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

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SiO_2体外刺激18 h收集培养上清,与人胚肺FB共同孵育6、12、18、24、36、48、72 h,用~3H-脯氨酸掺入检测48 h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SiO_2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48 h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SiO_2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。

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

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 detected collagen anabolism and secretion with (3)H-proline detected the expression of the procollagen type I in the fibroblast with immunological method detected the quantity of collagen Type I in FB supernatant with Western blot. RESULTS: The anabolism and secretion of collagen were increased in cultured supernatant of silicotic AM exposed to SiO(2), Along with the time, the expression of collagen type I increased. In cultured supernatant of silicotic AM exposed to SiO(2), ((3)H-proline: 1096.500 +/- 76.400, 707.000 +/- 62.160, OD: 0.314 +/- 0.011, OD: 14.218 +/- 0.342. CONCLUSION: SiO(2) may affect the expression of collagen through AM mediation and participate in the formation of lung fibrosis.