药学学报, 2016, 51(6): 919-925
引用本文:
周莹, 李秀丽, 易琴, 张晓琳, 田金英, 王冬梅, 吴松, 叶菲. 化合物WS090152治疗小鼠单纯性脂肪肝的实验研究[J]. 药学学报, 2016, 51(6): 919-925.
ZHOU Ying, LI Xiu-li, YI Qin, ZHANG Xiao-lin, TIAN Jin-ying, WANG Dong-mei, WU Song, YE Fei. Investigating the effects of compound WS090152 on non-alcoholic fatty liver in mice[J]. Acta Pharmaceutica Sinica, 2016, 51(6): 919-925.

化合物WS090152治疗小鼠单纯性脂肪肝的实验研究
周莹1, 李秀丽2, 易琴1, 张晓琳1, 田金英1, 王冬梅1, 吴松1, 叶菲1
1. 中国医学科学院、北京协和医学院药物研究所, 新药作用机制研究与药效评价北京市重点实验室, 天然药物活性物质与功能国家重点实验室, 北京 100050;
2. 赤峰学院医学院, 内蒙古 赤峰 024000
摘要:
本研究旨在探讨化合物WS090152(分子式C28H35NO4Cl4)改善小鼠脂肪肝的作用及其作用机制。以高脂饲料诱导C57BL/6J小鼠形成单纯性脂肪肝(non-alcoholic fatty liver, NAFL)模型,灌胃给予小分子化合物WS090152(50 mg·kg-1·d-1)。以肝脏甘油三酯(triglyceride, TG)含量及病理分析评价肝脏脂质堆积程度;以体重、血清总胆固醇(total cholesterol, TC)和血清TG含量评价全身脂代谢情况;以正糖钳实验葡萄糖输注速率(glucose infusion rate, GIR)评价小鼠的胰岛素敏感性;以Western blot法分析肝组织的蛋白表达;以对基因重组人蛋白酪氨酸磷酸酶1B (human protein tyrosine phosphatase 1B, hPTP1B)活性及对动物肝脏PTP1B蛋白表达的影响评价其对分子靶点PTP1B的作用。结果显示,化合物WS090152能够降低NAFL小鼠肝TG含量(P<0.05)、改善脂肪肝的病理变化(P<0.001),增加机体胰岛素敏感性(P<0.01),降低血TC (P<0.01)和血TG (P<0.05)水平;显著抑制基因重组hPTP1B酶活性(IC50=0.34 μmol·L-1),下调模型小鼠肝脏PTP1B的表达(P<0.05),上调其下游胰岛素受体(insulin receptor, IR)和AKT的磷酸化的水平;降低肝脏内脂质合成相关的因子固醇调节元件结合蛋白1c (lipogenesis-related proteins-1c, SREBP-1c)、脂肪酸合成酶(fatty acid synthase, FAS)和乙酰辅酶A羧化酶(acetyl-CoA carboxylase, ACC)的表达。提示化合物WS090152可能通过抑制分子靶点PTP1B,增加胰岛素敏感性、降低肝脏脂质合成,从而发挥治疗NAFL的作用。
关键词:    C28H35NO4Cl4      单纯性脂肪肝      蛋白酪氨酸磷酸酶1B      胰岛素抵抗     
Investigating the effects of compound WS090152 on non-alcoholic fatty liver in mice
ZHOU Ying1, LI Xiu-li2, YI Qin1, ZHANG Xiao-lin1, TIAN Jin-ying1, WANG Dong-mei1, WU Song1, YE Fei1
1. Beijing Key Laboratory of New Drug Mechanisms and Pharmacological Evaluation Study, State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China;
2. Medical College of Chifeng University, Chifeng 024000, China
Abstract:
To investigate the effects and the mechanism of compound WS090152 on non-alcoholic fatty liver (NAFL), the compound was administrated in C57BL/6J mice fed a high fat diet at 50 mg·kg-1 by lavage. The lipid accumulation in liver was determined by the content of hepatic triglyceride (TG) and the histological pathological analysis. The levels of body weight gain, serum total cholesterol (TC) and TG were measured to evaluate lipid metabolism. Insulin sensitivity was determined by glucose infusion rate (GIR) value in hyperinsulinemic-euglycemic clamp test. The expression of related proteins in liver was measured by Western blot. The effect on the target protein tyrosine phosphatase 1B (PTP1B) was assessed by the activity of recombinate human PTP1B in vitro, and by the expressions of PTP1B in vivo, respectively. The content of hepatic TG (P<0.05) and the pathological changes of the livers (P<0.001) were attenuated, insulin resistance was improved (P<0.01), and the levels of serum TC (P<0.01) and serum TG (P<0.05) were reduced by WS090152 treatment in the mice. The recombinant hPTP1B activity was significantly inhibited with IC50 value of 0.34 μmol·L-1; the expression of PTP1B was significantly downregulated, and the phosphorylation of its downstream insulin receptor (IR) and AKT was upregulated by WS090152 administration in the livers of NAFL mice. The expression of hepatic lipogenesis-related proteins-1c (SREBP-1c), fatty acid synthase (FAS), and acetyl-CoA carboxylase (ACC) was attenuated. These results suggest that compound WS090152 can ameliorate NAFL by increasing insulin sensitivity and decreasing hepatic lipogenesis probably through inhibition of PTP1B.
