氟对成纤维细胞核心结合因子α1及增殖活性量-效关系的影响

Effect of fluoride on the expression of core binding factor alpha 1 in fibroblasts and dose-effect relationship between fluoride and proliferation activity of fibroblasts

背景:氟通过刺激成纤维细胞向成骨细胞方向转化,可能在氟性骨损伤骨周化骨中起重要作用。核心结合因子α1作为成骨细胞特异性转录因子,是成骨细胞分化以及成骨的必要条件。目的:观察不同质量浓度氟化钠干预体外培养新生大鼠皮肤成纤维细胞不同时间后核心结合因子α1的表达及成纤维细胞的增殖活性。方法:采用细胞原代培养方法,将细胞分为对照组和7个染氟组(0.0001,0.001,0.01,0.1,1,10,20mg/L),分别在4个染氟时间段(24,48,72,96h)收集细胞培养上清液,应用酶联免疫法测定核心结合因子α1的含量;采用四甲基偶氮唑盐比色法观察氟对成纤维细胞增殖活性的影响。结果与结论:氟能提高成纤维细胞核心结合因子α1的表达,核心结合因子α1可能在成纤维细胞致氟性骨损伤骨周化骨中起重要作用。氟对成纤维细胞的增殖活性的影响呈一定的剂量-时间-效应关系,短时间-低剂量的氟可提高成纤维细胞的增殖活性,随着染氟剂量的增加及染氟时间的延长,成纤维细胞增殖活性明显减弱。

BACKGROUND： Fluoride may play an important role in extraperiosteal ossification of severe disabled skeletal fluorosis through inducing osteogenic differentiation of fibroblasts. Core binding factor alpha 1 （Cbfal）, as a specific transcription factor of osteoblasts, is a necessary condition for osteoblast differentiation and osteogenesis. OBJECTIVE： To study the changes in Cbfal expression and proliferation activity of fibroblasts when skin fibroblasts from infant rats cultured in vitro were exposed to fluoride at different concentrations and different time. METHODS： Supernatant of control group and seven fluoride exposed groups （0.000 1, 0.001,0.01, 0.1, 1, 10 and 20 mg/L） were collected at 24, 48, 72 and 96 hours after fluoride, and the Cbfal was determined by enzyme linked immunosorbent assay. In addition, for studying the effect of fluoride on proliferation activity of fibroblast, 3-（4,5-Dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide method was employed. RESULTS AND CONCLUSION： The Cbfal of flbroblast was quantitatively increased by fluoride, and played an important part in the developing of extraperiosteal ossification of skeletal fluorosis. The effect of fluoride on proliferation activity of fibroblast presented in a dose-time-effect dependent manner. Shorter time and lower dose of fluoride could enhance this activity; however, as the dose increasing and time expanding, this activity was significantly decreased,