摘要
目的探究Avitinib抑制NLRP3炎症小体活化并缓解感染性休克的作用及其机制。方法预先给药Avitinib作用小鼠骨髓来源巨噬细胞(BMDM)、人单核细胞白血病细胞系(THP-1)、健康志愿者外周血分离提取的单个核细胞(PBMC),后加入多种NLRP3炎症小体激动剂如尼日利亚菌素(Nigericin),单钠尿酸盐(MSU)结晶,三磷酸腺苷(ATP)活化经典NLRP3炎症小体,或在细胞内转染脂多糖(LPS)活化非经典形式NLRP3炎症小体(Western blotting)检测NLRP3炎症小体活化的分泌蛋白指征及细胞焦亡情况。利用酶联免疫吸附技术(ELISA)测定细胞上清中相关炎症因子的水平。构建小鼠感染性休克模型,采取随机分组的方法,将8周龄雄性C57BL/6J小鼠分为空白对照组(Control组),感染性休克模型组(LPS组),给药组(LPS+Avitinib组),6只/组。ELISA测定各组眼球血和腹腔灌洗液中的相关炎症因子水平,并记录小鼠的存活情况,采用Kaplan-Meier法进行生存分析。结果Avitinib在多种细胞类型中抑制NLRP3炎症小体活化,剂量依赖性地抑制IL-1β分泌和caspase-1剪切,同时抑制GSDMD介导的细胞焦亡(P<0.05),但对炎症早期预警因子白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)的分泌并无影响(P>0.05)。动物实验结果表明,在LPS诱导的感染性休克模型中,给予Avitinib干预后小鼠血清和腹腔液中IL-1β的水平降低(P<0.05),并延长小鼠生存时间(P<0.05)。结论Avitinib可抑制NLRP3炎症小体活化并改善感染性休克。
Objective To investigate the effect of avitinib for suppressing NLRP3 inflammasome activation and alleviating septic shock and explore the underlying mechanism.Methods Mouse bone marrow-derived macrophages(BMDM),human monocytic leukemia cell line THP-1,and peripheral blood mononuclear cells(PBMC)isolated from healthy volunteers were pre-treated with avitinib,followed by activation of the canonical NLRP3 inflammasome using agonists including nigericin,monosodium urate(MSU)crystals,or adenosine triphosphate(ATP).Non-canonical NLRP3 inflammasome activation was induced via intracellular transfection of lipopolysaccharide(LPS).Western blotting was used to detect the secretory protein markers of NLRP3 inflammasome activation and assess pyroptosis,and the levels of inflammatory cytokines in cell culture supernatant were determined with ELISA.In a mouse model of LPS-induced septic shock,the effect of avitinib treatment on the levels of inflammatory cytokines in serum and peritoneal lavage fluid were examined with ELISA,and survival curves of the mice were plotted using the Kaplan-Meier method.Results Avitinib significantly inhibited NLRP3 inflammasome activation in multiple cell types,and dose-dependently reduced IL-1βsecretion and caspase-1 cleavage while suppressing GSDMD-mediated pyroptosis without obviously affecting IL-6 or TNF-αlevels.In the mouse models of LPS-induced septic shock,avitinib significantly lowered IL-1βlevels in serum and peritoneal fluid and extended survival time of the mice.Conclusion Avitinib suppresses NLRP3 inflammasome activation and alleviates septic shock in mice.
作者
尚菲菲
师晓可
曾尧
陶循浅
李天真
梁艳
杨燕青
宋传旺
SHANG Feifei;SHI Xiaoke;ZENG Yao;TAO Xunqian;LI Tianzhen;LIANG Yan;YANG Yanqin;SONG Chuanwang(Anhui Provincial Key Laboratory of Immunology in Chronic Diseases,Bengbu Medical University,Bengbu 233030,China;Clincial Laboratory,First Affiliated Hospital of Bengbu Medical University,Bengbu 233004,China)
出处
《南方医科大学学报》
北大核心
2025年第8期1697-1705,共9页
Journal of Southern Medical University
基金
安徽省科研编制计划优秀青年科研项目(2022AH030140)
蚌埠医科大学2024年度研究生科研创新计划项目(Byycx24010,Byycx24030)
2025年大学生创新创业训练计划项目
慢性病免疫学基础与临床安徽省重点实验室开放课题基金(AHIAI2022K01)。
