氟铝及联合作用对体外培养大鼠破骨细胞数量及骨吸收功能的影响  

Impact of fluorine and aluminum and both action combined on the number of rat osteoclasts and bone resorption cultured in vitro

作　　者：杜光[1] 喻茂娟[1] 徐小雅[1] 金慰芳[1] 高建军[1] 

机构地区：[1]贵阳医学院流行病学研究室,550004

出　　处：《中华地方病学杂志》2013年第4期370-373,共4页Chinese Journal of Endemiology

基　　金：基金项目：科技部国际合作重大项目（2010DFB30530）;贵州省自然科学基金项目（黔科合J字[200912161号）;贵阳市大学生创业基金（2008）

摘　　要：目的观察单纯氟、铝及氟铝联合作用对体外培养大鼠破骨细胞功能的影响，探讨其作用机制。方法机械分离法作用于新生SD大鼠四肢长骨，于TCl99培养液（含10％胎牛血清）中获得破骨细胞和骨髓基质细胞，将破骨细胞接种于96孔培养板和象牙薄切片培养，骨髓基质细胞接种于6孔培养板培养。2h后换液并染毒（氟化钠和氯化铝），分为对照组（氟化钠和氯化铝浓度均为0mol／L）、氟组（氟化钠1．0×10^-4mol／L）、铝组（氯化铝1．0×10。mol／L）和氟铝联合组（氟化钠1．0×10^2mol／L，氯化铝1．0×10^-5mol／L）。培养5d后，对培养板中破骨细胞进行抗酒石酸酸性磷酸酶（TRAP）染色，光镜下检查破骨细胞数目；象牙薄切片经l％甲苯胺蓝染色，光镜下分析破骨细胞骨吸收陷窝面积。骨髓基质细胞染毒培养8h后提取总RNA，实时荧光PCR法测定骨保护素（OPG）和细胞核因子KB受体活化因子配体（RANKL）的表达量水平。结果①氟、铝、氟和铝的交互作用对破骨细胞数量均有影响（F值分别为7．15、6．56、7．98，P均〈0．05），氟组、铝组及氟铝联合组破骨细胞数量[（136．9±22．99）、（135．4±23．5）、（163．0±24．4）个／孔]均高于对照组[（92．5±22．1）个／孔，P均〈0．05]，氟铝联合组高于氟组和铝组（P均〈0．05）。②氟组、铝组及氟铝联合组对破骨细胞骨吸收陷窝面积均有影响（F值分别为10．47、12．64、14．29，P均〈0．05），氟组、铝组及氟铝联合组骨吸收陷窝面积[（0．242±0．031）、（0．293±0．026）、（0．333±0．016）mm2／片]均高于对照组[（0．088±0．030）mm2／片，P均〈0．05]，氟铝联合组高于氟组和铝组（P均〈0．05）。③氟、铝、氟和铝的交互作用对RANKL／OPGmRNA表达量的比值均有影响（F值分别为8．15、15．38、23．59，P均〈0．05），氟组、铝组、氟铝Objective To determine the impact of fluorine and aluminum, and both action combined on the number of rat osteoclasts and bone resorption cultured in vitro and to explore its mechanisms. Methods The osteoclasts and bone marrow stromal cells （BMSCs） isolated from long bone of new born rats were cultured, respectively, in TC199 medium （containing 10% fetal bovine serum） with fluoride, aluminum and fluoride combined with aluminum. The osteoclasts were inoculated in 96-well culture plate and ivory slice, BMSCs in 6-well culture plate, and culture medium was changed after 2 hours incubation. The cells were divided into control group,fluoride group, aluminum group and fluoride combined with aluminum group; the doses of sodium fluoride were 0, 1.0×10^-4, 0, 1.0 ×10^-4 mol/L and the doses of aluminum chloride were 0, 0, 1.0×10^-5, 1.0×10^-5 tool/L, respectively. Tartrate-resistant acid phosphatase （TRAP） staining positive cells were counted under light microscope after TRAP staining on the 5th day and the pit formed in ivory slices were measured by histomorphometry after staining with toludine blue. The expression of osteoprotegerin（OPG） and receptor activator of nuclear factor kappa-B ligand （RANKL） was detected by real-time fluorescence quantitative PCR in BMSCs after 8 h treatment. Results （±） Fluoride, aluminum and the interactive effects of fluoride and aluminum all had impact on the numbers of osteoclasts（F = 7.15, 6.56 and 7.98, respectively, all P 〈 0.05）. The numbers of osteoclasts in fluoride group, aluminum group and fluoride combined with aluminum group [ （136.9 ± 22.99）, （135.4 ± 23.5）, （163.0 ± 24.4） per well] were higher than that in the control group[ （92.5 ± 22.1） per well, all P 〈 0.05]. （2） Fluoride, aluminum and the interactive effects of fluoride and aluminum all had impact on the resorption pit area on ivory slices（F = 10.47, 12.64, 14.29, respectively, all P 〈 0.05）. The resorption pit area on ivory slices in fluoride group, aluminu

关 键 词：氟化物 铝化物 破骨细胞 细胞核因子Kp受体活化因子配体 骨保护素 细胞 培养技术 

分 类 号：R114[医药卫生—卫生毒理学]