Troglitazone对人腹膜间皮细胞TGF-β_1和纤维连接蛋白表达的影响

Inhibitive effect of troglitazone on TGF-β_1 and fibronectin expression in human peritoneal mesothelial cells

目的:研究过氧化物增生体激活受体γ(PPAR-γ)激动剂troglitazone(曲格列酮,TGZ)对高浓度葡萄糖刺激下体外培养的人腹膜间皮细胞(HPMCs)中TGF-β1和细胞外基质纤维连接蛋白(Fn)表达的影响。方法:采用胰蛋白酶消化法从人大网膜组织中分离间皮细胞,建立稳定的体外培养模型。根据细胞增殖与毒性实验选择troglitazone的最佳浓度15μmol/L,干预高浓度葡萄糖(30mmol/LD-葡萄糖)刺激下的人腹膜间皮细胞。采用逆转录多聚酶链式反应(RT-PCR)半定量检测HPMCs中PPAR-γ,TGF-β1以及Fn的 mRNA表达;采用双抗夹心法酶联免疫吸附实验检测HPMCs培养液中TGF-β1蛋白质水平;Western印迹检测Fn蛋白质水平。结果:高糖(30mmol/LD-葡萄糖)刺激可在 mRNA和蛋白质水平明显上调HPMCs表达TGF-β1和Fn(P<0.01),troglitazone干预高糖孵育的HPMCs,可使TGF-β1和Fn mRNA和蛋白质的表达明显下调(P<0.05)。结论:Troglitazone能够明显抑制在高糖刺激下的HPMCsTGF-β1和Fn的表达,这可能为临床防治长期腹膜透析患者的腹膜纤维化提供一种较为有效的方法。

Objective To investigate the effect of the peroxisome proliferator activated receptor-γ （PPAR-γ） agonist troglitazone on TGF-β1 and fibronectin （Fn） expression in human peritoneal mesothelial cells （HPMCs）. Methods HPMCs were cultured from human omentum by an enzyme digestion method, growing in medium containing 30 mmol/L D-glucose. TGF-β1 and Fn expression were measured in HPMCs in the presence and absence of 15 μmol/L troglitazone. The mRNA expressions of PPAR-γ, TGF-μ1l and Fn were determined by semi-quantification reverse transcriptive PCR （RT-PCR）. The protein of TGF-β1 was determined by enzyme-linked immunosorbent assay （ELISA） and proteins of PPAR-γ and Fn were determined by Western blot. Results The mRNA and protein expression of TGF-β1 and Fn were significantly increased in HPMCs stimulated with 30 mmol/L D-glucose compared with the control group with F12 media （P 〈 0.01 ）. Obvious decrease of TGF-β1 was found in troglitazone （ 15 μmol/L） treated group compared with group stimulated with 30 mmol/L D-glucose （ P 〈 0.05 ）. Exposure of HPMCs to troglitazone reduced the Fn secretion （ P 〈 0.05 ）. conclusion Troglitazone reduced the expressin of TGF-β1 in HPMCs stimulated by 30 mmol/L D-glucose, and reduced Fn production. PPAR-γ agonists may have a specific role in ameliorating the course of progressive peritoneal fibrosis under long-term peritoneal dialysis states.