氟对大鼠成骨细胞Wnt3α、β-链蛋白mRNA和蛋白表达的影响  

Effect of fluoride on expression of mRNA and protein of Wnt3a and β-catenin in osteoblast of rats

作　　者：陈锡山[1] 于燕妮[1] 易韦[2] 万良斌[1] 谢莹[1] 

机构地区：[1]贵阳医学院病理学教研室,550004 [2]贵州省人民医院病理科

出　　处：《中华地方病学杂志》2013年第2期140-145,共6页Chinese Journal of Endemiology

基　　金：国家自然科学基金（81260419）;科技部国际合作项目（2010DFB30530）;贵州省科技厅社会发展攻关项目（黔科合SY[2011]3006号）

摘　　要：目的观察氟中毒大鼠成骨细胞Wnt3α、β-链蛋白（catenin）mRNA和蛋白表达，探讨氟骨症发生与Wnt通路的关系。方法健康SD大鼠36只，体质量100—120g，按体质量将大鼠随机分为3组，每组12只。对照组大鼠饮用自来水（含氟量〈1mg／L），低氟组、高氟组大鼠分别饮用含5、50mg／L氟化钠的自来水。大鼠饲养8个月，建立慢性氟中毒模型。饲养期间检查大鼠氟斑牙发生情况，股动脉放血处死大鼠前收集大鼠24h尿样，处死后取股骨组织。采用氟离子选择电极法测定尿氟和骨氟含量；固相夹心酶联免疫吸附（ELISA）法测定血清骨碱性磷酸酶（BALP）和抗酒石酸酸性磷酸酶一5b（TRACP一5b）含量；光镜下观察骨组织的形态学变化，测量骨皮质厚度、骨小梁宽度及密度变化；原位杂交技术和免疫组化方法检测成骨细胞Wnt3α、β-cateninmRNA和蛋白表达。结果大鼠氟斑牙检出率低氟组为66．7％（8／12），高氟组为91．7％（11／12），对照组为0．O％（0／12），组间比较差异有统计学意义（x。：21．6，P〈0．05）。尿氟和骨氟组问比较差异有统计学意义（F=36．57、467．02，P均〈0．05），其中低氟组[（2．06±0．64）mg／L、（632．33±123．21）mg／kS]和高氟组[（7．69±1．96）mg／L、（1088．75±156．16）mg／kg]高于对照组[（1．26±0．17）mg／L、（305．58±91．26）mg／kg，P均〈0．05]，高氟组高于低氟组（P均〈0．05）。血清BALP和TRACP-5b组间比较差异有统计学意义（F=89．57、7．68，P均〈0．05）。其中低氟组[（31．47±5．30）U／L]和高氟组[（54．61±2．27）U／L]，血清BALP明显高于对照组[（16．24±1．57）U／L，P均〈0．05]，高氟组高于低氟组（P〈0．05）；血清TRACP-5b，低氟组[（3．45±1．85）U／L）明显高于对照组[（1．264±0．23）U／L]和高氟组[（2．744±1．8）]U／L，P均〈0．05]。光镜下，高Objective To explore the effect of excessive fluoride on expression of mRNA and protein of Wnt3a and β-catenin in rats＇ osteoblasts and its correlation with pathogenic mechanism of fluorosis. Methods Thirty-six healthy SD rats, weighting 100 - 120 g and according to body mass, were randomly divided into threegroups（twelve in each group）. The rats of control were fed with tap water（fluoride 〈 1 mg/L） and the experimental rats were exposed to NaF （low-fluoride group ： 5 mg/L, high-fluoride group ： 50 mg/L） added to the drinking water to establish the chronic fluorosis model. After fed for eight month, all rats were killed and metaphysic of femoral was collected. Rat dental fluorosis was observed and bone fluorine was detected by ashing-fluorin ion selective electrode method. The content of bone alkaline phosphatase （BALP） and tartrate-resistant acid phosphatase 5b（TRACP 5b） in rats＇ serum was detected by enzyme-linked immunosorbent assay（ELISA）. The morphologic changes of the bone were observed by microscopy. The expression of mRNA and protein of Wnt3a and β-catenin in osteoblasts of rats was analyzed with gray scale by hybridization in situ and immunohistochemistry methods, respectively. Results Detection rate of dental fluorosis, fluoride contents of urine and bone were significantly increased[control group： 0.0%, （1.26 ＋ 0.17）mg/L, （305.58 ± 91.26）mg/kg; low-fluoride group： 66.7%, （2.06 ± 0.64）mg/L, （632.33 ± 123.21）mg/kg; high-fluoride group： 91.7%, （7.69 ± 1.96）mg/L, （1088.75 ± 156.16） mg/kg] in the rats treated with fluoride, the difference between groups was statistically significant（±2 = 21.6; F = 36.57, 467.02; all P 〈 0.05）. The contents of BALP and TRACP-5b in rats＇ serum were significantly different between groups（F = 89.57, 7.68 ; all P 〈 0.05）. Compared with control group [ （16.24 ± 1.57）U/L], the contents of BALP in rats＇ serum of the low-fluoride and high-fluoride groups[ （31.47 ± 5.30） and （54.61 �

关 键 词：大鼠 氟中毒 成骨细胞 WNT3A Β-链蛋白 

分 类 号：R114[医药卫生—卫生毒理学]