胰高血糖素样肽1通过类血管生成因子4调节糖尿病db小鼠棕色脂肪活性的机制

doi:10.3969/j.issn.1006-7795.2020.02.012

摘要/Abstract

摘要： 目的探究胰高血糖素样肽1（glucagon-like peptide 1，GLP-1）通过类血管生成因子4（angiopoietin-like protein 4，ANGPTL4）参与棕色脂肪（brown adipose tissue，BAT）功能的调节。方法小鼠分组后皮下埋泵干预6周，取材前进行葡萄糖耐量试验及pet-CT，取空腹血、棕色脂肪及肝脏进行胆固醇和三酰甘油定量、FPLC、qPCR、Western blotting、转录组测序等实验。结果 GLP-1类似物Exendin-4（Ex-4）改善糖尿病db/db小鼠糖耐量异常，降低外周血和BAT中三酰甘油浓度。Ex-4促进棕色组织UCP-1表达，pet-CT提示活性增加。Ex-4促进BAT中ANGPTL4的表达，ANGPTL4缺失降低了BAT活性。结论 GLP-1促进BAT活性伴有ANGPTL4表达水平的升高，ANGPTL4的缺失降低了BAT活性和功能，可能成为BAT活性调控的治疗靶点。

关键词: 胰高血糖素样肽1, 类血管生成因子4, 棕色脂肪, 活性, 2型糖尿病

Abstract: Objective To explore glucagon-like peptide 1 (GLP-1) whether and how to regulate the function of brown adipose tissue (BAT) though angiopoietin-like protein 4 (ANGPTL4). Methods Mice were randomized and controlled to receive subcutaneous pump intervention for 6 weeks. Before sampling, glucose tolerance test and pet-CT were conducted. Fasting blood, brown fat and liver were taken for cholesterol and triglyceride quantification, FPLC, qPCR, Western blotting, and transcriptometric sequencing, etc. Results GLP-1 analogue exendin-4 (Ex-4) improved abnormal glucose tolerance in diabetic db/db mice and reduced triglyceride levels in peripheral blood and BAT. Ex-4 promoted the expression of UCP-1 in BAT, and pet-CT indicated increased activity; Ex-4 promotes the expression of ANGPTL4 in BAT; the deletion of ANGPTL4 decreased the activity of BAT. Conclusion GLP-1 promoted BAT activity accompanied by increased expression level of ANGPTL4, and the deficiency of ANGPTL4 lowered the activity and function of BAT, which may become a therapeutic target for the regulation of BAT activity.

Key words: glucagon-like peptide 1, angiopoietin-like protein 4, brown adipose tissue, activity, type 2 diabetes mellitus

中图分类号:

R589.2

颜岑, 罗小敏, 冯英梅. 胰高血糖素样肽1通过类血管生成因子4调节糖尿病db小鼠棕色脂肪活性的机制[J]. 首都医科大学学报, 2020, 41(2): 217-224.

Yan Cen, Luo Xiaomin, Feng Yingmei. Mechanism of glucagon-like peptide 1 regulating the activity of brown adipose tissue in diabetic db mice by angiopoietin-like protein 4[J]. Journal of Capital Medical University, 2020, 41(2): 217-224.

