牛蒡子苷对L6骨骼肌细胞中腺苷酸活化蛋白激酶信号通路的影响
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摘要
目的探讨牛蒡子苷在L6骨骼肌细胞中对葡萄糖消耗和腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)信号通路的调节作用。方法用含2%胎牛血清的1640培养基诱导L6骨骼肌细胞分化,直至细胞生长出肌管,分化完成后的L6骨骼肌细胞用含有不同浓度牛蒡子苷的培养基培养24 h后,采用葡萄糖氧化酶法检测细胞葡萄糖消耗,western blot法检测AMPK的亚单位AMPKα和磷酸化腺苷酸活化蛋白激酶(p-AMP-activated protein kinase,p-AMPKα)的蛋白表达水平;以real-time PCR法检测过氧化物酶体增生物激活受体γ共激活因子1α(peroxisome-proliferator-activated receptorγcoactivator-1α,PGC-1α)mRNA表达水平。结果牛蒡子苷浓度为1000 g/L时,可增加L6骨骼肌细胞的葡萄糖消耗,差异显著(P<0.01);与对照组相比,牛蒡子苷呈浓度依赖性的增强L6骨骼肌细胞中p-AMPKα蛋白表达水平;并以相同的趋势增强PGC-1αmRNA表达水平。结论牛蒡子苷增加L6骨骼肌细胞的葡萄糖消耗,并激活AMPK/PGC-1α信号通路,具有潜在的改善胰岛素抵抗的作用。
Objective To investigate the effect of arctiin on glucose consumption and AMPK pathway in L6 myotubes. Methods L6 myoblasts were differentiated to myotubes and treated with or without arctiin. After 24 h,glucose oxidase method was performed at the end of treatment and the AMPKα and p-AMPKα levels were detected by western blot( mammalian AMPK is a trimer structure that composed of alpha,beta,gamma of three subunits,and the content detection of total AMPK and activated AMPK proteins usually refers to the alpha subunit.); The PGC-1α mRNA level were measured with real-time PCR. Results The result of glucose oxidase assay suggested that arctiin increase. Compared with control group,arctiin enhanced the activity of p-AMPKα with a concentration-dependent manner,and arctiin promoted the transcription of PGC-1α with a concentration-dependent manner,too. Conclusion Arctiin increases glucose consumption and activates AMPK / PGC-1α signal pathway in L6 myotubes,which suggests that arctiin could potentially improve insulin resistance.
Objective To investigate the effect of arctiin on glucose consumption and AMPK pathway in L6 myotubes. Methods L6 myoblasts were differentiated to myotubes and treated with or without arctiin. After 24 h,glucose oxidase method was performed at the end of treatment and the AMPKα and p-AMPKα levels were detected by western blot( mammalian AMPK is a trimer structure that composed of alpha,beta,gamma of three subunits,and the content detection of total AMPK and activated AMPK proteins usually refers to the alpha subunit.); The PGC-1α mRNA level were measured with real-time PCR. Results The result of glucose oxidase assay suggested that arctiin increase. Compared with control group,arctiin enhanced the activity of p-AMPKα with a concentration-dependent manner,and arctiin promoted the transcription of PGC-1α with a concentration-dependent manner,too. Conclusion Arctiin increases glucose consumption and activates AMPK / PGC-1α signal pathway in L6 myotubes,which suggests that arctiin could potentially improve insulin resistance.
引文
[1]Association AD.14.Diabetes advocacy[J].Diabetes Care,2015,38(Supplement 1):S86-S87.
[2]Szendroedi J,Phielix E,Roden M.The role of mitochondria in insulin resistance and type 2 diabetes mellitus[J].Nature Reviews Endocrinology,2012,8(2):92-103.
[3]Hansell CAH,Chris S,Ross K,et al.Universal expression and dual function of the atypical chemokine receptor D6 on innate-like B cells in mice[J].Blood,2011,117(20):5413-5424.
[4]Koistinen HA,Galuska D,Chibalin AV,et al.5-amino-imidazole carboxamide riboside increases glucose transport and cell-surface GLUT4 content in skeletal muscle from subjects with type 2 diabetes[J].Diabetes,2003,52(5):1066-1072.
[5]王潞,赵烽,刘珂.牛蒡子苷及牛蒡子苷元的药理作用研究进展[J].中草药,2008,39(3):467-470.
[6]刘冬恋,莫正纪,马松涛,等.牛蒡子苷对糖尿病肾病大鼠肾小球滤过屏障损伤的保护作用[J].华西药学杂志,2011,26(6):536-539.
[7]张淑雅,王小萍,陈昕,等.牛蒡苷抗炎和解热作用研究[J].药物评价研究,2013,36(6):422-425.
[8]Su S,Cheng X,Wink M.Natural lignans from Arctium lappa modulate P-glycoprotein efflux function in multidrug resistant cancer cells[J].Phytomedicine,2015,22(2):301-307.
[9]Lira VA,Benton CR,Yan Z,et al.PGC-1αregulation by exercise training and its influences on muscle function and insulin sensitivity[J].Am J Physiol-Endocr Metab,2010,299(2):E145-E161.
[10]周江,文萍,肖秋金.老年2型糖尿病患者心脏功能改变与胰岛素抵抗的关系[J].东南国防医药,2014,16(3):308-309.
[11]黄艳,赵亚萍,王加林.肥胖相关胰岛素抵抗分子机制的研究进展[J].东南国防医药,2010,12(6):526-529.
