W L Lu1,2, L Zhang1,3, D Z Song1,3, X W Yi1,3, W Z Xu1,3, L Ye1,3, D M Huang1,3. 1. State Key Laboratory of Oral Diseases and National Clinical Research Center for Oral Diseases, Chengdu, China. 2. Department of Endodontics, Stomatological Hospital of Chongqing Medical University, Chongqing, China. 3. Department of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, Chengdu, China.
Abstract
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, IκB-α, p- IκB-α, ERK, p- ERK, NLRP3, Pro interleukin (IL)-1β, 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-κB and ERK signalling was enhanced, which was also accompanied by elevated levels of IL-6 and tumour necrosis factor-α (TNF-α) (P < 0.05). Moreover, knockdown of NLRP6 led to up-regulation of NLRP3, Pro IL-1β and Pro caspase-1 (P < 0.05), whereas down-regulation of ASC (P < 0.05), which may contribute to unchanged levels of IL-1β in HPDLCs inflammation. CONCLUSION: NLRP6 was functionally expressed in inflamed periapical tissues and HPDLCs. NLRP6 negatively regulated the production of IL-6 and TNF-α in HPDLCs inflammation by inhibiting NF-κB and ERK signal pathways.
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, IκB-α, p- IκB-α, ERK, p- ERK, NLRP3, Pro interleukin (IL)-1β, 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-κB and ERK signalling was enhanced, which was also accompanied by elevated levels of IL-6 and tumour necrosis factor-α (TNF-α) (P < 0.05). Moreover, knockdown of NLRP6 led to up-regulation of NLRP3, Pro IL-1β and Pro caspase-1 (P < 0.05), whereas down-regulation of ASC (P < 0.05), which may contribute to unchanged levels of IL-1β in HPDLCs inflammation. CONCLUSION:NLRP6 was functionally expressed in inflamed periapical tissues and HPDLCs. NLRP6 negatively regulated the production of IL-6 and TNF-α in HPDLCs inflammation by inhibiting NF-κB and ERK signal pathways.