胰升糖素样肽1对肥胖大鼠肝脏组织中Sesn2/AMPK/mTOR信号通路的干预效应
|
摘要
目的:研究胰升糖素样肽1受体激动剂利拉鲁肽对肥胖大鼠肝脏组织Sesn2/AMPK/mTOR信号通路的影响。方法:雄性SD大鼠随机分为正常饮食组(NC组)与高脂饮食组(HF组),喂养12周后,每组取5只评估肥胖大鼠模型建立。HF组再随机分为HF组、低剂量利拉鲁肽组(LG组)、中剂量利拉鲁肽组(MG组)与高剂量利拉鲁肽组(HG组),各组分别给予生理盐水及不同剂量利拉鲁肽(50、100和200μg/kg,皮下注射,每天2次) 4周。16周时测定体重及附睾脂肪指数;取大鼠肝脏组织HE染色观察肝脏脂肪变性情况; Western blot法检测肝脏组织中Sesn2、AMPK、p-AMPK、mTOR和p-mTOR的蛋白水平。结果:HF组大鼠的体重明显高于NC组(P <0. 01)。HF组肝脏细胞出现脂肪变性。与NC组相比,HF组的Sesn2蛋白水平明显降低(P <0. 01),p-AMPK/AMPK水平明显降低(P <0. 01),而p-mTOR/mTOR水平与NC组相比差异没有统计学显著性。利拉鲁肽干预4周后,药物处理组大鼠体重明显低于HF组(P <0. 01),MG与HG组大鼠的附睾脂肪指数较HF组降低(P <0. 01),肝组织脂肪变性较HF组减轻; HG组的Sesn2蛋白水平明显高于HF组(P <0. 01),MG与HG组的p-AMPK/AMPK水平较HF组明显升高(P <0. 01),LG组、MG与HG组p-mTOR/mTOR水平较HF组明显降低(P <0. 01)。结论:胰升糖素样肽1受体激动剂可能通过Sesn2/AMPK/mTOR信号通路影响机体能量代谢,改善肥胖状态。
AIM: To explored the effect of glucagon-like peptide 1 receptor agonist liraglutide on Sesn2/AMPK/mTOR signaling pathway in the liver of obese rats. METHODS: Male SD rats were divided into normal chow( NC) group( n = 12) and high-fat diet( HF) group( n = 33). After 12 weeks,5 rats of each group were used to assess establishment of obese rat model. The rats in HF group were divided into 4 subgroups,HF group,low dose of liraglutide( LG) group,middle dose of liraglutide( MG) group,and high dose of liraglutide( HG) group,and treated with various doses of liraglutide( 0,50,100 and 200 μg/kg) via hypodermic injection twice a day for 4 weeks. The body weight and epididymal fat index of the rats at the 16 th week were measured. The liver tissue fatty degeneration was observed. The protein levels of Sesn2,AMPK,p-AMPK,mTOR and p-mTOR were determined by Western blot. RESULTS: The body weight of rats in HF group was obviously higher than that in NC group( P < 0. 01). Compared with NC group,the levels of Sesn2 and p-AMPK/AMPK were significantly decreased in HF group( P < 0. 01),while the level of p-mTOR/mTOR was not changed. After treatment with liraglutide for 4-week,the body weight of the rats in LG,MG and HG groups was obviously lower than that in HF group( P < 0. 01),and epididymal fat index of the rats in MG and HG groups was obviously lower than that in HF group( P < 0. 01). The protein level of Sesn2 in HG group was obviously higher than that in HF group( P < 0. 01). The level of p-AMPK/AMPK was significantly increased in MG and HG groups( P < 0. 01). The level of p-mTOR/mTOR was significantly increased decreased in LG,MG and HG groups( P < 0. 01). CONCLUSION: Glucagon-like peptide 1 receptor agonist liraglutide affects energy metabolism and improves the state of obesity through Sesn2/AMPK/mTOR signaling pathway.
