口腔医学 ›› 2024, Vol. 44 ›› Issue (7): 481-486.doi: 10.13591/j.cnki.kqyx.2024.07.001

• 基础与临床研究 •    下一篇

NF-κB信号通路调控对粪肠球菌脂磷壁酸刺激下巨噬细胞PD-L1表达的影响

杨艳玲1,刘润泽1,孙青1,2,范伟1,2()   

  1. 1.武汉大学口腔医学院口颌系统重建和再生全国重点实验室,口腔生物医学教育部重点实验室,口腔医学湖北省重点实验室,武汉大学口腔医(学)院,湖北武汉(430079)
    2.武汉大学口腔医院牙体牙髓二科,湖北武汉(430079)
  • 收稿日期:2024-01-03 出版日期:2024-07-28 发布日期:2024-07-15
  • 通讯作者: 范 伟 E-mail:weifan@whu.edu.cn
  • 基金资助:
    国家自然科学基金(82270968)

The effect of NF-κB signaling pathway regulation on PD-L1 expression of Enterococcus faecalis LTA-stimulated macrophages

YANG Yanling1,LIU Runze1,SUN Qing1,2,FAN Wei1,2()   

  1. State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, Wuhan 430079, China
  • Received:2024-01-03 Online:2024-07-28 Published:2024-07-15

摘要:

目的 探讨粪肠球菌脂磷壁酸(lipoteichoic acid,LTA)对巨噬细胞免疫检查点——细胞程序性死亡蛋白配体1(programmed cell death protein ligand 1,PD-L1)表达的影响及调控机制。方法 用不同浓度LTA刺激小鼠单核巨噬细胞系(RAW264.7)建立体外模型,利用流式细胞术、qRT-PCR、蛋白免疫印迹、细胞免疫荧光检测PD-L1表达与核因子κB(nuclear factor kappa-B,NF-κB)p65磷酸化水平;NF-κB通路抑制剂BAY 11-7082预处理巨噬细胞,通过以上方法进一步检测PD-L1、p65磷酸化表达变化。结果 LTA可明显诱导RAW264.7细胞中PD-L1表达,提高p65磷酸化水平(P<0.05);抑制NF-κB信号通路后,p65磷酸化水平明显降低,同时PD-L1表达减少(P<0.05)。结论 LTA可促进PD-L1在巨噬细胞中的表达,该过程受到NF-κB信号通路的调控,为进一步理解慢性根尖周炎发病机制及其治疗提供了新思路。

关键词: 粪肠球菌, 脂磷壁酸, 免疫检查点, NF-κB

Abstract:

Objective To investigate the effect of Enterococcus faecalis (E. faecalis) lipoteichoic acid (LTA) on the expression of the macrophage immune checkpoint-programmed cell death protein ligand 1 (PD-L1) and explore the possible regulatory mechanisms. Methods In vitro model was established by stimulating mouse monocyte macrophage cell line (RAW264.7) with different concentrations of E. faecalis LTA. PD-L1 expression and nuclear factor kappa-B (NF-κB) p65 phosphorylation level were detected using flow cytometry, quantitative real-time polymerase chain reaction (qRT-PCR), Western blot and cellular immunofluorescence. Macrophages were pretreated with the NF-κB signaling pathway inhibitor BAY 11-7082, then the expression of PD-L1 and p65 phosphorylation level were detected by the above methods. Results LTA induced PD-L1 expression in RAW264.7 cells, and p65 phosphorylation level increased (P<0.05). Inhibition of the NF-κB signaling pathway could reduce p65 phosphorylation level, simultaneously reducing the expression of PD-L1 on LTA-stimulated RAW264.7 cells (P<0.05). Conclusion E. faecalis LTA can promote the PD-L1 expression of macrophages and this process is regulated by the NF-κB signaling pathway, which provides further insights into the occurrence and treatment of chronic periapical inflammation.

Key words: Enterococcus faecalis, lipoteichoic acid, immune checkpoint, NF-κB

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