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目的观察Wnt/β-catenin信号通路及其相关信号分子蛋白及mRNA表达水平在实验性氟中毒大鼠肾组织中的变化,探讨氟中毒发病机制。方法选择健康成年SD大鼠36只,雌雄各半,随机分为:对照组(饮水含氟量<1 mg/L),低、高氟组(饮水加氟量5、50 mg/L),每组12只,观察大鼠氟斑牙发生情况,测定尿氟、骨氟含量及肾功能变化;光镜观察肾组织病理形态学变化;采用实时荧光定量PCR和免疫组化方法检测肾组织中Wnt4、β-连环蛋白(β-catenin)、E-粘钙蛋白(E-cadherin)蛋白及mRNA表达。结果低氟组大鼠氟斑牙检出率为75%(9/12),高氟组为91.7%(11/12),与对照组比较,染氟组大鼠尿氟、骨氟含量随染氟浓度增加而逐渐升高,差异有统计学意义(P<0.05);染氟组大鼠血清肌酐及尿素氮明显高于对照组(均P<0.05);光镜观察染氟组大鼠肾小管上皮细胞明显水肿、渗出、坏死,细胞间界限不清,且随着染氟浓度增高,损伤加重;低、高氟组大鼠肾组织中Wnt4、β-catenin、E-cadherin蛋白表达分别为[(7.78±1.17)、(6.66±0.59)、(3.23±0.82)]和[(17.46±3.18)、(12.46±0.89)、(1.23±0.35)],mRNA表达分别为[(3.01±0.81)、(1.86±0.43)、(0.276±0.12)]和[(5.79±0.86)、(5.84±1.19)、(0.12±0.46)],与对照组比较,氟中毒组大鼠肾组织中Wnt4、β-catenin蛋白和mRNA表达升高,而E-cadherin蛋白及mRNA表达降低。结论慢性氟中毒时,氟通过刺激Wnt信号通路中Wnt4、β-catenin过表达、E-cadherin低表达,导致肾小管上皮细胞间充质转分化(EMT)。
Objective To observe the changes of Wnt / β-catenin signaling pathway and its related signal protein and mRNA expression in renal tissue of rats with experimental fluorosis, and to explore the pathogenesis of fluorosis. Methods Thirty-six healthy adult Sprague-Dawley rats were randomly divided into control group (drinking water fluoride content <1 mg / L), low and high fluoride group (water and fluoride content 5, 50 mg / L) Twelve rats in each group were used to observe the occurrence of dental fluorosis in rats. Urinary fluorine and bone fluoride contents and changes of renal function were observed. Morphological changes of renal tissues were observed with light microscopy. Real-time fluorescence quantitative PCR and immunohistochemistry Wnt4, β-catenin, E-cadherin protein and mRNA expression. Results The detection rate of dental fluorosis in low fluoride group was 75% (9/12) and that in high fluoride group (91.7%, 11/12). Compared with the control group, (P <0.05). Serum creatinine and urea nitrogen were significantly higher in the fluoride-exposed group than those in the control group (all P <0.05) The renal tubular epithelial cells were significantly edema, exudation, necrosis, unclear cell boundaries, and with the increase of fluoride concentration, the injury was aggravated. The protein expressions of Wnt4, β-catenin and E-cadherin Were (7.78 ± 1.17), (6.66 ± 0.59), (3.23 ± 0.82) and (17.46 ± 3.18), (12.46 ± 0.89) and (1.23 ± 0.35) 0.81), (1.86 ± 0.43), (0.276 ± 0.12) and (5.79 ± 0.86), (5.84 ± 1.19) and (0.12 ± 0.46) respectively. Compared with the control group, Wnt4 , Β-catenin protein and mRNA expression increased, while E-cadherin protein and mRNA expression decreased. Conclusion Fluoride can induce tubular epithelial mesenchymal transdifferentiation (EMT) through the overexpression of Wnt4, β-catenin and low expression of E-cadherin in Wnt signaling pathway during chronic fluorosis.