AIM: To explored the effect of glucagon-like peptide 1 receptor agonist liraglutide on Sesn2/AMPK/mTOR signaling pathway in the liver of obese rats. METHODS: Male SD rats were divided into normal chow( NC) group( n = 12) and high-fat diet( HF) group( n = 33). After 12 weeks,5 rats of each group were used to assess establishment of obese rat model. The rats in HF group were divided into 4 subgroups,HF group,low dose of liraglutide( LG) group,middle dose of liraglutide( MG) group,and high dose of liraglutide( HG) group,and treated with various doses of liraglutide( 0,50,100 and 200 μg/kg) via hypodermic injection twice a day for 4 weeks. The body weight and epididymal fat index of the rats at the 16 th week were measured. The liver tissue fatty degeneration was observed. The protein levels of Sesn2,AMPK,p-AMPK,mTOR and p-mTOR were determined by Western blot. RESULTS: The body weight of rats in HF group was obviously higher than that in NC group( P < 0. 01). Compared with NC group,the levels of Sesn2 and p-AMPK/AMPK were significantly decreased in HF group( P < 0. 01),while the level of p-mTOR/mTOR was not changed. After treatment with liraglutide for 4-week,the body weight of the rats in LG,MG and HG groups was obviously lower than that in HF group( P < 0. 01),and epididymal fat index of the rats in MG and HG groups was obviously lower than that in HF group( P < 0. 01). The protein level of Sesn2 in HG group was obviously higher than that in HF group( P < 0. 01). The level of p-AMPK/AMPK was significantly increased in MG and HG groups( P < 0. 01). The level of p-mTOR/mTOR was significantly increased decreased in LG,MG and HG groups( P < 0. 01). CONCLUSION: Glucagon-like peptide 1 receptor agonist liraglutide affects energy metabolism and improves the state of obesity through Sesn2/AMPK/mTOR signaling pathway.
引文
[1]Lee JH,Budanov AV,Karin M.Sestrins orchestrate cellular metabolism to attenuate aging[J].Cell Metab,2013,18(6):792-801.
[2]Hardie DG,Ross FA,Hawley SA.AMPK:a nutrient and energy sensor that maintains energy homeostasis[J].Nat Rev Mol Cell Biol,2012,13(4):251-262.
[3]Laplante M,Sabatini DM.m TOR signaling in growth control and disease[J].Cell,2012,149(2):274-293.
[4]Park HW,Park H,Ro SH,et al.Hepatoprotective role of Sestrin2 against chronic ER stress[J].Nat Commun,2014,5:4233.
[5]Lee J,Hong SW,Rhee EJ,et al.GLP-1 receptor agonist and non-alcoholic fatty liver disease[J].Diabetes Metab J,2012,36(4):262-267.
[6]王晓晨,敖娜,都健,等.胰升血糖素样肽1对非酒精性脂肪性肝病大鼠氧化应激损伤的干预[J].中华肝脏病杂志,2014,22(10):757-762.
[7]Lee JH,Budanov AV,Talukdar S,et al.Maintenance of metabolic homeostasis by Sestrin2 and Sestrin3[J].Cell Metab,2012,16(3):311-321.
[8]Huynh MK,Kinyua AW,Yang DJ,et al.Hypothalamic AMPK as a regulator of energy homeostasis[J].Neural Plast,2016,2016:2754078.
[9]Kim GT,Lee SH,Kim JI,et al.Quercetin regulates the sestrin 2-AMPK-p38 MAPK signaling pathway and induces apoptosis by increasing the generation of intracellular ROSin a p53-independent manner[J].Int J Mol Med,2014,33(4):863-869.
[10]Copp J,Manning G,Hunter T.TORC-specific phosphorylation of m TOR:phospho-Ser2481 is a marker for intact m TORC2[J].Cancer Res,2009,69(5):1821-1827.
[11]Chang HJ,Ro SH,Cao J,et al.m TOR regulation of autophagy[J].FEBS Lett,2010,584(7):1287-1295.
[12]Rosner M,Siegel N,Valli A,et al.m TOR phosphorylated at S2448 binds to raptor and rictor[J].Amino Acids,2010,38(1):223-228.
