高碱抗性粪肠球菌通过ZBP1诱导弱化的巨噬细胞泛凋亡

doi:10.13591/j.cnki.kqyx.2026.04.004

摘要/Abstract

摘要：

目的探讨高碱抗性粪肠球菌(highly alkali-resistant Enterococcus faecalis,HAR E. faecalis)如何调控Z-DNA结合蛋白1(Z-DNA binding protein 1,ZBP1)以及巨噬细胞的泛凋亡(焦亡、凋亡、坏死)。方法通过连续碱性传代培养构建高碱抗性粪肠球菌。使用高碱抗性及标准粪肠球菌(Enterococcus faecalis,E. faecalis)感染RAW264.7巨噬细胞,采用流式细胞术、活/死细胞染色、共聚焦显微镜成像分析和蛋白免疫印迹法,分析巨噬细胞ZBP1的表达水平以及死亡途径。使用小干扰RNA敲低巨噬细胞ZBP1的表达后进行相同感染,采用蛋白免疫印迹法分析敲低ZBP1后巨噬细胞泛凋亡标志物表达的变化。结果高碱抗性粪肠球菌上调巨噬细胞中ZBP1和泛凋亡标志物表达的能力较标准粪肠球菌弱。敲低ZBP1后,上述指标的表达均下降,且菌株间无显著差异。结论高碱抗性粪肠球菌可能通过微弱激活ZBP1,诱导弱化的巨噬细胞泛凋亡,削弱了机体彻底清除细菌的能力,从而有利于持续性根尖周炎的发生与发展。

关键词: 粪肠球菌, 碱抗性, Z-DNA结合蛋白1, 泛凋亡

Abstract:

Objective To investigate how highly alkali-resistant Enterococcus faecalis(HAR E. faecalis) regulates Z-DNA binding protein 1(ZBP1) and macrophage PANoptosis(pyroptosis, apoptosis, necrosis). Methods Highly alkali-resistant Enterococcus faecalis was constructed by continuous alkaline subculturing. RAW264.7 macrophages were infected with either the highly alkali-resistant or standard Enterococcus faecalis(E. faecalis). Flow cytometry, live/dead cell staining, confocal microscopy, and Western blot were employed to analyze ZBP1 expression levels and macrophage death pathways. Small interfering RNA(siRNA) was used to knock down ZBP1 expression in macrophages, followed by the same infection protocol. Western blot was used to analyze changes in the expression of PANoptosis markers after ZBP1 knockdown. Results The highly alkali-resistant Enterococcus faecalis exhibited a weaker ability to upregulate the expression of ZBP1 and PANoptosis markers in macrophages compared to the standard strain. Following ZBP1 knockdown, the expression of these markers decreased, with no significant differences observed between the two strains. Conclusion Highly alkali-resistant Enterococcus faecalis induces compromised macrophage PANoptosis by weak activation of ZBP1, which impairs the host’s ability to completely eradicate the bacteria, thereby possibly favoring the occurrence and development of persistent apical periodontitis.

Key words: Enterococcus faecalis, alkali-resistance, Z-DNA binding protein 1, PANoptosis

中图分类号:

R781.3

肖亦方, 刘润泽, 欧东晨, 范伟. 高碱抗性粪肠球菌通过ZBP1诱导弱化的巨噬细胞泛凋亡[J]. 口腔医学, 2026, 46(4): 260-265.

XIAO Yifang, LIU Runze, OU Dongchen, FAN Wei. Highly alkali-resistant Enterococcus faecalis induces compromised PANoptosis of macrophage via ZBP1[J]. Stomatology, 2026, 46(4): 260-265.

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基因	引物序列(5'—3')
siZBP1	F:GGACAUAGAAAGCUCUCAATT R:UUGAGAGCUUUCUAUGUCCTT
NC-siRNA	F:UUCUCCGAACGUGUCACGUTT R:ACGUAGCACGUUCGGAGAATT