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目的探讨干扰素对二氧化硅(SiO2)刺激引起的肺成纤维细胞激活效应的抑制作用。方法用无血清的SiO2培养基刺激巨噬细胞(AM)24h,离心收集培养液上清。加入干扰素使其终浓度为10U/ml、100U/ml、1000U/ml,分别刺激LF;阳性对照未加干扰素;阴性对照用不含SiO2的AM上清培养肺成纤维细胞(LF)。RT-PCR检测α-平滑肌肌动蛋白(α-smooth muscleactin,α-SMA)、Ⅰ型胶原(collagenⅠ,COLⅠ)及Ⅲ型胶原(collagenⅢ,COLⅢ)的mR-NA水平;ELISA法测定COLⅠ和COLⅢ。结果干扰素浓度为1000U/ml时,α-SMA、COLⅠ和COLⅢ的转录水平为1.17±0.42、1.56±0.13和1.67±0.45,低于阳性对照组的2.97±0.68、4.10±1.42和3.30±0.46,差异有统计学意义(P﹤0.01);浓度为1000U/ml时,COLⅠ、COLⅢ和总蛋白分泌为1.55±0.23、2.27±0.24和2.27±0.24,低于阳性对照组3.79±0.22、3.58±0.55和4.45±0.11,差异有统计学意义(P﹤0.05)。结论干扰素抑制COLⅠ、COLⅢ的表达,说明其对矽尘诱导大鼠肺成纤维细胞的激活效应有抑制作用。
Objective To investigate the inhibitory effect of interferon on the activation of lung fibroblasts induced by silica (SiO2) stimulation. Methods The macrophages (AM) were stimulated with serum-free SiO2 medium for 24 hours, and the supernatant of the medium was collected by centrifugation. Interferon was added to a final concentration of 10 U / ml, 100 U / ml and 1000 U / ml, respectively, to stimulate LF; positive control without interferon; negative control lung fibroblasts (LF) were cultured with SiO2-free AM supernatant. The mR-NA levels of α-smooth muscle actin (α-SMA), collagenⅠ (COLⅠ) and collagen Ⅲ (COLⅢ) were detected by RT-PCR. . Results The transcriptional levels of α-SMA, COLⅠ and COLⅢ were 1.17 ± 0.42, 1.56 ± 0.13 and 1.67 ± 0.45 at the concentration of 1000 U / ml, which were lower than that of the positive control group at 2.97 ± 0.68, 4.10 ± 1.42 and 3.30 ± 0.46 (P <0.01). At the concentration of 1000 U / ml, COL Ⅰ, COL Ⅲ and total protein secretion were 1.55 ± 0.23, 2.27 ± 0.24 and 2.27 ± 0.24, respectively, lower than the positive control group 3.79 ± 0.22,3.58 ± 0.55 and 4.45 ± 0.11, the difference was statistically significant (P <0.05). Conclusion Interferon inhibits the expression of COL Ⅰ and COL Ⅲ, indicating that interferon inhibits the activation of rat lung fibroblasts induced by silica dust.