参考文献

[1] Weir G,Ramage L E,Akyol M,et al. Substantial metabolic activity of human brown adipose tissue during warm conditions and cold-induced lipolysis of local triglycerides[J]. Cell Metab,2018, 27(6):1348-1355.
[2] Alcala M,Calderon-Dominguez M,Bustos E,et al. Increased inflammation,oxidative stress and mitochondrial respiration in brown adipose tissue from obese mice[J].Sci Rep,2017, 7(1):16082.
[3] Vendrell J,El Bekay R,Peral B,et al. Study of the potential association of adipose tissue glp-1 receptor with obesity and insulin resistance[J].Endocrinology,2011, 152(11):4072-4079.
[4] Janssen A W F,Katiraei S,Bartosinska B,et al. Loss of angiopoietin-like 4(angptl4) in mice with diet-induced obesity uncouples visceral obesity from glucose intolerance partly via the gut microbiota[J]. Diabetologia,2018, 61(6):1447-1458.
[5] Tjeerdema N,Georgiadi A,Jonker J T,et al. Inflammation increases plasma angiopoietin-like protein 4 in patients with the metabolic syndrome and type 2 diabetes[J]. BMJ Open Diabetes Res Care, 2014, 2(1):e000034
[6] Guo X,Gao M,Wang Y,et al. Ldl receptor gene-ablated hamsters:A rodent model of familial hypercholesterolemia with dominant inheritance and diet-induced coronary atherosclerosis[J]. EBioMedicine,2018, 27:214-224.
[7] Wu N N,Zhang C H,Lee H J,et al. Brown adipogenic potential of brown adipocytes and peri-renal adipocytes from human embryo[J]. Sci Rep, 2016, 6:39193.
[8] Pellegrinelli V,Carobbio S,Vidal-Puig A. Adipose tissue plasticity:How fat depots respond differently to pathophysiological cues[J].Diabetologia, 2016, 59(6):1075-1088.
[9] Betz M J,Enerback S. Human brown adipose tissue:What we have learned so far[J]. Diabetes,2015, 64(7):2352-360.
[10] Campbell J E,Drucker D J. Pharmacology,physiology,and mechanisms of incretin hormone action[J].Cell Metab,2013, 17(6):819-837.
[11] Orgaard A,Holst J J. The role of somatostatin in glp-1-induced inhibition of glucagon secretion in mice[J].Diabetologia,2017, 60(9):1731-1739.
[12] Krieger J P,Arnold M,Pettersen K G,et al. Knockdown of glp-1 receptors in vagal afferents affects normal food intake and glycemia[J].Diabetes,2016, 65(1):34-43.
[13] Mangmool S,Hemplueksa P,Parichatikanond W,et al. Epac is required for glp-1r-mediated inhibition of oxidative stress and apoptosis in cardiomyocytes[J]. Mol Endocrinol,2015, 29(4):583-596.
[14] Li J,Zheng J,Wang S,et al. Cardiovascular benefits of native glp-1 and its metabolites:An indicator for glp-1-therapy strategies[J].Front Physiol,2017, 8:15.
[15] Batchuluun B,Inoguchi T,Sonoda N,et al. Metformin and liraglutide ameliorate high glucose-induced oxidative stress via inhibition of pkc-nad (p) h oxidase pathway in human aortic endothelial cells[J].Atherosclerosis,2014, 232(1):156-164.
[16] Muskiet M H A,Tonneijck L,Smits M M,et al. Glp-1 and the kidney:From physiology to pharmacology and outcomes in diabetes[J].Nat Rev Nephrol,2017, 13(10):605-628.
[17] Petit J M,Verges B. Glp-1 receptor agonists in nafld[J].Diabetes Metab,2017, 43(Suppl 1):2S28-2S33.
[18] Khound R,Taher J,Baker C,et al. Glp-1 elicits an intrinsic gut-liver metabolic signal to ameliorate diet-induced vldl overproduction and insulin resistance[J].Arterioscler Thromb Vasc Biol,2017, 37(12):2252-2259.
[19] Heppner K M,Marks S,Holland J,et al. Contribution of brown adipose tissue activity to the control of energy balance by glp-1 receptor signalling in mice[J].Diabetologia,2015, 58(9):2124-2132.
[20] Wan Y,Bao X,Huang J,et al. Novel glp-1 analog supaglutide reduces hfd-induced obesity associated with increased ucp-1 in white adipose tissue in mice[J]. Front Physiol,2017, 8:294.
[21] 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.
[22] Lopez M,Dieguez C,Nogueiras R. Hypothalamic glp-1:The control of bat thermogenesis and browning of white fat[J]. Adipocyte,2015, 4(2):141-145.
[23] Seghieri M,Christensen A S,Andersen A,et al. Future perspectives on glp-1 receptor agonists and glp-1/glucagon receptor co-agonists in the treatment of nafld[J]. Front Endocrinol (Lausanne), 2018, 9:649.
[24] Gupta N A,Mells J,Dunham R M,et al. Glucagon-like peptide-1 receptor is present on human hepatocytes and has a direct role in decreasing hepatic steatosis in vitro by modulating elements of the insulin signaling pathway[J]. Hepatology,2010, 51(5):1584-1592.
[25] Zhu P,Goh Y Y,Chin H F,et al. Angiopoietin-like 4:A decade of research[J]. Biosci Rep,2012, 32(3):211-219.
[26] Clement L C,Avila-Casado C,Mace C,et al. Podocyte-secreted angiopoietin-like-4 mediates proteinuria in glucocorticoid-sensitive nephrotic syndrome[J]. Nat Med,2011, 17(1):117-122.
[27] Ruge T,Sukonina V,Kroupa O,et al. Effects of hyperinsulinemia on lipoprotein lipase,angiopoietin-like protein 4,and glycosylphosphatidylinositol-anchored high-density lipoprotein binding protein 1 in subjects with and without type 2 diabetes mellitus[J].Metabolism, 2012, 61(5):652-660.
[28] Clement L C,Mace C,Avila-Casado C,et al. Circulating angiopoietin-like 4 links proteinuria with hypertriglyceridemia in nephrotic syndrome[J].Nat Med,2014, 20(1):37-46.