秋水仙碱对二氧化硅致大鼠肺成纤维细胞激活效应的抑制作用

Inhibitory effect of colchicine on silica dioxide-induced rat lung fibroblast activation

目的:观察秋水仙碱对二氧化硅刺激引起的肺成纤维细胞(LF)激活效应的抑制作用。方法:用无血清的二氧化硅培养基培养巨噬细胞(AM)24h,离心收集培养液上清,加入秋水仙碱使其终浓度分别为20、40和80μmol/L。大鼠LF分为秋水仙碱组、阳性对照组和阴性对照组,分别给予低、中、高浓度的秋水仙碱、含二氧化硅的AM培养液上清和不含二氧化硅的AM培养液上清,培养24h。采用Realtime-PCR检测各组LF中α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(COLⅠ)及Ⅲ型胶原(COLⅢ)的mRNA水平,ELISA法测定各组LF培养液上清中COLⅠ、COLⅢ和总蛋白的含量。结果:与阳性对照组相比,随秋水仙碱浓度增高,LF的α-SMA、COLⅠ和COLⅢmR-NA的表达水平呈下降趋势,其中高浓度组下降最明显(P<0.05);LF培养液上清中COLⅠ、COLⅢ和总蛋白含量逐渐降低,其中高浓度组最明显(P<0.05)。结论:秋水仙碱可抑制COLⅠ、COLⅢ的表达,其对二氧化硅诱导大鼠LF的激活效应有抑制作用。

Aim：To observe the inhibitory effect of colchicine on silica dioxide-induced rat lung fibroblast（LF）activation.Methods：AM was stimulated for 24 hours with silica dioxide suspension,then the supernatant was collected and added colchicine,and the concentration was adjusted to 20,40,and 80 μmol/L.The rat LF was prepared and allocated into 5 groups.The positive control was treated without colchicine.AM supernatant without silica dioxide was used in negative control.The other 3 groups were treated with low,mediated and high-dose colchicine.After 24 h,the mRNA expression levels of the α-SMA,COLⅠ and COL Ⅲ were detected by using RT-PCR,and the protein levels of the COLⅠ,COL Ⅲ and total protein were examined by ELISA.Results：Compared with the positive control,the mRNA expression levels of α-SMA,COLⅠand COL Ⅲ in LF were decreased with the increasing concentration of colchicine,and the high-dose colchicine group changed most significantly（P0.05）,so were the content of COLⅠ,COL Ⅲ and total protein in LF culture fluid supernatant（P0.05）.Conclusion：Colchicine could decrease the levels of COLⅠand COL Ⅲ and inhibit the effect of LF stimulated by silica dioxide.