吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (01): 23-27.doi: 10.13481/j.1671-587x.20190105

• 基础研究 • 上一篇    

牙周炎患者自身组织核酸对小鼠巨噬细胞中NLRP3 mRNA表达的影响及其意义

袁中政1,2, 刘引1,2, 申玉芹3, 林崇韬3   

  1. 1. 吉林省牙发育及颌骨重塑与再生重点实验室, 吉林 长春 130021;
    2. 江苏省无锡口腔医院牙周科, 江苏 无锡 214000;
    3. 吉林大学口腔医院牙周科, 吉林 长春 130021
  • 收稿日期:2018-04-20 发布日期:2019-01-28
  • 通讯作者: 林崇韬,教授,硕士研究生导师(Tel:0431-88796039,E-mail:linct@jlu.edu.cn) E-mail:linct@jlu.edu.cn
  • 作者简介:袁中政(1988-),男,辽宁省沈阳市人,住院医师,医学硕士,主要从事口腔内科学方面的研究。

Effect of periodontitis patient's own tissue nucleic acid on expression of NLRP3 mRNA in murine macrophages and its significance

YUAN Zhongzheng1,2, LIU Yin1,2, SHEN Yuqin3, LIN Chongtao3   

  1. 1. Jilin Provincial Key Laboratory of Tooth Development and Bone Remodeling, Changchun 130021, China;
    2. Department of Periodontology, Wuxi Stomatology Hospital, Jiangsu Province, Wuxi 214000, China;
    3. Department of Periodontology, Stomatology Hospital, Jilin University, Changchun 130021, China
  • Received:2018-04-20 Published:2019-01-28
  • Contact: 国家自然科学基金面上项目资助课题(81371153);吉林省科技厅自然科学基金面上项目资助课题(20150101173JC) E-mail:linct@jlu.edu.cn
  • Supported by:
    BKJJ acknowledges The World Academy of Sciences (TWAS) and the International Center for Chemical and Biological Sciences (ICCBS), University of Karachi, Pakistan for their financial and technical support through the ICCBS-TWAS Postdoctoral Fellowship number 3240280476.

摘要: 目的:检测牙周炎患者自身组织核酸刺激小鼠巨噬细胞后炎性相关因子NOD样受体蛋白3(NLRP3)、半胱氨酸天冬氨酸特异蛋白酶1(caspase-1)、白细胞介素1β(IL-1β)和白细胞介素18(IL-18) mRNA表达水平,探讨牙周炎患者自身组织核酸对小鼠巨噬细胞中炎性相关因子的作用。方法:于慢性牙周炎患者翻瓣术中采集炎性牙周组织、于牙周组织健康患者冠延长术中采集健康牙周组织,分别提取组织总RNA逆转录成cDNA (cDNA质量浓度为1 mg·L-1)。培养小鼠巨噬细胞RAW264.7,分为对照组和实验组,分别加入上述提取的健康牙周组织cDNA (健康牙周组织)和炎症牙周组织cDNA (炎症牙周组织)共孵育。共孵育4、6和8h后,实时定量聚合酶链反应(RT-PCR)法检测各组RAW264.7细胞中NLRP3、caspase-1、IL-1β和IL-18 mRNA表达水平。结果:显微镜下观察,小鼠巨噬细胞RAW264.7生长状态良好,细胞形态似圆形和多边形,胞浆透明,胞体丰满。与对照组比较,实验组在4、6和8h时RAW264.7细胞中NLRP3、caspase-1、IL-1β和IL-18 mRNA表达水平明显升高(P<0.05或P<0.01),6 h时实验组RAW264.7细胞中NLRP3、caspase-1、IL-1β和IL-18 mRNA表达水平最高。结论:牙周炎患者自身组织核酸可促进小鼠巨噬细胞中炎性相关因子mRNA表达,牙周炎患者自身组织核酸对小鼠巨噬细胞的活化具有免疫调节作用。

关键词: 牙周炎, 自身组织核酸, NOD样受体蛋白3, 炎性相关因子, 巨噬细胞

Abstract: Objective: To detect the mRNA expression levels of inflammatory-related factors NLRP3, caspase-1, IL-1β, and IL-18 in the murine macrophages infected by periodontitis patient's own tissue nucleic acid, and to discuss the effects of periodontitis patient's own tissue nucleic acid on the inflammation-related factors in the macrophages.Methods: The inflammatory periodontal tissue samples were collected during periodontal flap surgery of the chronic periodontitis patients, and the healthy periodontal tissue samples were collected from the patients without any periodontal diseases undergoing crown lengthening surgery. Then the total RNA from gingival tissue was extracted and reversely transcribed into cDNA. The cultured mouse macrophages RAW264.7 were divided into control group and experiment group, then the healthy periodontal tissue cDNA and inflammatory periodontal tissue cDNA (the cDNA at a concentration of 1 mg·L-1)were added into the RAW264.7 cells,respectively. Real-time PCR was used to detect the mRNA expression levels of NLRP3, Caspase-1, IL-1β, and IL-18 in the macrophages in various groups at 4, 6 and 8 h after incubation.Results: The microscope observation showed that the mouse macrophages RAW264.7 grew well with round and polygon shapes, clear cytoplasm, and full cell body. Compared with control group, the expression levels of NLRP3, Caspase-1, IL-1β, and IL-18 mRNA in the RAW264.7 cells in experiment group at 4, 6, and 8 h were increased significantly(P<0.05 or P<0.01), and the expression levels of NLRP3,Caspase-1,IL-1β, and IL-18 mRNA in RAW264.7 cells at 6 h in experiment group were the highest.Conclusion: The periodontitis patient's own tissue nucleic acids can promote the mRNA expressions of inflammation-related factors in the RAW264.7 cells, suggesting that the periodontitis patient's own tissue nucleic acid has an immunomodulatory effect on the activation of RAW264.7 cells.

Key words: periodontitis, own tissue nucleic acid, NOD-like receptor protein 3, inflammation-related factors, macrophages

中图分类号: 

  • R781.4