细胞外信号调节蛋白激酶1/2信号通路对SiO2活化的肺成纤维细胞转化生长因子-β1表达的影响

The effect of extracellual signal-regulated kinase1/2 signaling pathway on the expression of transforming growth factor-β1 in lung flbroblast activated by silicon dioxide

目的 探讨细胞外信号调节蛋白激酶（ERK）信号通路对矽尘诱导的人胚肺成纤维细胞（HELF）中转化生长因子（TGF-β1）表达的影响及调控作用.方法 收集矽肺患者支气管肺泡灌洗液中的肺泡巨噬细胞（AM）,在体外将AM分别用不含SiO2的DMEM培养液和含SiO2（50 μg/ml）的DMEM 培养液培养作用18 h后,将收集的AM条件培养上清液与HELF共同孵育.通过采用免疫细胞化学法检测应用ERK1/2抑制剂PD98059干预后SiO2活化的矽肺患者AM上清介导的HELF中TGF-β1表达的改变.结果 SiO2处理组HELF中TGF-β1的表达量（0.3227±0.0238）明显高于空白对照组（0.1637±0.0196）及AM对照组（0.2406±0.0225）,而PD98059干预组TGF-β1的表达量（0.2711±0.0229）明显低于SiO2处理组,差异均有统计学意义（P＜0.05）.结论 PD98059对由SiO2刺激的矽肺患者AM条件培养上清介导的HELF中TGF-β1的表达有一定抑制作用,ERK信号途径在一定程度上可能介导SiO2刺激的HELF细胞因子表达.

Objective To investigate the effect and regulation of extracellual signal-regulated kinase （ERK1/2）signaling pathway on the expression of transforming growth factor-β1 （TGF-β1） in human embryonic lung fibroblasts induced by SiO2. Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and in the presence or absence of SiO2 （50 ug/ml） exposition for 18h, and then the conditioned supernatants were used to incubate HELF. The expressions of TGF-β1 of the HELF acted with the conditioned AM supernatant fluid were detected with the immunocytochemistry method after treatment with PD98059 of inhibitor of ERK. Results The expression of TGF-β1 in HELF of the SiO2 treatment group（OD value is 0.322 7±0.023 8）exceed blank group （OD value is 0.163 7±0.019 6） and AM control group（OD value is 0.240 6±0.022 5） by the immunocytochemistry method. But the expression of TGF-β1 had reduction in some extent in the PD98059 intervention group （OD value is 0.271 l±0.022 9）. The values were statistically different （P〈0.05）. Conclusion ERK inhibitor PD98059 have inhibition effect on the expression of transforming growth factor-pi and expression of cytokine of human embryonic lung fibroblasts stimulated by SiO2, The study indicate that the proliferation and collagen prodction of HELF activated by SiO2 are mediated by ERK/MAPK signal pathway in some extent. PD98059 may antagonizes silica-induced lung fibrosis by inhibiting the expression of transforming growth factor-β1.