新型PPARγ部分激动剂CMHX003在3T3-L1细胞中的胰岛素增敏作用

Role of CMHX003，a novel peroxisome proliferator-activated receptor γ selective partial agonist in enhancing insulin sensitivity in 3T3-L1 cells

摘要:

目的：探讨噻唑烷二酮类（thiazolidinediones，TZDs）新型衍生物CMHX003在3T3-L1细胞中的过氧化物酶体增殖物激活受体γ（peroxisome proliferator-activated receptor γ，PPARγ）部分激动活性及促进脂肪细胞分化的作用。方法：用双荧光素酶报告基因检测方法测定HEK293细胞中对照组、CHMX003（1 μmol/L）、CHMX003（10 μmol/L）和罗格列酮（rosiglitazone，Rosi）（10 μmol/L）对PPARγ的激动活性。采用经典鸡尾酒诱导法诱导3T3-L1前脂肪细胞分化，诱导过程中分别加入CHMX003（1 μmol/L）、CHMX003（10 μmol/L）和Rosi（10 μmol/L）处理，诱导分化第7天，用油红O染色观察细胞分化情况，real-time PCR检测细胞脂联素和脂肪酸结合蛋白（aP2）基因mRNA表达水平；ELISA法测定细胞培养上清液中脂联素水平。结果：CMHX003对PPARγ的激动活性低于Rosi（2.46±0.08，3.31±0.15，F=132.19，P=0.008）；CMHX003促进3T3-L1前脂肪细胞分化及细胞内脂质沉积的作用较Rosi弱；而增强脂联素基因表达（59.41±3.01，107.91±16.49，χ2=9.031，P=0.112）及增加脂联素分泌水平（74.61±16.94，140.07±30.67，χ2=9.051，P=0.135）的作用和Rosi相似，且较少增强aP2基因的表达（3.07±1.86，75.95±26.04， χ2=8.868，P=0.049）。结论：新型TZDs 衍生物CMHX003具有PPARγ部分激动活性，有望成为安全有效治疗2型糖尿病和代谢紊乱的新型药物。

Abstract:

Objective：To investigate the effect of CMHX003，a novel peroxisome proliferator-activated receptor γ（PPARγ） selective partial agonist，on the enhancement of insulin sensitivity in 3T3-L1 preadipocytes. Methods：PPARγ agonist activity was determined by using luci-ferase reporter gene assay. 3T3-L1 preadipocytes were differentiated into adipocytes according to standard protocols. Oil red O staining was performed at the end of differentiation and the adiponectin and aP2 mRNA levels were detected by real-time PCR. The secreted adiponectin levels in the culture medium were measured with mouse adiponectin ELISA kit. Results：The result of luciferase reporter assay suggested that PPARγ agonist activity of CMHX003 was higher than that of rosiglitazone（2.46±0.08，3.31±0.15，F=132.19，P=0.008）. CMHX003 had weaker role in promoting 3T3-L1 preadipocyte differentiation and triglyceride accu－mulation than rosiglitazone but similar role in increasing expression and secretion of adiponectin（59.41±3.01，107.91±16.49， χ2=9.031，P=0.112）（74.61±16.94，140.07±30.67， χ2=9.051，P=0.135） as rosiglitazone. Meanwhile，CMHX003 was less likely to increase aP2 expression（3.07±1.86，75.95±26.04， χ2=8.868，P=0.049）. Conclusions：CMHX003 may potentially be developed into an effective and safe agent in the treatment of type 2 diabetes mel－litus and metabolic disorders.