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目的观察慢性氟中毒时大鼠成骨细胞中成骨相关蛋白SPARC mRNA及其蛋白表达水平的影响,并探讨SPARC在氟骨症发病机制中的可能作用。方法 36只健康SD大鼠按体重、性别随机分3组,对照组自由饮用自来水(氟<1mg/L),低、高氟组分别饮用添加氟化钠5、50mg/L的自来水。饲养8个月后,股动脉放血处死大鼠,用氟离子选择电极法测各组尿氟、骨氟含量。HE染色后光镜下观察股骨下段骨组织的病理学改变及形态计量学测量。免疫组织化学和原位杂交技术检测成骨细胞中SPARC蛋白及其mRNA的表达水平。结果低氟组大鼠尿氟、骨氟含量[(6.09±0.56)mg/L、(12.65±3.07)μg/g]显著高于对照组[(1.36±0.51)mg/L、0.67±0.16)μg/g],差异有统计学意义(P均<0.05),高氟组[(7.69±0.64)mg/L、(26.53±5.88)μg/g]较低氟组升高,差异有统计学意义(P均<0.05)。染氟组大鼠股骨下段呈骨质硬化表现。低氟组大鼠SPARC蛋白及其mRNA表达水平(125.24±1.91、100.90±1.13)明显高于对照组(109.70±1.70、88.03±1.26),差异有统计学意义(P均<0.05),高氟组(139.47±1.03、119.22±1.23)较低氟组升高,差异有统计学意义(P均<0.05)。结论过量氟可通过上调大鼠成骨细胞中SPARC蛋白及其mRNA表达水平来调节成骨细胞的增殖、分化、成熟等过程,使骨形成增加,表现为骨硬化型。
Objective To observe the effect of SPARC mRNA and protein expression on osteoblast in rats with chronic fluorosis and to explore the possible role of SPARC in the pathogenesis of skeletal fluorosis. Methods Thirty - six healthy SD rats were randomly divided into three groups according to their body weight and sex. The control group were given free tap water (fluoride <1 mg / L), and the low and high fluoride groups were respectively treated with 5 and 50 mg / L tap water. After feeding for 8 months, rats were sacrificed by femoral artery excretion, and urine fluoride and bone fluoride contents of each group were measured by fluoride ion selective electrode. After HE staining, the histopathological changes and morphometry of the femur were observed under light microscope. Immunohistochemistry and in situ hybridization were used to detect the expression of SPARC protein and mRNA in osteoblasts. Results Compared with the control group [(1.36 ± 0.51) mg / L, 0.67 ± 0.16), urinary fluoride and bone fluoride contents were significantly lower in the low fluoride group [(6.09 ± 0.56) mg / L, μg / g], the difference was statistically significant (all P <0.05), the high fluorine group [(7.69 ± 0.64) mg / L, (26.53 ± 5.88) μg / g] Significance (P <0.05). Fluoride group showed the lower femur bone sclerosis. The level of SPARC protein and mRNA expression in low fluoride group (125.24 ± 1.91, 100.90 ± 1.13) was significantly higher than that in control group (109.70 ± 1.70, 88.03 ± 1.26), the difference was statistically significant (P <0.05) Group (139.47 ± 1.03,119.22 ± 1.23) lower fluoride group increased, the difference was statistically significant (P all <0.05). Conclusions Excessive fluoride regulates the proliferation, differentiation and maturation of osteoblasts by up-regulating the expression of SPARC protein and mRNA in rat osteoblasts, resulting in increased bone formation and osteosclerosis.