电针联合运动对肥胖大鼠骨骼肌过氧化物酶体增殖物激活受体γ辅激活因子1α、鸢尾素、腺苷酸活化蛋白激酶的影响
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摘要
目的:观察电针联合运动疗法对高脂饮食诱导的肥胖(DIO)大鼠骨骼肌过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC-1α)、鸢尾素(Irisin)、腺苷酸活化蛋白激酶(AMPK)表达的影响,探讨电针联合运动改善肥胖的机制。方法:SD大鼠随机分为对照组(10只)和高脂组(55只)。采用高脂饮食喂养复制肥胖大鼠模型。高脂组中成功造模的肥胖大鼠随机分为模型组、运动组、电针组和电针联合运动组,每组8只。电针联合运动组和电针组电针"足三里""天枢"穴,每次30min,每周5次,共8周。电针联合运动组和运动组采用跑台训练,16m/min,每天60min,每周5次,共8周。每周测量各组大鼠体质量,采用荧光定量PCR法检测大鼠腓肠肌中PGC-1α、Ⅲ型纤连蛋白组件包含蛋白5(FNDC5)、AMPK mRNA的表达,Western blot法检测大鼠腓肠肌中PGC-1α、FNDC5的表达水平及AMPK的磷酸化水平。结果:模型组大鼠体质量明显高于对照组(P<0.05);PGC-1α、FNDC5mRNA及蛋白表达水平,AMPK mRNA及磷酸化水平明显低于正常组(P<0.05)。电针组、运动组、电针联合运动组体质量明显低于模型组(P<0.05),PGC-1α、FNDC5mRNA及蛋白表达水平,AMPK mRNA及磷酸化水平明显高于模型组(P<0.05)。电针联合运动组体质量明显低于电针组及运动组(P<0.05),PGC-1α、FNDC5mRNA及蛋白表达水平,AMPK mRNA及磷酸化水平明显高于电针组及运动组(P<0.05)。结论:电针和运动均可促进DIO大鼠骨骼肌中PGC-1α、Irisin的表达及AMPK磷酸化水平,且电针联合运动疗效更好,提示电针联合运动疗法可能是通过恢复骨骼肌细胞脂肪酸氧化、改善线粒体功能从而实现减肥的。
Objective To investigate the effect of electroacupuncture(EA)plus treadmill exercise on the expression of peroxisome proliferator activated receptorγcoactivator 1α(PGC-1α),Irisin,AMP-activated protein kinase(AMPK)in skeletal muscle of diet-induced obesity(DIO)rats,so as to explore its mechanism underlying body reduction promotion.Methods Forty-two male SD rats were divided into normal diet(control,n=10),high fat diet(model),EA,treadmill exercise and EA plus treadmill exercise(combination)groups(n=8 in each of the latter 4 groups).The obesity model was established by feeding the rats with high fat diet.EA(2 Hz/15 Hz,1 mA)was applied to bilateral"Zusanli"(ST36)and"Tianshu"(ST25)for 30 min,5 times per week for a total of 8 weeks.Rats of the treadmill exercise group were forced to perform exercise on a treadmill(16 m/min)for 30 min,5 times per week for a total of 8 weeks.Rats in the combination group received the above-mentioned two methods.During the treatment,rats in the control group were fed with normal fodder,rats in other groups were fed with high fat fodder,and their body weight was measured once a week.The expression levels of PGC-1α,fibronectin typeⅢ domain containing 5(FNDC5),AMPK mRNA and protein of skeletal muscle were measured by quantitative real-time PCR and Western blot,respectively.Results After modeling,the body weight was significantly increased(P<0.05),and the expression levels of PGC-1αand FNDC5 mRNA and protein,AMPK mRNA and phosphorylated AMPK(p-AMPK)protein in the skeletal muscle were considerably decreased in the model group relevant to the control group(P<0.05).Following the treatment,the body weight was significantly down-regulated,while the expression levels of PGC-1αand FNDC5 mRNAs and proteins,AMPK mRNA and p-AMPK protein were obviously upregulated in the EA,treadmill exercise and combination groups relevant to the model group(P<0.05).The therapeutic effect of EA plus treadmill exercise was significantly superior to those of both simple EA and simple treadmill exercise in down-regulating the body weight,as well as in up-regulating the expression of PGC-1αand FNDC5 mRNAs and proteins,AMPK mRNA,and p-AMPK protein(P<0.05).Conclusion Both EA and treadmill exercise can significantly increase the expression of PGC-1α,FNDC5 and p-AMPK in skeletal muscle of DIO rats,suggesting their efficacy in restoring fatty acid oxidation in skeletal muscle cells and improving mitochondrial function,which may contribute to their function in body reduction.The therapeutic effect of EA plus treadmill exercise is better than that of simple EA and simple treadmill exercise.
Objective To investigate the effect of electroacupuncture(EA)plus treadmill exercise on the expression of peroxisome proliferator activated receptorγcoactivator 1α(PGC-1α),Irisin,AMP-activated protein kinase(AMPK)in skeletal muscle of diet-induced obesity(DIO)rats,so as to explore its mechanism underlying body reduction promotion.Methods Forty-two male SD rats were divided into normal diet(control,n=10),high fat diet(model),EA,treadmill exercise and EA plus treadmill exercise(combination)groups(n=8 in each of the latter 4 groups).The obesity model was established by feeding the rats with high fat diet.EA(2 Hz/15 Hz,1 mA)was applied to bilateral"Zusanli"(ST36)and"Tianshu"(ST25)for 30 min,5 times per week for a total of 8 weeks.Rats of the treadmill exercise group were forced to perform exercise on a treadmill(16 m/min)for 30 min,5 times per week for a total of 8 weeks.Rats in the combination group received the above-mentioned two methods.During the treatment,rats in the control group were fed with normal fodder,rats in other groups were fed with high fat fodder,and their body weight was measured once a week.The expression levels of PGC-1α,fibronectin typeⅢ domain containing 5(FNDC5),AMPK mRNA and protein of skeletal muscle were measured by quantitative real-time PCR and Western blot,respectively.Results After modeling,the body weight was significantly increased(P<0.05),and the expression levels of PGC-1αand FNDC5 mRNA and protein,AMPK mRNA and phosphorylated AMPK(p-AMPK)protein in the skeletal muscle were considerably decreased in the model group relevant to the control group(P<0.05).Following the treatment,the body weight was significantly down-regulated,while the expression levels of PGC-1αand FNDC5 mRNAs and proteins,AMPK mRNA and p-AMPK protein were obviously upregulated in the EA,treadmill exercise and combination groups relevant to the model group(P<0.05).The therapeutic effect of EA plus treadmill exercise was significantly superior to those of both simple EA and simple treadmill exercise in down-regulating the body weight,as well as in up-regulating the expression of PGC-1αand FNDC5 mRNAs and proteins,AMPK mRNA,and p-AMPK protein(P<0.05).Conclusion Both EA and treadmill exercise can significantly increase the expression of PGC-1α,FNDC5 and p-AMPK in skeletal muscle of DIO rats,suggesting their efficacy in restoring fatty acid oxidation in skeletal muscle cells and improving mitochondrial function,which may contribute to their function in body reduction.The therapeutic effect of EA plus treadmill exercise is better than that of simple EA and simple treadmill exercise.
引文
[1]World Health Organization.Fact Sheet Obesity and Overweight 2013.[WWW document].URL http://www.who.int/mediacentre/factsheets/fs311/en/#(accessed August2014).
[2]CAWLEY J,MEYERHOEFER C.The medical care costs of obesity:an instrumental variables approach[J].J Health Econ,2012,31(1):219-230.
[3]罗妮莎,高梓珊,唐萍萍,等.电针结合行为控制治疗超重及单纯性肥胖实践性随机对照研究的依从性控制策略探讨[J].针刺研究,2017,42(2):183-186.
[4]GARVEY W T,MECHANICK J I,BRETT E M,et al.A-merican association of clinical endocrinologists and American college of endocrinology comprehensive clinical practice guidelines for medical care of patients with obesity[J].Endocr Pract,2016,22(Suppl 3):1-203.
[5]FANG S J,WANG M,ZHENG Y Y,et al.Acupuncture and lifestyle modification treatment for obesity:a meta-analysis[J].Am J Chin Med,2017,45(2):239-254.
[6]HANDSCHIN C,SPIEGELMAN B M.The role of exercise and PGC1αin inflammation and chronic disease[J].Nature,2008,454(7203):463-469.
[7]BOSTRM P,WU J,JEDRYCHOWSKI M P,et al.APGC1-α-dependent myokine that drives brown-fat-like development of white fat and thermogenesis[J].Nature,2012,481(7382):463-468.
[8]CARLING D,VIOLLET B.Beyond energy homeostasis:the expanding role of AMP-activated protein kinase in regulating metabolism[J].Cell Metab,2015,21(6):799-804.
[9]MORINO K,PETERSEN K F,SHULMAN G I.Molecular mechanisms of insulin resistance in humans and their potential links with mitochondrial dysfunction[J].Diabetes,2006,55(Suppl 2):S9-S15.
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[13]王海军,冀来喜.关于大鼠“神阙”穴的定位[J].针刺研究,2007,32(5):312.
[14]BEDFORD T G,TIPTON C M,WILSON N C,et al.Maximum oxygen consumption of rats and its changes with various experimental procedures[J].J Appl Physiol,1979,47(6):1278-1283.
[15]梁翠梅,胡慧,王朝歆,等.针刺治疗腹型肥胖随机对照临床试验[J].针刺研究,2016,41(2):159-162.
[16]陈霞,周仲瑜,黄伟,等.针灸疗法配合饮食运动治疗单纯性肥胖并发高脂血症的临床研究[J].针灸临床杂志,2017,33(7):1-5.
[17]陈霞,黄伟,金熠婷,等.基于复杂网络技术对电针治疗单纯性肥胖处方分析[J].中国针灸,2018,38(3):331-336.
[18]汤月霞,孟建忠,丁亚娟,等.电针对高脂饮食诱导的肥胖小鼠白色脂肪组织中腺苷受体表达的影响[J].针刺研究,2017,42(1):39-44.
[19]LIU Z,SUN F,SU J,et al.Study on action of acupuncture on ventromedial nucleus of hypothalamus in obese rats[J].JTradit Chin Med,2001,21(5):220-224.
[20]刘志诚,孙凤岷,赵东红,等.针刺对单纯性肥胖瘦素和胰岛素抵抗的影响研究[J].中医药学刊,2003,21(1):40-43.
[21]王少锦,徐华洲,肖红玲,等.高频电针刺激对实验性肥胖大鼠脂代谢的调节作用[J].针刺研究,2008,33(3):154-158.
[22]SHEN W X,WANG Y,LU S F,et al.Acupuncture promotes white adipose tissue browning by inducing UCP1expression on DIO mice[J].BMC Complement Altern Med,2014,14:501.
[23]VAUGHAN R A,GANNON N P,BARBERENA M A,et al.Characterization of the metabolic effects of irisin on skeletal muscle in vitro[J].Diabetes Obes Metab,2014,16(8):711-718.
[24]ARITA Y,KIHARA S,OUCHI N,et al.Paradoxical decrease of an adipose-specific protein,adiponectin,in obesity[J].Biochem Biophys Res Commun,1999,257(1):79-83.
[25]于涛,李鹏飞,房国梁,等.大鼠一次性下坡跑运动后FNDC5和Irisin水平变化特点[J].中国运动医学杂志,2016,35(10):908-912.
[2]CAWLEY J,MEYERHOEFER C.The medical care costs of obesity:an instrumental variables approach[J].J Health Econ,2012,31(1):219-230.
[3]罗妮莎,高梓珊,唐萍萍,等.电针结合行为控制治疗超重及单纯性肥胖实践性随机对照研究的依从性控制策略探讨[J].针刺研究,2017,42(2):183-186.
