过量氟对小鼠髁突软骨损伤及自噬相关因子LC3、p62表达的影响

doi:10.19439/j.sjos.2024.02.001

上海口腔医学 ›› 2024, Vol. 33 ›› Issue (2): 113-116.doi: 10.19439/j.sjos.2024.02.001

过量氟对小鼠髁突软骨损伤及自噬相关因子LC3、p62表达的影响

郁莹^1,2,3, 翁清清^1,2, 罗银月^1,2, 姚姝冉^1,2, 易芳羽^1,2, 张颖^1,2

1.复旦大学附属口腔医院,上海市口腔医院,上海 200001;
2.上海市颅颌面发育与疾病重点实验室,上海 200001;
3.海军军医大学第一附属医院长海医院口腔科,上海 200433

收稿日期:2023-04-03 修回日期:2023-04-21 出版日期:2024-04-25 发布日期:2024-05-14
通讯作者: 张颖, E-mail：zhangyingcmu@vip.163.com
作者简介:郁莹（1992-）,女,研究生,E-mail：shushuandhuihui@sina.com
基金资助:
国家自然科学基金（81773369）; 上海市自然科学基金（16ZR1430600）; 上海市口腔医院创新团队项目

Damage and changes in expression of autophagy related factors LC3 and p62 of condylar cartilage in fluorosis mouse

YU Ying^1,2,3, WENG Qing-qing^1,2, LUO Yin-yue^1,2, YAO Shu-ran^1,2, YI Fang-yu^1,2, ZHANG Ying^1,2

1. Shanghai Stomatological Hospital & School of Stomatology, Fudan University. Shanghai 200001;
2. Shanghai Key Laboratory of Craniomaxillofacial Development and Diseases, Fudan University. Shanghai 200001;
3. Department of Stomatology, Changhai Hospital, the First Affiliated Hospital of Naval Medical University. Shanghai 200433, China

Received:2023-04-03 Revised:2023-04-21 Online:2024-04-25 Published:2024-05-14

摘要/Abstract

摘要： 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C57BL/6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O-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。

关键词: 慢性氟中毒, 髁突软骨, 自噬, MMP-13, LC3

Abstract: 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 received 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.

Key words: Fluorosis, Condylar cartilage, Autophagy, MMP-13, LC3

中图分类号:

R780

郁莹, 翁清清, 罗银月, 姚姝冉, 易芳羽, 张颖. 过量氟对小鼠髁突软骨损伤及自噬相关因子LC3、p62表达的影响[J]. 上海口腔医学, 2024, 33(2): 113-116.

YU Ying, WENG Qing-qing, LUO Yin-yue, YAO Shu-ran, YI Fang-yu, ZHANG Ying. Damage and changes in expression of autophagy related factors LC3 and p62 of condylar cartilage in fluorosis mouse[J]. Shanghai Journal of Stomatology, 2024, 33(2): 113-116.

参考文献

[1] 张素洁, 张忠诚, 徐祗云. 微量元素氟与人体健康[J]. 微量元素与健康研究, 2007, 24(2): 69-70.
[2] 胡德渝. 口腔预防医学[M]. 北京: 人民卫生出版社, 2012: 97-98.
[3] Nishida T, Kubota S, Aoyama E, et al.Impaired glycolytic metabolism causes chondrocyte hypertrophy-like changes via promotion of phospho-Smad1/5/8 translocation into nucleus[J]. Osteoarthritis Cartilage, 2013, 21(5): 700-709.
[4] Zhao Y, Hao J, Wang J, et al.Effect of choline on the composition and degradation enzyme of extracellular matrix of mice chondrocytes exposed to fluoride[J]. Biol Trace Elem Res, 2017, 175(2): 414-420.
[5] 郑冬冬. ClC-7参与氟斑牙和氟骨症发生的实验动物研究[D]. 西安: 第四军医大学, 2013.
[6] Chen S, He LH, Zhao L, et al.Effects of articular disc or condylar cartilage resection on mandibular growth in young rats[J]. Arch Oral Biol, 2019, 97: 67-71.
[7] 安康, 张元宇, 魏亚男, 等. DON毒素与氟对大鼠骨及关节软骨的损伤作用[J]. 中国地方病防治杂志, 2016, 31(11): 1201-1204.
[8] 李辉, 黄林剑, 谢千阳, 等. 体外培养的兔下颌骨髁突对机械压力的反应和变化[J]. 中国口腔颌面外科杂志, 2022, 20(4): 340-346.
[9] 许鹏, 郭雄, 姚建锋, 等. 过量氟对实验大鼠软骨细胞分化及Ⅱ和Ⅹ型胶原表型表达的影响[J].中华风湿病学杂志, 2002, 6(4): 231-234.
[10] Hu Q, Ecker M.Overview of MMP-13 as a promising target for the treatment of osteoarthritis[J]. Int J Mol Sci, 2021, 22(4): 1742-1762.
[11] 马瑞. 氟通过PHD2/HIF1α信号通路抑制SW1353软骨细胞糖酵解[D]. 沈阳:中国医科大学, 2021.
[12] 陈荣. 自噬相关因子Beclin1、LC3和p62在慢性氟中毒大鼠软骨中的表达意[D]. 贵阳: 贵州医科大学, 2017.
[13] Li JX, Yan Q, Liu N, et al.The prognostic value of autophagy-related markers Bclin-1 and LC-3 in colorectal cancers: a systematic review and meta-analysis[J]. Evid Based Complement Alternat Med, 2020: 8475840.
[14] 隋馨莹, 徐平, 段昌柱,等. p62蛋白的分子功能及其在疾病中的研究进展[J]. 生物工程学报, 2023, 39(4): 1374-1389.
[15] 翁清清, 易芳羽, 郁莹, 等. 自噬在氟牙症大鼠成釉细胞中的保护作用[J]. 口腔医学, 2020, 40(11): 982-986.

过量氟对小鼠髁突软骨损伤及自噬相关因子LC3、p62表达的影响

Damage and changes in expression of autophagy related factors LC3 and p62 of condylar cartilage in fluorosis mouse

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