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目的观察用含不同浓度氟饮水喂养的SD大鼠所产的子鼠成釉细胞中基质金属蛋白酶-20(MMP-20)、激肽释放酶(KLK4)的表达变化,探讨氟牙症的发病机制。方法给母体SD大鼠分别饮用氟质量浓度为0、50、100mg/L的饮水,待母鼠自然分娩后,取其子鼠下颌骨制作切片并提取牙胚组织,分别用免疫组化和实时定量PCR方法检测不同染氟组大鼠成釉细胞中MMP-20及KLK4的蛋白及基因表达,应用Image-Pro Plus 6.0和GraphPad Prism 5软件对免疫组化和PCR数据结果进行分析。结果①随着饮水氟质量浓度的增加,成釉细胞中MMP-20蛋白及其mRNA的表达量逐渐减少;②KLK4蛋白和KLK4mRNA的表达量在两实验组呈现了类似的变化,但高氟组与低氟组间的差别不明显。结论氟可使成釉细胞减少对MMP-20及KLK4的分泌。这可能是釉基质蛋白降解延迟、釉质矿化异常,最终导致氟牙症产生的机制之一。
Objective To observe the expression of matrix metalloproteinase-20 (MMP-20) and kallikrein (KLK4) in rat ameloblast progenitor cells from SD rats fed with different concentrations of fluoride drinking water, and to investigate the incidence of dental fluorosis mechanism. Methods The female SD rats were drank with 0, 50 and 100 mg / L fluoride respectively. After the mother gave birth naturally, the mandibular bone of the offspring was sliced and the tooth germ tissue was extracted. Immunohistochemistry and real time Quantitative PCR was used to detect the protein and gene expression of MMP-20 and KLK4 in ameloblasts of different fluoride-exposed rats. The results of immunohistochemistry and PCR were analyzed by Image-Pro Plus 6.0 and GraphPad Prism 5 software. Results ① The expression of MMP-20 protein and mRNA in ameloblasts gradually decreased with the increase of fluoride concentration in drinking water. ② The expressions of KLK4 protein and KLK4 mRNA showed similar changes in two experimental groups, The difference between low fluoride group is not obvious. Conclusion Fluoride can reduce ameloblasts secretion of MMP-20 and KLK4. This may be delayed degradation of the enamel matrix protein, enamel mineralization anomalies, and ultimately lead to one of the mechanisms of dental fluorosis.