华西口腔医学杂志 ›› 2021, Vol. 39 ›› Issue (1): 26-31.doi: 10.7518/hxkq.2021.01.004

• 基础研究 • 上一篇    下一篇

microRNA-146a对牙龈卟啉单胞菌脂多糖刺激下淋巴细胞分泌细胞因子的调节作用

司雨婷1,2(), 宋金花1,2, 方珍2, 韩晓哲3, 蒋少云1,2()   

  1. 1.天津医科大学口腔医院牙周科,天津 300070
    2.北京大学深圳医院口腔医学中心牙周科,深圳 518036
    3.美国哈佛大学牙学院Forsyth研究所,美国 剑桥 02146
  • 收稿日期:2020-02-17 修回日期:2020-10-23 出版日期:2021-02-01 发布日期:2021-03-02
  • 通讯作者: 蒋少云 E-mail:siyutingsyt401@163.com;jiangshaoyun11@126.com
  • 作者简介:司雨婷,硕士,E-mail:siyutingsyt401@163.com
  • 基金资助:
    国家自然科学基金(81670990);北京大学深圳医院院内基础研究基金(JCYJ2019004RC);美国国立口腔与颅面研究基金(R01DE025255)

MicroRNA-146a regulates the production of cytokines in lymphocytes stimulated by Porphyromonas gingivalis lipopolysaccharide

Si Yuting1,2(), Song Jinhua1,2, Fang Zhen2, Han Xiaozhe3, Jiang Shaoyun1,2()   

  1. 1.Dept. of Periodontology, Hospital of Stomatology, Tianjin Medical University, Tianjin 300070, China
    2.Dept. of Periodontology, Stomatological Center, Peking University Shenzhen Hospital, Shenzhen 518036, China
    3.The Forsyth Institute, Harvard School of Dental Medicine, Cambridge 02146, USA
  • Received:2020-02-17 Revised:2020-10-23 Online:2021-02-01 Published:2021-03-02
  • Contact: Jiang Shaoyun E-mail:siyutingsyt401@163.com;jiangshaoyun11@126.com
  • Supported by:
    The National Natural Science Foundation of China(81670990);Basic Research Grand of Peking University Shenzhen Hospital(JCYJ2019004RC);National Institute of Dental and Craniofacial Research, The United States of America(R01DE025255)

摘要: 目的

探讨microRNA-146a(miR-146a)对牙龈卟啉单胞菌(P.gingivalis)脂多糖(LPS)刺激下淋巴细胞分泌细胞因子的作用。

方法

从小鼠脾脏收集淋巴细胞。实时定量聚合酶链反应和酶联免疫反应用于检测淋巴细胞在P.gingivalis LPS、miR-146a模拟物或抑制剂处理后细胞因子的表达。

结果

P.gingivalis LPS未刺激组相比,P.gingivalis LPS能促进白细胞介素(interleukin,IL)-1β、IL-6、细胞核因子κB受体活化因子配体(RANKL)和IL-10表达(P<0.05),虽抑制骨保护素(OPG)mRNA水平(P<0.05),但分泌水平差异无统计学意义(P>0.05)。与阴性对照组相比,P.gingivalis LPS处理的淋巴细胞中miR-146a模拟物抑制IL-1β、IL-6和RANKL表达(P<0.05),促进IL-10和OPG表达(P<0.05),miR-146a抑制剂对这些细胞因子产生相反的效果(P<0.05)。

结论

miR-146a可通过抑制淋巴细胞IL-1β、IL-6和RANKL表达,促进IL-10和OPG表达,从而抑制P.gingivalis LPS的炎性作用,为骨改建提供良好的成骨环境。

关键词: 牙周炎, microRNA-146a, 细胞因子, 淋巴细胞, 脂多糖

Abstract: Objective

This study aimed to investigate the effects of microRNA-146a (miR-146a) on the production of cytokines in lymphocytes stimulated by Porphyromonas gingivalis (P.gingivalis) lipopolysaccharide (LPS).

Methods

Lymphocytes were harvested from mouse spleen and cultured in vitro. The cells were treated with P. gingivalis LPS, miR-146a mimic, or miR-146a inhibitor. Scramble RNA served as the negative control of mimic and inhibitor. The production of inflammatory cytokines was detected by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay.

Results

Compared with non-LPS-stimulated group, P. gingivalis LPS could increase the levels of interleukin (IL)-1β, IL-6, receptor activator NF-κB ligand (RANKL), and IL-10 (P<0.05) and decrease the mRNA level of osteoprotectin (OPG) (P<0.05). However, it did not significantly change the secretion of OPG. Compared with the negative control group, miR-146a mimic upregulated the levels of IL-10 and OPG (P<0.05), downregulated IL-1β, IL-6, and RANKL (P<0.05). Meanwhile, miR-146a inhibitor had a reverse effect on these cytokines (P<0.05) in P.gingivalis LPS-treated-lymphocytes.

Conclusion

MiR-146a can provide a suitable microenvironment for bone formation by preventing the inflammatory effects of P.gingivalis LPS through the inhibition of IL-1β, IL-6, and RNAKL, thereby enhancing IL-10 and OPG.

Key words: periodontitis, microRNA-146a, cytokine, lymphocytes, lipopolysaccharide

中图分类号: