朱正威;唐成林;李小宏;杨之雪;杨云昊;庞芳;廖冬梅;郭啸;

• 1:重庆医科大学中医药学院
• 2:重庆医科大学附属第一医院康复科

摘要(Abstract)：

目的:观察电针对衰老性肌萎缩小鼠腓肠肌中促血管生成和蛋白转运相关因子的影响,探讨电针抗衰老性肌萎缩的潜在分子机制。方法:将14只30周龄雄性SAMP8小鼠随机分为模型组和电针组,每组7只,以7只同周龄抗快速衰老SAMR1小鼠为对照组。电针组小鼠予以1 mA、4 Hz的连续波电针干预双侧"足三里""阳陵泉",1次/d,每次20 min,每周6次,干预4周。应用力竭跑台实验检测小鼠运动功能;测量腓肠肌质量并计算与体质量的比值;HE染色法和透射电镜观察腓肠肌形态,并计算腓肠肌横截面积;Western blot法检测腓肠肌中蛋白激酶B(AKT)、磷酸化(p)-AKT、哺乳动物雷帕霉素靶蛋白(mTOR)、p-mTOR、 p70核糖体蛋白S6激酶(p70S6K)、p-p70S6K和缺氧诱导因子-1α(HIF-1α)蛋白的相对表达量;实时荧光定量PCR法检测HIF-1α、血管内皮生长因子A(VEGF-A)、肌肉环指蛋白1(MuRF1)、肌萎缩盒F蛋白(MAFbx)mRNA的相对表达量。结果:与对照组比较,模型组小鼠力竭跑台实验的跑步时间与跑步距离减少(P<0.01),体质量和腓肠肌质量减少(P<0.05,P<0.01),腓肠肌质量/体质量减小(P<0.01),腓肠肌细胞萎缩且横截面积变小(P<0.01),p-AKT、p-mTOR、p-p70S6K和HIF-1α蛋白的相对表达量减少(P<0.01),HIF-1α和VEGF-A mRNA相对表达量减少(P<0.01),MuRF1、MAFbx mRNA相对表达量增加(P<0.01)。与模型组比较,电针干预后小鼠力竭跑台实验的跑步时间与跑步距离增加(P<0.05),腓肠肌质量和腓肠肌质量/体质量增加(P<0.05),腓肠肌萎缩程度减轻且横截面积增大(P<0.01), p-AKT、p-mTOR、p-p70S6K和HIF-1α蛋白的相对表达量增加(P<0.01),HIF-1α和VEGF-A mRNA相对表达量增加(P<0.01),MuRF1、MAFbx mRNA相对表达量减少(P<0.01,P<0.05)。结论:电针可能通过调控腓肠肌促血管生成程序和蛋白转运,延缓小鼠衰老性肌萎缩。

关键词(KeyWords)： 衰老性肌萎缩;电针;缺氧;血管生成;蛋白转运

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金青年基金项目(No.81603689);; 重庆市卫计委科卫联合课题(No.ZY201802113)

作者(Author): 朱正威;唐成林;李小宏;杨之雪;杨云昊;庞芳;廖冬梅;郭啸;

Email:

