardioprotection of exercise preconditioning[J].Eur Rev Med Pharmacol Sci,2018,22(15):4975-4986.
[8]吴帆,王华,吴松.针灸预处理防治运动性疲劳的理论探讨[J].湖北中医药大学学报,2018,20(5):51-54.
[9]李虹霖,王玉珏,夏昆鹏,等.不同干预手法作用于足三里穴对运动性疲劳大鼠脑内5-HT的影响[J].针灸临床杂志,2015,31(8):71-72.
[10]华冰.电针内关穴预处理对力竭运动大鼠心肌线粒体NO、游离Ca2+和ATP含量的影响[J].台州学院学报,2012,34(6):44-47.
[11]谭成富,严洁,王超,等.谢文娟针灸预处理对心肌缺血再灌注损伤兔不同时间热休克蛋白27、70、90表达的影响[J].针刺研究,2017,42(1):31-38.
[12]陈晨,孙忠人,吕晓琳,等.针灸预处理改善心肌缺血再灌注损伤机制的研究[J].针灸临床杂志,2016,32(11):102-106.
[13]刘建宇,刘黎明.电针干预对大强度耐力运动机体自由基代谢的影响[J].针刺研究,2013,38(1):48-51.
[14]张皓然,张宏如,卢圣锋,等.逆灸对力竭运动大鼠心肌组织中AMPK、mTOR的影响[J].中国针灸,2017,37(5):521-526.
[15]胡引,顾一煌,张宏如,等.艾灸对力竭运动心肌细胞保护的研究进展[J].针刺研究,2014,39(6):504-507.
[16]Liu SD,Meng WX,Xu L,et al.GAS5 promotes myocardial apoptosis in myocardial ischemia-reperfusion injury via upregulating LAS1 expression[J].Eur Rev Med Pharmacol Sci,2018,22(23):8447-8453.
[17]Palkeeva M,Studneva I,Molokoedov A,et al.Galanin/GalR1-3 system:A promising therapeutic target for myocardial ischemia/reperfusion injury[J].Biomed Pharmacother,2019,109(1):1556-1562.
[18]Koskinas KC,Ndrepepa G,Raber L,et al.Prognostic Impact of Periprocedural Myocardial Infarction in Patients Undergoing Elective Percutaneous Coronary Interventions[J].Circ Cardiovasc Interv,2018,11(12):e006752.
[19]Reznick RM,Shulman GI.The role of AMP-activated protein kinase in mitochondrial biogenesis[J].J Physiol,2006,574(1):33-39.
[20]王艳,黄德强,罗志军.AMPK对线粒体功能的调节[J].中国细胞生物学学报,2013,35(10):1434-1443.
[21]张城林,冯寒,李丽,等.CTRP3经AMPK/PGC-1通路促进心肌细胞线粒体生物生成[J].中国病理生理杂志,2016,32(8):1503.
[22]Jager S,Handschin C,St-Pierre J,et al.AMP-activated protein kinase(AMPK)action in skeletal muscle via direct phos phorylation of PGC-1alpha[J].Proc Natl Acad Sci USA,2007,104(29):12017-12022.
[23]Canto C,Jiang LQ,Deshmukh AS,et al.Interdependence of AMPK and SIRT1 for metabolic adaptation to fasting and exercise in skeletal muscle[J].Cell Metab,2010,11(3):213-219.