[13]Gong H.AMPK/m TOR-mediated inhibition of survivin partly contributes to metformin-induced apoptosis in human gastric cancer cell[J].Cancer Biol Ther,2015,16(1):77-87.
[14]Ropelle ER,Pauli JR,Fernandes MF,et al.A central role for neuronal AMP-activated protein kinase(AMPK)and mammalian target of rapamycin(m TOR)in high-protein diet-induced weight loss[J].Diabetes,2017,66(3):786-787.
[15]Chai D,Wang G,Zhou Z,et al.Insulin increases sestrin2 content by reducing its degradation through the PI3K/m TOR signaling pathway[J].Int J Endocrinol,2015,2015:505849.
[16]Li H,Liu S,Yuan H,et al.Sestrin 2 induces autophagy and attenuates insulin resistance by regulating AMPK signaling in C2C12 myotubes[J].Exp Cell Res,2017,354(1):18-24.
[17]杨庆宇,郜娜.利拉鲁肽通过调节micro RNA-375对胰岛细胞凋亡的影响[J].中国病理生理杂志,2016,32(9):1627-1634.
[18]曾志刚,徐丽姝,关丽嫦,等.利拉鲁肽对非酒精性脂肪肝大鼠脂联素及胰岛素抵抗的影响[J].中国病理生理杂志,2014,30(3):533-537.
[19]王媛妹,张玉超,陈吉翠,等.利拉鲁肽通过激活CAMKK2/AMPK通路促进骨骼肌FNDC5的表达[J].中国病理生理杂志,2017,33(3):475-480.
[20]Lee J,Hong SW,Chae SW,et al.Exendin-4 improves steatohepatitis by increasing Sirt1 expression in high-fat diet-induced obese C57BL/6J mice[J].PLo S One,2012,7(2):e31394.
[21]Salheen SM,Panchapakesan U,Pollock CA,et al.The DPP-4 inhibitor linagliptin and the GLP-1 receptor agonist exendin-4 improve endothelium-dependent relaxation of rat mesenteric arteries in the presence of high glucose[J].Pharmacol Res,2015,94:26-33.
[22]杨晶,敖娜,都健.胰升糖素样肽1受体激动剂对高脂喂养肥胖大鼠胰岛素靶组织中趋化素表达的干预效应[J].中华内分泌代谢杂志,2016,32(11):946-951.
[23]Beiroa D,Imbernon M,Gallego R,et al.GLP-1 agonism stimulates brown adipose tissue thermogenesis and browning through hypothalamic AMPK[J].Diabetes,2014,63(10):3346-3358.
[24]Luo X,Deng L,Lamsal LP,et al.AMP-activated protein kinase alleviates extracellular matrix accumulation in high glucose-induced renal fibroblasts through m TOR signaling pathway[J].Cell Physiol Biochem,2015,35(1):191-200.
[25]He Q,Sha S,Sun L,et al.GLP-1 analogue improves hepatic lipid accumulation by inducing autophagy via AMPK/m TOR pathway[J].Biochem Biophys Res Commun,2016,476(4):196-203.
[2]Hardie DG,Ross FA,Hawley SA.AMPK:a nutrient and energy sensor that maintains energy homeostasis[J].Nat Rev Mol Cell Biol,2012,13(4):251-262.
[3]Laplante M,Sabatini DM.m TOR signaling in growth control and disease[J].Cell,2012,149(2):274-293.
[4]Park HW,Park H,Ro SH,et al.Hepatoprotective role of Sestrin2 against chronic ER stress[J].Nat Commun,2014,5:4233.
[5]Lee J,Hong SW,Rhee EJ,et al.GLP-1 receptor agonist and non-alcoholic fatty liver disease[J].Diabetes Metab J,2012,36(4):262-267.
[6]王晓晨,敖娜,都健,等.胰升血糖素样肽1对非酒精性脂肪性肝病大鼠氧化应激损伤的干预[J].中华肝脏病杂志,2014,22(10):757-762.
[7]Lee JH,Budanov AV,Talukdar S,et al.Maintenance of metabolic homeostasis by Sestrin2 and Sestrin3[J].Cell Metab,2012,16(3):311-321.
