上海交通大学学报(医学版) ›› 2020, Vol. 40 ›› Issue (4): 437-.doi: 10.3969/j.issn.1674-8115.2020.04.004

• 论著·基础研究 • 上一篇    下一篇

硫酸吲哚酚对人牙周膜细胞增殖及炎症因子表达的影响

夏欣怡,陈慧文,胡 苡,宋忠臣   

  1. 上海交通大学医学院附属第九人民医院?口腔医学院牙周病科,国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室,上海市口腔医学研究所,上海 200011
  • 出版日期:2020-04-28 发布日期:2020-05-22
  • 通讯作者: 宋忠臣,电子信箱:szhongchen@sina.com。
  • 作者简介:夏欣怡(1994—),女,住院医师,硕士生;电子信箱:765743434@qq.com。
  • 基金资助:
    国家重点研发计划(2017YFC0840100,2017YFC0840110)。

Effect of indoxyl sulfate on proliferation and of inflammatory cytokines and ROS in human periodontal ligament cells

XIA Xin-yi, CHEN Hui-wen, HU Yi, SONG Zhong-chen   

  1. Department of Periodontology, Shanghai Ninth Peoples Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai 200011, China
  • Online:2020-04-28 Published:2020-05-22
  • Supported by:
    National Key R&D Program of China (2017YFC0840100, 2017YFC0840110).

摘要: 目的·探索硫酸吲哚酚(indoxyl sulfate,IS)对人牙周膜细胞(human periodontal ligament cells,hPDLCs)增殖、炎症因子和活性氧(reactive oxygen species,ROS)表达的影响。方法·采用组织块培养法体外培养hPDLCs;采用四甲基偶氮唑盐比色(methyl thiazolyl tetrazolium,MTT)法观察IS(按0、62.5、125、250、500、1 000 μmol/L 分组)对hPDLCs增殖活性的影响;采用实时荧光定量PCR和ELISA检测各组炎症因子IL-1β、IL-6、IL-8 mRNA和蛋白的表达;采用2, 7-二氯荧光素二乙酸盐(2, 7-dichlorofluorescin diacetate,DCFDA)荧光探针检测各组hPDLCs胞内ROS的表达。结果·在24、48、72 h时,125 μmol/L的IS可抑制hPDLCs增殖,与对照组相比差异具有统计学意义(PIL-1β、IL-6、IL-8 mRNA和蛋白的表达,上调胞内ROS水平。结论·IS可通过抑制hPDLCs活性、促进炎症因子表达和提高胞内ROS水平,从而在慢性肾病与牙周炎的相互关联中起重要作用。

关键词: 牙周炎, 慢性肾衰竭, 硫酸吲哚酚

Abstract:

Objective · To explore the effects of Indoxyl sulfate (IS) on proliferation activity, of inflammatory factors and reactive oxygen species (ROS) in human periodontal ligament cells (hPDLCs). Methods · The primary hPDLCs were culturedusing tissue explant method in vitro. MTT assay was employed to evaluate the effect of IS on proliferation activity of hPDLCs. The mRNA and protein s of IL-1β, IL-6, and IL-8 were detectedusing real-time PCR and ELISA assay. DCFDA fluorescence probe was used to detect intracellular ROS and ROS in the cytoplasm under fluorescence microscope. Results · The viability of hPDLCs was inhibitedIS at the concentration of 125 μmol/L on 24, 48 and 72 hours. The inhibitory effect was presented in a dose- and time- dependent manner. IS could upregulate the mRNA and protein of inflammatory cytokines including IL-1β, IL-6 and IL-8 as well as promote ROS in hPDLCs. Conclusion · IS may play an important role in the association between chronic kidney diseases and periodontitis through inhibiting the activity of hPDLCs, promoting the of inflammatory cytokines and increasing intracellular ROS level.

Key words: periodontitis, chronic renal failure, indoxyl sulfate