[4]GARVEY W T,MECHANICK J I,BRETT E M,et al.A-merican association of clinical endocrinologists and American college of endocrinology comprehensive clinical practice guidelines for medical care of patients with obesity[J].Endocr Pract,2016,22(Suppl 3):1-203.
[5]FANG S J,WANG M,ZHENG Y Y,et al.Acupuncture and lifestyle modification treatment for obesity:a meta-analysis[J].Am J Chin Med,2017,45(2):239-254.
[6]HANDSCHIN C,SPIEGELMAN B M.The role of exercise and PGC1αin inflammation and chronic disease[J].Nature,2008,454(7203):463-469.
[7]BOSTRM P,WU J,JEDRYCHOWSKI M P,et al.APGC1-α-dependent myokine that drives brown-fat-like development of white fat and thermogenesis[J].Nature,2012,481(7382):463-468.
[8]CARLING D,VIOLLET B.Beyond energy homeostasis:the expanding role of AMP-activated protein kinase in regulating metabolism[J].Cell Metab,2015,21(6):799-804.
[9]MORINO K,PETERSEN K F,SHULMAN G I.Molecular mechanisms of insulin resistance in humans and their potential links with mitochondrial dysfunction[J].Diabetes,2006,55(Suppl 2):S9-S15.
[10]中华人民共和国科学技术部.关于发布《关于善待实验动物的指导性意见》的通知[J].畜牧兽医科技信息,2007,14(4):35-36.
[11]孙志,张中成,刘志诚.营养性肥胖动物模型的实验研究[J].中国药理学通报,2002,18(4):466-467.
[12]华兴邦,周浩良.大鼠穴位图谱的研制[J].实验动物与动物实验,1991,2(1):1-5.
[13]王海军,冀来喜.关于大鼠“神阙”穴的定位[J].针刺研究,2007,32(5):312.
[14]BEDFORD T G,TIPTON C M,WILSON N C,et al.Maximum oxygen consumption of rats and its changes with various experimental procedures[J].J Appl Physiol,1979,47(6):1278-1283.
[15]梁翠梅,胡慧,王朝歆,等.针刺治疗腹型肥胖随机对照临床试验[J].针刺研究,2016,41(2):159-162.
[16]陈霞,周仲瑜,黄伟,等.针灸疗法配合饮食运动治疗单纯性肥胖并发高脂血症的临床研究[J].针灸临床杂志,2017,33(7):1-5.
[17]陈霞,黄伟,金熠婷,等.基于复杂网络技术对电针治疗单纯性肥胖处方分析[J].中国针灸,2018,38(3):331-336.
[18]汤月霞,孟建忠,丁亚娟,等.电针对高脂饮食诱导的肥胖小鼠白色脂肪组织中腺苷受体表达的影响[J].针刺研究,2017,42(1):39-44.
[19]LIU Z,SUN F,SU J,et al.Study on action of acupuncture on ventromedial nucleus of hypothalamus in obese rats[J].JTradit Chin Med,2001,21(5):220-224.
[20]刘志诚,孙凤岷,赵东红,等.针刺对单纯性肥胖瘦素和胰岛素抵抗的影响研究[J].中医药学刊,2003,21(1):40-43.
[21]王少锦,徐华洲,肖红玲,等.高频电针刺激对实验性肥胖大鼠脂代谢的调节作用[J].针刺研究,2008,33(3):154-158.
[22]SHEN W X,WANG Y,LU S F,et al.Acupuncture promotes white adipose tissue browning by inducing UCP1expression on DIO mice[J].BMC Complement Altern Med,2014,14:501.
[23]VAUGHAN R A,GANNON N P,BARBERENA M A,et al.Characterization of the metabolic effects of irisin on skeletal muscle in vitro[J].Diabetes Obes Metab,2014,16(8):711-718.
[24]ARITA Y,KIHARA S,OUCHI N,et al.Paradoxical decrease of an adipose-specific protein,adiponectin,in obesity[J].Biochem Biophys Res Commun,1999,257(1):79-83.
[25]于涛,李鹏飞,房国梁,等.大鼠一次性下坡跑运动后FNDC5和Irisin水平变化特点[J].中国运动医学杂志,2016,35(10):908-912.