Par3在成牙本质细胞胞间连接及其生物学功能中的作用研究

doi:10.13701/j.cnki.kqyxyj.2022.04.012

口腔医学研究 ›› 2022, Vol. 38 ›› Issue (4): 350-356.DOI: 10.13701/j.cnki.kqyxyj.2022.04.012

Par3在成牙本质细胞胞间连接及其生物学功能中的作用研究

王珏钰, 肖敏, 白玉, 程小刚, 吴昊泽, 况金鑫, 余擎^*

军事口腔医学国家重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔疾病重点实验室,第四军医大学口腔医院牙体牙髓病科陕西西安 710032

收稿日期:2022-01-06 出版日期:2022-04-28 发布日期:2022-04-22
通讯作者: *余擎,E-mail:yuqing@fmmu.edu.cn
作者简介:王珏钰(1996~ )女,重庆璧山人,医师,硕士在读,研究方向:细胞极性及牙本质再生。
基金资助:
国家自然科学基金(编号:81870751)陕西省自然科学基础研究计划面上项目(编号:2021JM-235)

Par3 Has an Effect on Cellular Junction and Biological Behavior of OLCs

WANG Jueyu, XAIO Min, BAI Yu, CHENG Xiaogang, WU Haoze, KUANG Jinxin, YU Qing^*

State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Oral Diseases, Department of Operative Dentistry and Endodontics, School of Stomatology, The Fourth Military Medical University, Xi'an 710032, China

Received:2022-01-06 Online:2022-04-28 Published:2022-04-22

摘要/Abstract

摘要： 目的: 通过下调分离缺陷基因3(partitioning defective-3,Par3)的表达探究其对小鼠成牙本质细胞系细胞(odontoblast lineage cells,OLCs)胞间连接的调控作用及相关生物学影响。方法: 通过免疫荧光实验分别观察Par3在大鼠牙髓组织及小鼠成牙本质细胞中的表达及分布;利用Western blot检测炎症状态下成牙本质细胞Par3的表达情况;利用小干扰RNA(siRNA)敲低成牙本质细胞Par3的表达后,通过透射电镜观察Par3表达下调前后小鼠成牙本质细胞胞间连接结构的变化;利用Western blot及细胞免疫荧光技术检测Par3表达下调前后细胞紧密连接相关分子ZO-1(zonula occludens-1)的表达及分布情况;最后,通过EdU(5-ethynyl-2’- deoxyuridine)、Transwell及流式细胞术等检测Par3表达下调对成牙本质细胞增殖、迁移及凋亡等生物学功能的影响。结果: Par3在大鼠牙髓组织中与成牙本质细胞分布一致。体外条件下Par3表达于成牙本质细胞胞质内,且在炎症状态下表达水平下降。Par3表达下调后小鼠成牙本质细胞胞间连接不连续,紧密连接相关蛋白ZO-1表达下调且其分布由细胞膜转移至细胞质;Par3表达下调组与对照组相比,细胞增殖、迁移能力增强,凋亡能力受抑制。结论: Par3表达下调可通过影响ZO-1的表达/分布破坏小鼠成牙本质细胞胞间连接,同时通过促进细胞增殖及迁移等能力增强了细胞的修复能力。

关键词: 成牙本质细胞, 细胞连接, Par3, 反应性牙本质

Abstract: Objective: To investigate the effect of partitioning defective-3 (Par3) on cell junction and biological behavior of odontoblast cells (OLCs) in vitro. Methods: The expression and distribution of Par3 in vivo and in vitro were examined by immunofluorescence (IF). Western blotting was used to assess the expression level of Par3 in OLCs at inflammatory and physiological state. Par3 siRNA was transfected in OLCs to interfere the expression of Par3. The continuity and structure of the intercellular junction were observed by transmission electronic microscopy (TEM). Western blotting and IF were performed to investigate the expression and distribution of cell junction related protein ZO-1. 5-Ethynyl-2'-deoxyuridine (EdU), transwell assay, scratch-wound assay, and flow cytometry were used to assess cell biological behavior like proliferation, migration, and apoptosis. Results: In rat pulp tissue, Par3 located along the odontoblast layer. Par3 located at cytoplasm in OLCs. The expression of Par3 might be down-regulated at inflammatory state. The cellular junction between OLCs was discontinuous and the expression of total ZO-1 or ZO-1 located at membrane in OLCs was reduced after eliminated Par3. The growth rate and the number of migrated cells in siPar3 group were increased. While the apoptosis rate was decreased in siPar3 group compared with the control group. Conclusion: Downregulation of Par3 can disrupt the intercellular junction of OLCs by regulating the expression and distribution of ZO-1, while the capacity of cellular proliferation and migration is enhanced.

Key words: odontoblast cell, cell junction, Par3, reaction dentin

王珏钰, 肖敏, 白玉, 程小刚, 吴昊泽, 况金鑫, 余擎. Par3在成牙本质细胞胞间连接及其生物学功能中的作用研究[J]. 口腔医学研究, 2022, 38(4): 350-356.

WANG Jueyu, XAIO Min, BAI Yu, CHENG Xiaogang, WU Haoze, KUANG Jinxin, YU Qing. Par3 Has an Effect on Cellular Junction and Biological Behavior of OLCs[J]. Journal of Oral Science Research, 2022, 38(4): 350-356.

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Par3在成牙本质细胞胞间连接及其生物学功能中的作用研究

Par3 Has an Effect on Cellular Junction and Biological Behavior of OLCs

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