矽肺肺泡巨噬细胞对人胚肺成纤维细胞Ⅰ型胶原表达的体外研究

Effect of human silicotic alveolar macrophages on expression of collagen type Ⅰ in human embryonic lung fibroblasts

目的探讨矽肺患者的肺泡巨噬细胞（AM）是否通过影响肺成纤维细胞（FB）胶原的表达，参与矽肺纤维化的发生发展。方法收集矽肺患者AM，经SiO2体外刺激18h收集培养上清，与人胚肺FB共同孵育6、12、18、24、36、48、72h，用^3H-脯氨酸掺入检测48h胶原的合成与分泌，免疫细胞化学方法、Western blot方法检测各时间点Ⅰ型胶原表达。结果经SiO2刺激矽肺患者AM培养上清，可使FB中Ⅰ型胶原的表达增高，实验组48h细胞内、外3H-脯氨酸掺入值分别为1259．500±73．879、790．500±70．315，免疫细胞化学染色的积分光密度值为0．341±0．011，Western blot条带扫描灰度值为14．218±0．342，均明显高于空白对照组及AM对照组，差异有统计学意义（P〈0．05或P〈0．01）。结论SiO2可通过AM介导影响FB中Ⅰ型胶原表达，参与肺纤维化的形成。

Objective To study the effect of the cultured supernatant of human silicotic alveolar macrophages （AM） on the expression of the collagen type Ⅰ in human embryonic lung fibroblasts. Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 18 h. Then the cultured supernatant were used to culture human embryonic lung fibroblasts for 6 h, 12 h, 18 h, 24 h, 36 h, 48 h, 72 h. Then the collagen anabolism and secretion were detected with ^3H- proline; and the expression of the procollagen type Ⅰ in the fibroblast was detected with immunological method. The quantity of collagen Type Ⅰ in FB supernatant was detected with Western blot. Results The anabohsm and secretion of collagen were increased in cultured supernatant of silicotic AM exposed to SiO2, Along with the time, the expression of collagen type Ⅰ increased. In cultured supernatant of silicotic AM exposed to SiO2, （3H-proline：1096.500±76.400,707.000±62.160,OD： 0.314±0.011,OD： 14.218±0.342. Conclusion SiO2 may affect the expression of collagen throgh AM mediation and participate in the formation of lung fibrosis.