干扰素对二氧化硅致肺成纤维细胞激活的抑制效应

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。

Inhibitory effect of interferon for silica-induced stimulation in rat lung fibroblast

OBJECTIVE To evaluate the inhibitive effect of interferon in rat lung fibroblast(LF)exposed to silica.METHODS Silica suspension was used to stimulate AM for 24 hours.Then centrifuge was used to remove silica and collect supernatant.LF was stimulated by the collected supernatant for 24h with interferon(10 U/ml,100/ml and 1 000 U/ml).The positive control was treated without interferon.AM supernatant without silica was used in negative control group.The mRNA expression of the α-smooth muscle actin(α-SMA),(collagenⅠ)COL Ⅰ and(collagen Ⅲ)COL Ⅲ were detected by using real-time PCR,and the protein level of the COL Ⅰ and COL Ⅲ was examined by ELISA.RESULTS Compared to positive control group,the mRNA of α-SMA,collagenⅠ and collagen Ⅲ were decreased in cultured supernatant exposed to interferon at 1 000 u/ml(P﹤0.01).The concentration of collagen,collagen Ⅲ and total protein were decreased compared to positive control(P﹤0.01).CONCLUSION Interferon could inhibit the effect of LF stimulated by silica by decreasing the levels of COL Ⅰ and COL Ⅲ.