NLRP6 suppresses the inflammatory response of human periodontal ligament cells by inhibiting NF-κB and ERK signal pathways
NLRP6 suppresses the inflammatory response of human periodontal ligament cells by inhibiting NF-κB and ERK signal pathways
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NLRP6通过抑制NF-κB和ERK信号通路抑制人牙周膜细胞的炎症反应
DOI：
10.1111/iej.13091
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发表时间：
2019-07-01
影响因子：
5
通讯作者：
Huang, D. M.
中科院分区：
文献类型：
--
作者：
Lu, W. L.;Zhang, L.;Huang, D. M.
目的探讨NLRP 6（NOD-，LRR- and pyrin domain-containing 6）在人牙周膜细胞（HPDLCs）炎症反应中的作用及机制。方法收集3例根尖周炎患者根管显微外科治疗后的根尖周炎组织。采用免疫组化和免疫荧光法检测NLRP 6在3例根尖周炎组织和牙周膜细胞中的表达。Western blot检测NLRP 6、Phospho（p）- p65、p65、I kappa B-α、p-I kappa B-α、ERK、p-ERK、NLRP 3、Pro interleukin（IL）-1 β、Pro caspase-1和含CARD的凋亡相关斑点样蛋白（ASC）的表达。采用定量实时聚合酶链反应和酶联免疫吸附试验检测促炎细胞因子的基因表达和分泌。采用独立样本t检验对数据进行统计学分析。结果NLRP 6在根尖周炎性组织和牙周膜细胞中均有表达。大肠杆菌脂多糖（LPS）可诱导HPDLCs中NLRP 6的表达（P < 0.05）。NLRP 6基因沉默后，E. coli LPS诱导的NF-κ B和ERK信号通路的激活增强，同时伴有IL-6和肿瘤坏死因子-α（TNF-α）水平的升高（P < 0.05）。此外，NLRP 6的敲低导致NLRP 3、Pro IL-1 β和Pro caspase-1的上调（P < 0.05），而ASC的下调（P <0.05），这可能有助于HPDLCs炎症中IL-1 β水平不变。结论NLRP 6在根尖周组织和牙周膜细胞中有功能性表达。NLRP 6通过抑制NF-κ B和ERK信号通路负性调节HPDLCs炎症中IL-6和TNF-α的产生。
Aim To explore the function and mechanisms of NLRP6 (NOD-, LRR- and pyrin domain-containing 6) in the inflammatory response of human periodontal ligament cells (HPDLCs). Methodology Tissues associated with apical periodontitis were obtained from three patients who underwent endodontic microsurgery. The expression of NLRP6 in 3 human apical periodontitis tissues and HPDLCs was examined by immunohistochemistry and immunofluorescence, respectively. The expressions of NLRP6, Phospho(p)- p65, p65, I kappa B-alpha, p- I kappa B-alpha, ERK, p- ERK, NLRP3, Pro interleukin (IL)-1 beta, Pro caspase-1 and apoptosis-associated speck-like protein containing a CARD (ASC) were examined by western blot. The gene expression and secretion of proinflammatory cytokines were detected using quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. Data were analysed statistically with independent sample t-tests. Results NLRP6 was expressed in inflammatory periapical tissues and HPDLCs. Lipopolysaccharide (LPS) from Escherichia coli induced NLRP6 in HPDLCs (P < 0.05). After silencing NLRP6, E. coli LPS-induced activation of NF-kappa B and ERK signalling was enhanced, which was also accompanied by elevated levels of IL-6 and tumour necrosis factor-alpha (TNF-alpha) (P < 0.05). Moreover, knockdown of NLRP6 led to up-regulation of NLRP3, Pro IL-1 beta and Pro caspase-1 (P < 0.05), whereas down-regulation of ASC (P < 0.05), which may contribute to unchanged levels of IL-1 beta in HPDLCs inflammation. Conclusion NLRP6 was functionally expressed in inflamed periapical tissues and HPDLCs. NLRP6 negatively regulated the production of IL-6 and TNF-alpha in HPDLCs inflammation by inhibiting NF-kappa B and ERK signal pathways.