氟对成纤维细胞和成骨细胞TGF-β和Smad2/3表达的影响

Effects of fluoride on expressions of TGF-β and Smad2/3 in fibroblasts and osteoblasts

目的：观察不同浓度氟化物在不同时间段对成纤维细胞和成骨细胞中转化生长因子β（TGF-β）和Smad2／3表达的影响。方法：将成纤维细胞和成骨细胞各分7个染氟组（0、0．0001、0．001、0．1、1、10和20mg·L^-1），在5个染氟时间段（2、4、24、48和72h）采集细胞培养上清，应用酶联免疫吸附（ELISA）法检测TGF—β和Smad2／3含量，应用RT—PCR方法检测染氟48h细胞TGF-β mRNA的表达。结果：与染氟0mg·L^-1组比较，氟作用2h的0．001、0．1、1、10和20mg·L^-1组和氟作用4h的0．0001、0．001、0．1、10和20mg·L^-1组成纤维细胞TGF—β蛋白表达明显降低（P〈0．01或P〈0．05）；染氟48h时1和20mg·L^-1组成纤维细胞TGF—β mRNA表达有所增强，染氟2～24h时Smad2／3表达多呈上升趋势，但差异无显著性（P〉0．05）；成骨细胞染氟48h时0．0001、0．001和1mg·L^-1组TGF—β蛋白表达增强（P〈0．01或P〈0．05），TGF-β mRNA的表达呈增强趋势；染氟成骨细胞Smad2／3表达多呈上升趋势，其中染氟48h的20mg·L^-1组表达明显升高（P〈0．01）。结论：染氟对成纤维细胞和成骨细胞TGF—β表达具有不同的作用特点，TGF—β在成纤维细胞成骨表型转换过程中可能起重要作用。

Objective To explore the effects of fluoride with different doses on the expressions of TGF-β and Smad2/3 in fibroblast and osteoblast at different periods. Methods Fibroblasts and osteoblasts were exposed to different concentrations of fluoride （0, 0.0001, 0.001, 0.1, 1, 10 and 20 mg· L^-1） . The levels of TGF-β protein and Smad2/3 at 2, 4, 24, 48 and 72 h after treatment were measured by using ELISA method. The expression of TGF-β mRNA was tested with RT-PCR method. Results In fibroblasts, the contents of TGF-β protein were decreased in the groups of 0. 001, 0.1, 1, 10 and 20mg· L^-1 F^- at the time of 2 h and in the groups of 0.0001, 0.001, 0.1, 10 and 20 mg· L^-1 at the time of 4 h （P〈0.01, P〈0.05） . The levels of TGF-β mRNA were increased in the groups of 1 and 20 mg · L^-1 F^- at 48 h and the expressions of Smad2/3 were increased in fluoride groups during 2 to 24 h. In osteoblasts, the expressions of TGF-β protein and mRNA were increased significantly in the groups of 0. 0001, 0. 001 and 1 mg· L^-1 F^- at 48 h （P〈0. 01, P〈0. 05） and the level of Smad2/3 elevated in the group of 20 mg · L^-1 F^- at 48 h （P〈0. 01） ; Conclusion TGF-β may play roles in the developing of osteogenesitic function in fibroblasts induced by fluoride.