Damage and changes in expression of autophagy related factors LC3 and p62 of condylar cartilage in fluorosis mouse
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目的:研究过量氟对小鼠髁突软骨损伤及髁突软骨中自噬相关因子的表达.方法:将30只4周龄雄性C57BU6小鼠随机分为对照组和实验组,每组15只,对照组自由饮用去离子水,实验组自由饮用含氟离子浓度为75 mg/L的去离子水,持续喂养8周.通过改良番红0-固绿软骨染色观察髁突纤维软骨层结构变化;免疫组织化学染色检测软骨细胞外基质降解标志物基质金属蛋白酶13(Matrix metalloproteinase 13,MMP-13)、Ⅱ型胶原蛋白的表达水平及自噬相关因子微管相关蛋白轻链3(microtubule-associated protein light chain 3,LC3)、自噬底物蛋白p62的表达水平.采用SPSS 22.0软件包对免疫组织化学染色半定量结果进行双因素方差分析.结果:与对照组相比,实验组纤维软骨层变薄,软骨细胞体积变小,核变小,染色加深.免疫组织化学染色结果显示,实验组MMP-13表达量增加,Ⅱ型胶原蛋白表达量降低;LC3表达量增加,p62表达量下降.结论:用含氟离子浓度为75 mg/L的去离子水喂养小鼠8周,可导致髁突软骨退变,髁突软骨发生自噬.
PURPOSE:To study the damage and the expression of LC3 and p62 of condylar cartilage in fluorosis mouse.METHODS:Thirty 4-week-old male C57BL/6 mice were randomly divided into control group and the experimental group with 15 animals in each group.The control group received regular drinking water and the experimental group re-ceived a fluoride concentration of 75 mg/L drinking water for 8 weeks.The structure of condylar cartilage was observed through modified safranine O-fast green FCF cartilage stain kit.Immunohistochemistry was used to detect the expression of MMP-13,type Ⅱ collagen and LC3 and p62.Two-way analysis of variance test was conducted for analysis of semi-quantitative results of immunohistochemistry using SPSS 22.0 software package.RESULTS:Compared with the control group,the fibrocartilage layer of the experimental group became thinner,the condrocytes were smaller,and the staining became deeper.Immunohistochemistry results showed that the expression of MMP-13 and LC3 increased;the expression of type Ⅱ collagen and p62 decreased in the experimental group.CONCLUSIONS:There was degeneration of the condylar cartilage and autophagy in mice with drinking water containing 75 mg/L fluoride.
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