Key words:    compound C28H35NO4Cl4    non-alcoholic fatty liver    protein tyrosine phosphatase 1B    insulin resistance   
收稿日期: 2016-04-08
DOI: 10.16438/j.0513-4870.2016-0329
基金项目: 国家"重大新药创制"科技重大专项(2012ZX09103-101-063,2012ZX09301002-004);天然药物活性物质与功能国家重点实验室开放基金(GTZK201512).
通讯作者: 叶菲,Tel/Fax:86-10-83150495,E-mail:yefei@imm.ac.cn
Email: yefei@imm.ac.cn
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参考文献:
[1] Bedogni G, Nobili V, Tiribelli C, et al. Epidemiology of fatty liver:an update[J]. World J Gastroenterol, 2014, 20:9050-9054.
[2] Rahimi RS, Landaverde C. Nonalcoholic fatty liver disease and the metabolic syndrome:clinical implications and treatment[J]. Nutr Clin Pract, 2013, 28:40-51.
[3] Ameer F, Scandiuzzi L, Hasnain S, et al. De novo lipogenesis in health and disease[J]. Metabolism, 2014, 63:895-902.
[4] Brumbaugh DE, Friedman JE. Developmental origins of nonalcoholic fatty liver disease[J]. Pediatr Res, 2014, 75:140-147.
[5] Bakke J, Haj FG. Protein-tyrosine phosphatase 1B substrates and metabolic regulation[J]. Semin Cell Dev Biol, 2015, 37:58-65.
[6] Agouni A, Mody N, Owen C, et al. Liver-specific deletion of protein tyrosine phosphatase (PTP) 1B improves obesity- and pharmacologically induced endoplasmic reticulumstress[J]. Biochem J, 2011, 438:369-378.
[7] Ye F, Tao RY, Cong WN, et al. Utilization of fluorescence tracer in hyperinsulinemic-euglycemic clamp test in mice[J]. J Biochem Biophys Methods, 2008, 70:978-984.
[8] Ma YM, Tao RY, Liu Q, et al. PTP1B inhibitor improves both insulin resistance and lipid abnormalities in vivo and in vitro[J]. Mol Cell Biochem, 2011, 357:65-72.
[9] Dietrich P, Hellerbrand C. Non-alcoholic fatty liver disease, obesity and the metabolic syndrome[J]. Best Pract Res Clin Gastroenterol, 2014, 28:637-653.
[10] Basaranoglu M, Kayacetin S, Yilmaz N, et al. Understanding mechanisms of the pathogenesis of nonalcoholic fatty liver disease[J]. World J Gastroenterol, 2010, 16:2223-2226.
[11] Carazo A, León J, Casado J, et al. Hepatic expression of adiponectin receptors increases with non-alcoholic fatty liver disease progression in morbid obesity in correlation with glutathione peroxidase 1[J]. Obes Surg, 2011, 21:492-500.
[12] Asrih M, Jornayvaz FR. Metabolic syndrome and nonalcoholic fatty liver disease:is insulin resistance the link?[J]. Mol Cell Endocrinol, 2015, 418:55-65.
[13] Cong WN, Tao RY, Tian JY, et al. The establishment of a novel non-alcoholic steatohepatitis model accompanied with obesity and insulin resistance in mice[J]. Life Sci, 2008, 82:983-990.
[14] Pansuria M, Xi H, Li L, et al. Insulin resistance, metabolic stress, and atherosclerosis[J]. Front Biosci, 2012, 4:916-931.
[15] Gruben N, Shiri-Sverdlov R, Koonen DP, et al. Nonalcoholic fatty liver disease:a main driver of insulin resistance or a dangerous liaison?[J]. Biochim Biophys Acta, 2014, 1842:2329-2343.
[16] Feldhammer M, Uetani N, Miranda-Saavedra D, et al. PTP1B:a simple enzyme for a complex world[J]. Crit Rev Biochem Mol Biol, 2013, 48:430-445.
[17] Xiao X, Song BL. SREBP:a novel therapeutic target[J]. Acta Biochim Biophys Sin, 2013, 45:2-10.
[18] Tsukamoto H, She H, Hazra S, et al. Fat paradox of steatohepatitis[J]. J Gastroenterol Hepatol, 2008, 23:S104-S107.
[19] Yan F, Wang Q, Lu M, et al. Thyrotropin increases hepatic triglyceride content through upregulation of SREBP-1c activity[J]. J Hepatol, 2014, 61:1358-1364.
[20] Fu Y, Luo N, Klein RL, et al. Adiponectin promotes adipocyte differentiation, insulin sensitivity, and lipid accumulation[J]. J Lipid Res, 2005, 46:1369-1379.
[21] Sanderson SO, Smyrk TC. The use of protein tyrosine phosphatase 1B and insulin receptor immunostains to differentiate nonalcoholic from alcoholic steatohepatitis in liver biopsy specimens[J]. Am J Clin Pathol, 2005, 123:503-509.
[22] Chen PJ, Cai SP, Huang C, et al. Protein tyrosine phosphatase 1B (PTP1B):a key regulator and therapeutic target in liver diseases[J]. Toxicology, 2015, 337:10-20.
[23] Ugi S, Shi K, Nishio Y, et al. Membrane localization of protein-tyrosine phosphatase 1B is essential for its activation of sterol regulatory element-binding protein-1 gene expression and consequent hypertriglyceridaemia[J]. J Biochem, 2009, 146:541-547.
[24] Shimizu S, Ugi S, Maegawa H, et al. Protein-tyrosine phosphatase 1B as new activator for hepatic lipogenesis via sterol regulatory element-binding protein-1 gene expression[J]. J Biol Chem, 2003, 278:43095-43101.