[8]Huynh MK,Kinyua AW,Yang DJ,et al.Hypothalamic AMPK as a regulator of energy homeostasis[J].Neural Plast,2016,2016:2754078.
[9]Kim GT,Lee SH,Kim JI,et al.Quercetin regulates the sestrin 2-AMPK-p38 MAPK signaling pathway and induces apoptosis by increasing the generation of intracellular ROSin a p53-independent manner[J].Int J Mol Med,2014,33(4):863-869.
[10]Copp J,Manning G,Hunter T.TORC-specific phosphorylation of m TOR:phospho-Ser2481 is a marker for intact m TORC2[J].Cancer Res,2009,69(5):1821-1827.
[11]Chang HJ,Ro SH,Cao J,et al.m TOR regulation of autophagy[J].FEBS Lett,2010,584(7):1287-1295.
[12]Rosner M,Siegel N,Valli A,et al.m TOR phosphorylated at S2448 binds to raptor and rictor[J].Amino Acids,2010,38(1):223-228.
[13]Gong H.AMPK/m TOR-mediated inhibition of survivin partly contributes to metformin-induced apoptosis in human gastric cancer cell[J].Cancer Biol Ther,2015,16(1):77-87.
[14]Ropelle ER,Pauli JR,Fernandes MF,et al.A central role for neuronal AMP-activated protein kinase(AMPK)and mammalian target of rapamycin(m TOR)in high-protein diet-induced weight loss[J].Diabetes,2017,66(3):786-787.
[15]Chai D,Wang G,Zhou Z,et al.Insulin increases sestrin2 content by reducing its degradation through the PI3K/m TOR signaling pathway[J].Int J Endocrinol,2015,2015:505849.
[16]Li H,Liu S,Yuan H,et al.Sestrin 2 induces autophagy and attenuates insulin resistance by regulating AMPK signaling in C2C12 myotubes[J].Exp Cell Res,2017,354(1):18-24.
[17]杨庆宇,郜娜.利拉鲁肽通过调节micro RNA-375对胰岛细胞凋亡的影响[J].中国病理生理杂志,2016,32(9):1627-1634.
[18]曾志刚,徐丽姝,关丽嫦,等.利拉鲁肽对非酒精性脂肪肝大鼠脂联素及胰岛素抵抗的影响[J].中国病理生理杂志,2014,30(3):533-537.
[19]王媛妹,张玉超,陈吉翠,等.利拉鲁肽通过激活CAMKK2/AMPK通路促进骨骼肌FNDC5的表达[J].中国病理生理杂志,2017,33(3):475-480.
[20]Lee J,Hong SW,Chae SW,et al.Exendin-4 improves steatohepatitis by increasing Sirt1 expression in high-fat diet-induced obese C57BL/6J mice[J].PLo S One,2012,7(2):e31394.
[21]Salheen SM,Panchapakesan U,Pollock CA,et al.The DPP-4 inhibitor linagliptin and the GLP-1 receptor agonist exendin-4 improve endothelium-dependent relaxation of rat mesenteric arteries in the presence of high glucose[J].Pharmacol Res,2015,94:26-33.
[22]杨晶,敖娜,都健.胰升糖素样肽1受体激动剂对高脂喂养肥胖大鼠胰岛素靶组织中趋化素表达的干预效应[J].中华内分泌代谢杂志,2016,32(11):946-951.
[23]Beiroa D,Imbernon M,Gallego R,et al.GLP-1 agonism stimulates brown adipose tissue thermogenesis and browning through hypothalamic AMPK[J].Diabetes,2014,63(10):3346-3358.
[24]Luo X,Deng L,Lamsal LP,et al.AMP-activated protein kinase alleviates extracellular matrix accumulation in high glucose-induced renal fibroblasts through m TOR signaling pathway[J].Cell Physiol Biochem,2015,35(1):191-200.
[25]He Q,Sha S,Sun L,et al.GLP-1 analogue improves hepatic lipid accumulation by inducing autophagy via AMPK/m TOR pathway[J].Biochem Biophys Res Commun,2016,476(4):196-203.