DOI: 10.13702/j.1000-0607.200081

参考文献(References)：

• [1] 穆光宗,张团.我国人口老龄化的发展趋势及其战略应对[J].华中师范大学学报(人文社会科学版),2011,50(5):29-36.
• [2] ROSENBERG I H.Sarcopenia:origins and clinical relevance[J].J Nutr,1997,127(5 Suppl):990S-991S.
• [3] CHANG Y C,CHEN Y T,LIU H W,et al.Oligonol alle-viates sarcopenia by regulation of signaling pathways involved in protein turnover and mitochondrial quality[J].Mol Nutr Food Res,2019,63(10):e1801102.
• [4] GUO A Y,LEUNG K S,SIU P M,et al.Muscle mass,structural and functional investigations of senescence-accele-rated mouse P8 (SAMP8)[J].Exp Anim,2015,64(4):425-433.
• [5] FAVIER F B,BRITTO F A,FREYSSENET D G,et al.HIF-1-driven skeletal muscle adaptations to chronic hypoxia:molecular insights into muscle physiology[J].Cell Mol Life Sci,2015,72(24):4681-4696.
• [6] NOGUCHI E,OHSAWA H,KOBAYASHI S,et al.The effect of electro-acupuncture stimulation on the muscle blood flow of the hindlimb in anesthetized rats[J].Auton Nerv Syst,1999,75(2-3):78-86.
• [7] WANG R,LUO D,XIAO C,et al.The time course effects of electroacupuncture on promoting skeletal muscle regeneration and inhibiting excessive fibrosis after contusion in rabbits[J].Evid Based Complement Alternat Med,2013,2013:869398.
• [8] 李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:316.
• [9] TAKIGAWA K,MATSUDA R,UCHITOMI R,et al.Effects of long-term physical exercise on skeletal muscles in senescence-accelerated mice (SAMP8)[J].Biosci Biotechnol Biochem,2019,83(3):518-524.
• [10] TADAISHI M,MIURA S,KAI Y,et al.Skeletal muscle-specific expression of PGC-1α-b,an exercise-responsive isoform,increases exercise capacity and peak oxygen uptake[J].PLoS One,2011,6(12):e28290.
• [11] 赵丹丹,唐成林,黄思琴,等.电针干预对大鼠腓肠肌适应性肥大及肌卫星细胞增殖分化的影响[J].针刺研究,2017,42(6):489-495.
• [12] 吴梦佳,唐成林,黄思琴,等.电针对失神经大鼠腓肠肌中叉头蛋白转录因子3A/肌萎缩F-box蛋白及成肌分化抗原的影响[J].针刺研究,2019,44(4):253-257.
• [13] 赵丹丹,唐成林,黄思琴,等.电针干预对失神经肌萎缩大鼠肌卫星细胞分化及肌纤维类型转化的影响[J].针刺研究,2019,44(1):37-42.
• [14] 朱正威,唐成林,李小宏,等.电针通过调控微小RNA对失神经肌萎缩大鼠肌卫星细胞增殖的影响[J].针刺研究,2019,44(9):643-648.
• [15] MIURA S,SAITOH S I,KOKUBUN T,et al.Mitochondrial-targeted antioxidant maintains blood flow,mitochondrial function,and redox balance in old mice following prolonged limb ischemia[J].Int J Mol Sci,2017,18(9):1897.
• [16] SCHEERER N,DEHNE N,STOCKMANN C,et al.Myeloid hypoxia-inducible factor-1α is essential for skeletal muscle regeneration in mice[J].J Immunol,2013,191(1):407-414.
• [17] HIROTA K.Regulation of angiogenesis by hypoxia-inducible factor 1[J].Crit Rev Oncol Hematol,2006,59(1):15-26.
• [18] DI GIULIO C,PETRUCCELLI G,BIANCHI G,et al.Does hypoxia cause sarcopenia?Prevention of hypoxia could reduce sarcopenia[J].J Biol Regul Homeost Agents,2009,23(1):55-58.
• [19] STROKA D M,BURKHARDT T,DESBAILLETS I,et al.HIF-1 is expressed in normoxic tissue and displays an organ-specific regulation under systemic hypoxia[J].FASEB J,2001,15(13):2445-2453.
• [20] PIRKMAJER S,FILIPOVIC D,MARS T,et al.HIF-1alpha response to hypoxia is functionally separated from the glucocorticoid stress response in the in vitro regenerating human skeletal muscle[J].Am J Physiol Regul Integr Comp Physiol,2010,299(6):R1693-R1700.
• [21] NIEMI H,HONKONEN K,KORPISALO P,et al.HIF-1α and HIF-2α induce angiogenesis and improve muscle energy recovery[J].Eur J Clin Invest,2014,44(10):989-999.
• [22] RHOADS R P,JOHNSON R M,RATHBONE C R,et al.Satellite cell-mediated angiogenesis in vitro coincides with a functional hypoxia-inducible factor pathway[J].Am J Physiol Cell Physiol,2009,296(6):C1321-C1328.
• [23] RHOADS R P,FLANN K L,CARDINAL T R,et al.Sa-tellite cells isolated from aged or dystrophic muscle exhibit a reduced capacity to promote angiogenesis in vitro[J].Biochem Biophys Res Commun,2013,440(3):399-404.
• [24] LINDHOLM M E.Skeletal muscle hypoxia-inducible factor-1 and exercise[J].Exp Physiol,2016,101(1):28-32.
• [25] FAVIER F B,COSTES F,DEFOUR A,et al.Downregulation of Akt/mammalian target of rapamycin pathway in skeletal muscle is associated with increased REDD1 expression in response to chronic hypoxia[J].Am J Physiol Regul Integr Comp Physiol,2010,298(6):R1659-R1666.
• [26] SUDHAGAR S,SATHYA S.Rapid non-genomic signalling by 17β-oestradiol through c-Src involves mTOR-dependent expression of HIF-1α in breast cancer cells[J].Br J Cancer,2011,105(7):953-960.
• [27] LIU P,GU Y,LUO J,et al.Inhibition of Src activation reverses pulmonary vascular remodeling in experimental pulmonary arterial hypertension via Akt/mTOR/HIF-1 signaling pathway[J].Exp Cell Res,2019,380(1):36-46.
• [28] 吴梦佳,唐成林,黄思琴,等.电针对大鼠失神经腓肠肌中哺乳动物雷帕霉素靶蛋白/70KD核糖体蛋白S6激酶信号通路的影响[J].中国康复理论与实践,2018,24(9):1022-1026.
• [29] GUMUCIO J P.Atrogin-1,MuRF-1,and sarcopenia[J].Endocrine,2013,43(1):12-21.
• [30] BODINE S C.Skeletal muscle atrophy and the E3 ubiquitin ligases MuRF1 and MAFbx/atrogin-1[J].Am J Physiol Endocrinol Metab,2014,307(6):E469-E484.
• [31] CHAILLOU T,KOULMANN N,MEUNIER A,et al.Ambient hypoxia enhances the loss of muscle mass after extensive injury[J].Pflugers Arch,2014,466(3):587-598.
• [32] LéGER B,DERAVE W,DE BOCK K,et al.Human sarcopenia reveals an increase in SOCS-3 and myostatin and a reduced efficiency of Akt phosphorylation[J].Rejuvenat Res,2008,11(1):163-175B.
• [33] RAUE U,SLIVKA D,JEMIOLO B,et al.Proteolytic gene expression differs at rest and after resistance exercise between young and old women[J].J Gerontol A Biol Sci Med Sci,2007,62(12):1407-1412.
• [34] WHITMAN S A,WACKER M J,RICHMOND S R.Contributions of the ubiquitin-proteasome pathway and apoptosis to human skeletal muscle wasting with age[J].Pflugers Arch,2005,450(6):437-446.