SB-431542抑制转化生长因子β1诱导的人胚肺成纤维细胞(HLF)细胞外基质的合成

SB-431542 inhibits extracellular matrix components deposition induced by transforming growth factor β1 in cultured human lung fibroblasts

目的研究转化生长因子β1(TGFβ1)的特异性小分子拮抗剂SB 431542在TGFβ1信号系统功能增强的状况下,对人胚肺成纤维细胞(HLF)合成细胞外基质的影响。方法人胚肺成纤维细胞系(HLF)于体外正常培养。用MTT法评价SB 431542对HLF的细胞毒作用;以半定量RTPCR(reverse transcription polymerase chain reaction)法和琼脂糖凝胶电泳法观察细胞外基质(ECM)中主要组成分子I型胶原α1(Col Iα1)、纤溶酶原激活物抑制剂1(PAI 1)和纤维粘连素(FN)的变化。结果SB 431542对HLF的LC50(50%lethal concentration)为52μmol.L-1;在TGFβ1(10μg.L-1)诱导下,SB 431542(0.1、0.5、1.0、10μmol.L-1)可剂量依赖性抑制Col Iα1、PAI 1和FN的mRNA在HLF细胞内的转录。结论SB 431542可明显抑制由于肺内TGFβ1信号系统增强而引起的ECM的合成,表明作用于TGFβ1途径的SB 431542或其他小分子化合物可能对防治肺纤维化有效。

Objective To investigate the inhibitory effects of SB-431542, a small molecule antagonist of transforming growth factor-β1 （TGF-β1）which plays a pivotal role in pulmonary fibrogenesis, on extracellular matrix（ECM） production in TGF-β1-treated human lung fibroblasts （HLFs） in vitro. Methods HLF was cultured in routine conditions. The mRNA levels of ECM, including plasminogen activator inhibitor-1 （PAI-1） type I collagen（Col Iα1）, and febronectin （FN） were assessed by reverse transcription polymerase chain reaction （RT-PCR）, respectively. MTT viability assay method was used to assess potential toxicity of SB-431542 to HLFs. Results In TGF-β1-treated HLCs, SB-431542（0.1,0.5, 1.0, 10μmol·L^-1 ） significantly halted the transcription of PAI-1, Col Iα1 and FN. MTT assay showed LC50 values of 52 μmol·L^-1. Conclusions As a selective and specific small molecular inbibitor of type I TGF-β receptor, SB-431542 significantly attenuates ECM components accumulation in cultured HLFs under a fibrogenic conditions, and can be used as a possible candidate for anti-lung fibrosis.