Lu L, Liu D, Yang Y, Wang F
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.
Nan Fang Yi Ke Da Xue Xue Bao. 2022 Dec 20;42(12):1747-1754. doi: 10.12122/j.issn.1673-4254.2022.12.01.
To investigate the mechanism by which the small molecule compound WP1130 inhibits NLRP3 inflammasome activation and alleviates septic shock.
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.
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 ( < 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 (>0.05). Treatment with WP1130 did not significantly affect poly A: T-induced activation of AIM2 inflammasomes (>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 ( < 0.05) without obviously affecting TNF-α level either in the serum or in the peritoneal cavity (>0.05).
WP1130 specifically inhibits NLRP3 inflammasome activation to alleviate LPS-induced septic shock in mice.
研究小分子化合物WP1130抑制NLRP3炎性小体激活并减轻脓毒症休克的机制。
用不同的NLRP3炎性小体激动剂（尼日利亚菌素、ATP、MSU和细胞内脂多糖转染）刺激小鼠骨髓来源的巨噬细胞（BMDM）和人THP-1细胞之前，先用WP1130进行预处理，并用聚腺苷酸:胸腺嘧啶核苷酸激活AIM2炎性小体。采用蛋白质免疫印迹法和酶联免疫吸附测定法测定细胞培养上清液中半胱天冬酶-1和白细胞介素-1β的水平，并用共聚焦显微镜观察细胞中的线粒体损伤。在动物实验中，将雄性C57BL/6小鼠随机分为空白对照组、脓毒症休克组（脂多糖组）和WP1130治疗组（WP1130+脂多糖组），采用酶联免疫吸附测定法检测血清和腹腔中白细胞介素-1β和肿瘤坏死因子-α的水平。
在小鼠BMDM和人THP-1细胞中，WP1130以剂量依赖的方式显著抑制NLRP3激动剂诱导的半胱天冬酶-1和白细胞介素-1β分泌（<0.05），但对与炎性小体无关的白细胞介素-6和肿瘤坏死因子-α等炎症因子的分泌没有明显影响（>0.05）。WP1130处理对聚腺苷酸:胸腺嘧啶核苷酸诱导的AIM2炎性小体激活没有显著影响（>0.05），也未引起NLRP3炎性小体激活的上游信号线粒体损伤的明显变化。在脂多糖诱导的脓毒症休克小鼠模型中，WP1130治疗显著降低了白细胞介素-1β水平（<0.05），对血清或腹腔中的肿瘤坏死因子-α水平没有明显影响（>0.05）。
WP1130特异性抑制NLRP3炎性小体激活，以减轻脂多糖诱导的小鼠脓毒症休克。
