南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (12): 1747-1754.doi: 10.12122/j.issn.1673-4254.2022.12.01

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WP1130通过抑制NLRP3炎症小体活化缓解小鼠的感染性休克

陆 莉，刘迪迪，杨燕青，王凤超   

  1. 蚌埠医学院第一附属医院检验科，安徽 蚌埠 233004；蚌埠医学院慢性疾病免疫学基础与临床安徽省重点实验室，安徽 蚌埠 233030
  • 出版日期:2022-12-20 发布日期:2023-01-12

WP1130 relieves septic shock in mice by inhibiting NLRP3 inflammasome activation

LU Li, LIU Didi, YANG Yanqing, WANG Fengchao   

  1. Department of Laboratory, First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, China; Anhui Provincial Key Laboratory of Immunology in Chronic Diseases, Bengbu Medical College, Bengbu 233030, China
  • Online:2022-12-20 Published:2023-01-12

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摘要： 目的 探究小分子化合物WP1130抑制NLRP3炎症小体活化并缓解感染性休克的作用机制。方法 细胞生物学水平：WP1130预处理小鼠骨髓来源巨噬细胞（BMDM）或人THP-1细胞后，利用多种NLRP3炎症小体活化剂（Nigericin、ATP、MSU、胞内LPS转染）活化NLRP3炎症小体，使用poly A:T活化AIM2炎症小体，通过Western blot和ELISA检测细胞培养上清中caspase-1和IL-1β的分泌水平，确定WP1130对NLRP3炎症小体的抑制效果及其特异性。利用激光共聚焦显微镜观察Nigericin诱导的线粒体损伤情况，探究WP1130是否影响NLRP3炎症小体组装活化的上游信号。动物水平：将SPF级雄性C57BL/6小鼠随机分为3组，即空白对照组（Control组）、感染性休克组（LPS组）、WP1130治疗组（WP1130+LPS组），ELISA检测小鼠血清和腹腔液中IL-1β、TNF-α的分泌水平。结果 WP1130可剂量依赖性的抑制多种激动剂诱导的NLRP3炎症小体活化（P<0.05），但对非炎症小体相关炎性因子IL-6、TNF-α的分泌无显著影响（P>0.05）。此外，WP1130对AIM2炎症小体的活化无显著影响（P>0.05）。机制研究表明，WP1130并不影响NLRP3炎症小体组装活化的上游信号线粒体损伤。动物实验结果表明，相较感染性休克组（LPS组），WP1130治疗组（WP1130+LPS组）小鼠血清和腹腔液中IL-1β的水平显著降低（P<0.05），而TNF-α的分泌水平无统计学差异（P>0.05）。结论 WP1130能够特异性抑制NLRP3炎症小体活化并缓解LPS诱导的小鼠感染性休克。

关键词: WP1130；NLRP3炎症小体；炎症小体相关疾病；感染性休克

Abstract: Objective To investigate the mechanism by which the small molecule compound WP1130 inhibits NLRP3 inflammasome activation and alleviates septic shock. Methods Mouse bone marrow-derived macrophages (BMDM) and human THP-1 cells were pre-treated with WP1130 before stimulation with different NLRP3 inflammasome agonists (Nigericin, ATP, MSU and intracellular LPS transfection), and AIM2 inflammasomes were activated with poly A: T. The levels of caspase-1 and IL-1β in the cell culture supernatant were determined using Western blotting and ELISA, and mitochondrial damage in the cells was observed using confocal microscopy. In the animal experiment, male C57BL/6 mice were randomized into blank control group, septic shock group (LPS group) and WP1130 treatment group (WP1130+LPS group), and the levels of IL-1β and TNF-α in the serum and peritoneal cavity were detected using ELISA. Results In murine BMDM and human THP-1 cells, WP1130 significantly inhibited NLRP3 agonists-induced caspase-1 and IL-1β secretion in a dose-dependent manner (P<0.05) but did not obviously affect the secretion of such inflammatory factors as IL-6 and TNF-α that were not associated with inflammasomes (P>0.05). Treatment with WP1130 did not significantly affect poly A:T-induced activation of AIM2 inflammasomes (P>0.05) or induce obvious changes in mitochondrial damage, an upstream signal of NLRP3 inflammasome activation. In the mouse model of LPS-induced septic shock, WP1130 treatment significantly reduced the level of IL-1β (P<0.05) without obviously affecting TNF-α level either in the serum or in the peritoneal cavity (P>0.05). Conclusion WP1130 specifically inhibits NLRP3 inflammasome activation to alleviate LPS-induced septic shock in mice.

Key words: WP1130; NLRP3 inflammasomes; inflammasome-driven diseases; septic shock