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目的:研究过量氟对大鼠HAT-7细胞系自噬的影响。方法:取大鼠HAT-7细胞,分别加入不同浓度的氟化钠培养液,培养48 h后,透射电子显微镜检测过量氟对大鼠成釉细胞自噬泡的影响,Western免疫印迹和定量反转录聚合酶链反应技术检测过量氟诱导大鼠成釉细胞内微管相关蛋白轻链(LC3)和Beclin 1表达的变化。选择40只Wistar大鼠,随机分为A、B、C、D 4组,进行免疫组织化学实验,检测饮水氟对大鼠自噬分子表达的影响。采用SPSS13.0软件包对数据进行统计学分析。结果:实验组自噬泡明显增多;Western免疫印迹及定量反转录聚合酶链反应结果显示,随着氟浓度的增加,HAT-7细胞自噬相关基因LC3和Beclin 1表达增加。回归分析结果显示,氟化钠与LC3及Beclin 1的表达有线性关系;大鼠体内免疫组织化学染色结果显示,实验组LC3以及Beclin 1为棕褐色,呈阳性表达。结论:过量氟诱导大鼠HAT-7细胞系自噬。
AIM: To investigate the effect of excessive fluoride on autophagy in HAT-7 cell line. Methods: Rat HAT-7 cells were treated with sodium fluoride at different concentrations for 48 h. Transmission electron microscopy was used to detect the effect of excess fluoride on autophagic vacuoles in rat ameloblasts. Western blot and quantitative Transcriptional Polymerase Chain Reaction (RT-PCR) was used to detect the expression of microtubule-associated protein light chain (LC3) and Beclin 1 in rat ameloblasts induced by excess fluoride. Forty Wistar rats were randomly divided into A, B, C and D 4 groups. Immunohistochemistry was used to detect the effect of fluoride in drinking water on the expression of autophagy in rats. SPSS13.0 software package for statistical analysis of the data. Results: The autophagy in the experimental group was significantly increased. Western blotting and quantitative reverse transcriptase-polymerase chain reaction showed that the expression of autophagy-related genes LC3 and Beclin 1 increased with the increase of fluoride concentration in HAT-7 cells. Regression analysis showed that there was a linear relationship between sodium fluoride and LC3 and Beclin 1 expression. Immunohistochemical staining results showed that LC3 and Beclin 1 in the experimental group were tan and positive. Conclusion: Excessive fluoride induces autophagy in rat HAT-7 cell line.