TGF-β1过表达介导的放疗诱导成纤维细胞向肌成纤维细胞分化的体外研究

Effect of irradiation induced TGF-β1 overexpression on myofibroblast differentiation of fibroblast

目的:探讨体外培养的皮肤成纤维细胞在直线加速器照射后向肌成纤维细胞的转分化。方法:消化法体外培养F344大鼠皮肤成纤维细胞,传代后,在直线加速器下进行照射处理,照射剂量分别为0、2、4、8Gy。应用实时定量PCR检测照射后1、3、5、7d TGF-β1基因表达,应用Western印迹检测TGF-β1和α-SMA蛋白的表达。采用SPSS10.0软件包对数据进行统计学分析。结果:通过酶消化法成功培养成纤维细胞,照射后,TGF-β1基因和蛋白表达随照射剂量增加而上调,随照射后时间增加而下调。照射后第1d,8Gy剂量组TGF-β1表达量最高(P<0.05)。α-SMA蛋白的表达趋势与TGF-β1一致。结论:成纤维细胞照射后转变为肌成纤维细胞并高表达α-SMA,TGF-β1是成纤维细胞照射后早期表达的指标。

PURPOSE： To investigate the effect of irradiation on skin fibroblast into myofibroblast in vitro.METHODS： Enzyme digestion was used to culture F344 rat skin fibroblast.The fibroblasts underwent irradiation by linear accelerator.The gene expression of TGF-β1 was detected by real-time PCR.The protein expression of TGF-β1 and α-SMA was detected by Western blot.The data was analyzed for ANOVA using SPSS 10.0 software package.RESULTS： Skin fibroblast was successfully cultured using enzyme digestion.The gene and protein expression of TGF-β1 was up-regulated with the increase of irradiation dose.However,the expression of the gene was down-regulated with the increase of time.The expression of α-SMA was consistent with TGF-β1.CONCLUSION： The fibroblast was transdifferentiated into myofibroblast after irradiation.TGF-β1 was the early parameter of fibroblast irradiation.