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.