[12]Flannery C,Dufour S,Rabol R,et al.Skeletal muscle insulin resistance promotes increased hepatic de novo lipogenesis,hyperlipidemia,and hepatic steatosis in the elderly[J].Diabetes,2012,61(11):2711-2717.
[13]Huang SL,Yu RT,Gong J,et al.Arctigenin,a natural compound,activates AMP-activated protein kinase via inhibition of mitochondria complex I and ameliorates metabolic disorders in ob/ob mice[J].Diabetologia,2012,55(5):1469-1481.
[14]Ruderman NB,Carling D,Prentki M,et al.AMPK,insulin resistance,and the metabolic syndrome[J].J Clin Invest,2013,123(7):2764.
[15]Hardie DG.AMPK:a target for drugs and natural products with effects on both diabetes and cancer[J].Diabetes,2013,62(7):2164-2172.
[16]Buler M,Aatsinki S-M,Izzi V,et al.SIRT5 is under the control of PGC-1αand AMPK and is involved in regulation of mitochondrial energy metabolism[J].FASEB J,2014,28(7):3225-3237.
[17]Ruas JL,White JP,Rao RR,et al.A PGC-1αisoform induced by resistance training regulates skeletal muscle hypertrophy[J].Cell,2012,151(6):1319-1331.
[18]Kristensen JM,Skov V,Petersson SJ,et al.A PGC-1α-and muscle fibre type-related decrease in markers of mitochondrial oxidative metabolism in skeletal muscle of humans with inherited insulin resistance[J].Diabetologia,2014,57(5):1006-1015.
[19]Eckardt K,Taube A,Eckel J.Obesity-associated insulin resistance in skeletal muscle:role of lipid accumulation and physical inactivity[J].Rev Endocr Metab Dis,2011,12(3):163-172.
[2]Szendroedi J,Phielix E,Roden M.The role of mitochondria in insulin resistance and type 2 diabetes mellitus[J].Nature Reviews Endocrinology,2012,8(2):92-103.
[3]Hansell CAH,Chris S,Ross K,et al.Universal expression and dual function of the atypical chemokine receptor D6 on innate-like B cells in mice[J].Blood,2011,117(20):5413-5424.
[4]Koistinen HA,Galuska D,Chibalin AV,et al.5-amino-imidazole carboxamide riboside increases glucose transport and cell-surface GLUT4 content in skeletal muscle from subjects with type 2 diabetes[J].Diabetes,2003,52(5):1066-1072.
[5]王潞,赵烽,刘珂.牛蒡子苷及牛蒡子苷元的药理作用研究进展[J].中草药,2008,39(3):467-470.
[6]刘冬恋,莫正纪,马松涛,等.牛蒡子苷对糖尿病肾病大鼠肾小球滤过屏障损伤的保护作用[J].华西药学杂志,2011,26(6):536-539.
[7]张淑雅,王小萍,陈昕,等.牛蒡苷抗炎和解热作用研究[J].药物评价研究,2013,36(6):422-425.
[8]Su S,Cheng X,Wink M.Natural lignans from Arctium lappa modulate P-glycoprotein efflux function in multidrug resistant cancer cells[J].Phytomedicine,2015,22(2):301-307.
[9]Lira VA,Benton CR,Yan Z,et al.PGC-1αregulation by exercise training and its influences on muscle function and insulin sensitivity[J].Am J Physiol-Endocr Metab,2010,299(2):E145-E161.
[10]周江,文萍,肖秋金.老年2型糖尿病患者心脏功能改变与胰岛素抵抗的关系[J].东南国防医药,2014,16(3):308-309.
[11]黄艳,赵亚萍,王加林.肥胖相关胰岛素抵抗分子机制的研究进展[J].东南国防医药,2010,12(6):526-529.
[12]Flannery C,Dufour S,Rabol R,et al.Skeletal muscle insulin resistance promotes increased hepatic de novo lipogenesis,hyperlipidemia,and hepatic steatosis in the elderly[J].Diabetes,2012,61(11):2711-2717.
[13]Huang SL,Yu RT,Gong J,et al.Arctigenin,a natural compound,activates AMP-activated protein kinase via inhibition of mitochondria complex I and ameliorates metabolic disorders in ob/ob mice[J].Diabetologia,2012,55(5):1469-1481.
[14]Ruderman NB,Carling D,Prentki M,et al.AMPK,insulin resistance,and the metabolic syndrome[J].J Clin Invest,2013,123(7):2764.
[15]Hardie DG.AMPK:a target for drugs and natural products with effects on both diabetes and cancer[J].Diabetes,2013,62(7):2164-2172.
[16]Buler M,Aatsinki S-M,Izzi V,et al.SIRT5 is under the control of PGC-1αand AMPK and is involved in regulation of mitochondrial energy metabolism[J].FASEB J,2014,28(7):3225-3237.
[17]Ruas JL,White JP,Rao RR,et al.A PGC-1αisoform induced by resistance training regulates skeletal muscle hypertrophy[J].Cell,2012,151(6):1319-1331.
[18]Kristensen JM,Skov V,Petersson SJ,et al.A PGC-1α-and muscle fibre type-related decrease in markers of mitochondrial oxidative metabolism in skeletal muscle of humans with inherited insulin resistance[J].Diabetologia,2014,57(5):1006-1015.
[19]Eckardt K,Taube A,Eckel J.Obesity-associated insulin resistance in skeletal muscle:role of lipid accumulation and physical inactivity[J].Rev Endocr Metab Dis,2011,12(3):163-172.