细胞直接共培养模型在大鼠肺成纤维细胞转分化中的应用

目的:探讨细胞直接共培养模型在大鼠肺成纤维细胞转分化研究中的应用价值。方法:分离SD大鼠肺巨噬细胞和成纤维细胞,采用ThinCertTM进行共培养,并分为4组:对照组加入无SiO2粉尘的培养基,SiO2低、中、高剂量处理组分别加入终质量浓度为25、50和100mg/L的SiO2粉尘悬液,培养24h后,收集成纤维细胞和培养上清,分别采用real-time PCR检测成纤维细胞中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法测定培养上清中α-SMA、COLⅠ和COLⅢ蛋白。结果:SiO2处理组的α-SMA、COLⅠ和COLⅢmRNA和蛋白的表达水平明显高于对照组,且随SiO2质量浓度的升高,α-SMA、COLⅠ和COLⅢmRNA和蛋白的表达水平有升高的趋势(P<0.05)。结论:细胞直接共培养模型可用于大鼠肺成纤维细胞转分化的研究。

Application of cell co-culture in lung fibroblast transdifferentiation

Aim:To evaluate the application value of cell co-culture model in the study of the lung fibroblast(LF) transdifferentiation.Methods:The primary macrophages and LF of SD rats were isolated and co-cultured using the ThinCertTM.0,25,50,and 100 mg/L silica dioxide suspension was added to stimulate macrophages for 24 h.LFs and the culture supernatant were collected,and the mRNA and protein expression levels of the α-smooth muscle actin(α-SMA),collagenⅠ(COLⅠ) and collagen Ⅲ(COLⅢ) were detected by using real-time PCR and ELISA,respectively.Results:Compared with control,the mRNA and protein level of α-SMA,COLⅠand COLⅢ were increased in silica dioxide treatment groups,which increased with the concentration of silica dioxide(P<0.05).Conclusion:Cell co-culture model could be applied to study the LF transdifferentiation.