摘要
目的观察不同剂量氟化物在不同时间段对成纤维细胞和成骨细胞核心结合因子α1(cbfa1)表达的影响。方法采用细胞培养的方法,将细胞分为对照组和6个染氟组(0．1、1．0、100．0、1000．0、10 000．0、20 000．0μg／L),分别在5个染氟时间段(2、4、24、48、72 h)收集培养细胞培养上清液和细胞,应用酶联免疫吸附(ELISA)法和免疫组化方法检测cbfa1蛋白含量和表达,应用RT-PCR方法检测染氟48 h cbfa1 mRNA的表达。结果①与对照组比较(2．13±0．07),成纤维细胞染氟48 h时,cbfa1蛋白含量在0．1、1．0、100．0、1 000．0、20 000．0μg／L组[(2.35±0．08)、(2．28±0．09)、(2．32±0．09)、(2．25±0．08)、(2．28±0．09)]及染氟72 h的10 000．0μg／L组(2．48±0．22)明显升高;染氟48 h,cbfa1 mRNA表达呈升高趋势,其中10 000．0μg／L组(1．29±0．30)与对照组(1．02±0．12)比较,明显增强;免疫组化结果显示,染氟48 h的0．1μg／L组成纤维细胞内可见明显cbfa1阳性表达。②在成骨细胞,染氟组cbfa1蛋白含量较对照组有不同程度增加;染氟48 h时,0．1、1．0、100．0、1 000．0μg／L组cbfa1 mRNA表达较对照组(1．27±0．20)升高,其中0．1μgL组(1．34±0．19)明显升高。结论氟能提高成纤维细胞和成骨细胞cbfa1蛋白含量,促进cbfa1和cbfa1 mRNA表达,成纤维细胞可能在氟骨症骨周化骨的发生中起重要作用。
Objective To study the effects of fluoride on the expression of Core-binding facor αl （cbfal） in fibroblast and osteoblast. Methods Osteoblast and tbroblasts were exposed to different concentrations of fluoride （F- 0,0.1,1.0,100.0,1 000.0,10 000.0,20 000.0 μg/L）. The levels of cbfal mRNA and protein at certain time were measured by using RT-PCR,ELISA and immunohistochemistry（IHC） methods. Results ①In fibroblast, the content of cbfal protein increased obviously in groups of 0.1,1.0,100.0,10 000.0 and 20 000.0 μg/L at 48 h[ （2.35 ± 0.08）, （2.28 ± 0.09）, （2.32 ± 0.09）, （2.25 ± 0.08）, （2.28 ± 0.09）] and group of 10 000.0 μg/L at 72 h （2.48 ± 0.22）. The level of cbfal mRNA was much higher in the group of 10 000.0 μg/L at 48 h（1.29 ± 0.30） than it was in the control （1.02 ± 0.12）. The stronger expression of cbfal was observed by IHC in fibroblast treated with 0.1μg/L for 48 h.②Comparing to the control,the content of cbfal protein increased in most of groups of fluoride-treated osteoblast. The expression of cbfal mRNA were higher in the groups of 0.1,1.0,100.0,1 000.0 μg/L and much higher in group of 0.1 μg/L （1.34 ± 0.19） than it was in the control （1.27 ± 0.20）. Conclusions The expression of both mRNA and protein of cbfal were increased significantly in fibroblast and osteoblast treated with fluoride. This study suggested that fibroblast possibly play important role in the developing of extraperiosteal ossification of skeletal fluorosis.
出处
《中国地方病学杂志》
CAS
CSCD
北大核心
2006年第6期629-632,共4页
Chinese Jouranl of Endemiology
基金
国家自然科学基金资助项目(30471498)
