氟化物对成纤维细胞和成骨细胞骨形态发生蛋白-2表达的影响

Effect of fluoride on the expression of Bone morphogenetic protein 2 in fibroblasts and osteoblasts

目的观察氟化物对成纤维细胞和成骨细胞骨形态发生蛋白-2（BMP-2）表达的影响。方法采用体外细胞培养的方法，将细胞分为对照组和6个染氟（F^-，0．1、1．0、100．0、1000．0、10000．0、20000．0μg／L）组，分别在5个染氟时间段（2、4、24、48、72h）收集培养上清液和细胞，应用酶联免疫吸附法（ELISA）和免疫组化方法检测BMP-2蛋白，RT-PCR方法检测染氟48hBMP-2mRNA的表达。结果与对照组比较．成纤维细胞染氟48hBMP-2mRNA表达呈普遍增强趋势，但差异无统计学意义（P〉0．05）；免疫组化检查，可见染氟0．1、1．0、1000．0μg／L组成纤维细胞BMP-2阳性着色较对照组增强；染氟72h，培养上清中BMP-2蛋白在1．0、100．0、20000．0μg／L组分别为（0．11±0．01）、（0．11±0．01）、（0．11±0．01），与对照组（0．07±0．01）比较有明显增高（P〈0．05）。与染氟成纤维细胞比较，染氟成骨细胞BMP-2mRNA和蛋白表达升高出现早．持续时间较长．增强程度更为明显。结论成纤维细胞和成骨细胞在氟化物的刺激下，BMP-2mRNA和蛋白表达有所升高，推测BMP-2可能是氟化物诱导成纤维细胞中成骨细胞核心结合因子α1（cbfa1）和骨钙素（OCN）表达的重要中介环节。

Objective To study the effects of fluoride on the expression of Bone morphogenetic protein 2 （BMP-2） in fibroblast and osteoblast. Methods Fibroblasts and osteoblasts were exposed to a series of concentrations of fluoride 0, 0.1,1.0,100.0,1 000.0,10 000.0,20 000.0 μg/L, respectively. At the time of 2,4,24,48 and 72 h, the levels of BMP-2 mRNA and protein at certain time were measured using RT-PCR, ELISA and immunohistochemistry （IHC） method. Results In fibroblasts, the content of BMP-2 protein increased obviously in fluoride of 1.0, 100.0 and 20 000.0 μg/L at 72 h compared with control group （P 〈 0.05）. A strong expression of BMP-2 was observed in fibroblasts treated with fluoride of 0.1, 1.0 and 1 000.0 μg/L for 48 h, but without statistical difference. The expression of BMP-2 mRNA were higher in fluoride groups than it was in control group. While in osteoblast, the expression of BMP-2 induced by fluoride were much earlier, longer and stronger. Conclusions The higher expression of both mRNA and protein of BMP-2 induced by fluoride may play an important role in enhancing the levels of Core-binding factor α1 （cbfal） and oeteocalcin and further in promoting osteogenetic function in fibroblast.