NLRP3炎症小体抑制剂MCC950通过抑制Th1/Th17和促进Tregs改善脓毒症小鼠器官损伤

doi:10.3969/j.issn.2095-9400.2024.03.003

科研成果详情

题名	NLRP3炎症小体抑制剂MCC950通过抑制Th1/Th17和促进Tregs改善脓毒症小鼠器官损伤
其他题名	NLRP3 inflammasome inhibitor MCC950 ameliorates sepsis-associated organ damage via suppressing Th1/Th17 response and promoting Tregs response in mice
作者	叶玉妹 1; 陈隆望 2; 张万里 3; 王雪涛 4
发表日期	2024-03-25
发表期刊	温州医科大学学报影响因子和分区
语种	中文
原始文献类型	学术期刊
关键词	MCC950 NLRP3 脓毒症 T细胞分化器官损伤
其他关键词	MCC950 ; NLRP3 ; sepsis ; T cell differentiation ; organ damage
摘要	目的：探讨核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体抑制剂MCC950对脓毒症小鼠T细胞分化及器官损伤的影响。方法：将18只C57BL/6雄性小鼠随机分为假手术组（sham组）、脓毒症模型组（CLP组）和MCC950治疗组（MCC950组），每组6只。其中CLP组和MCC950组通过盲肠结扎穿孔（CLP）的方法制备脓毒症模型。MCC950组腹腔注射溶于0.5 mL 0.9%氯化钠溶液的MCC950(50 mg/kg),sham组和CLP组腹腔注射等体积的0.9%氯化钠溶液。CLP术后24 h收集小鼠血清及器官标本。HE染色法观察脓毒症小鼠肺脏、肾脏组织病理损伤程度，并计算生存率；Western blot法检测脾脏NLRP3蛋白含量；ELISA法检测小鼠血清和肺组织的细胞因子水平（IL-1β、IL-18、IL-1α、IL-6、TNF-α）；分离脾脏获得脾脏淋巴细胞，采用流式细胞术检测辅助性T细胞（Th）1、Th17、调节性T细胞（Tregs）比例。结果：与sham组相比，CLP组小鼠肺组织炎性细胞浸润、水肿和肺泡壁增厚，肾小管刷状缘和上皮细胞减少，肾小球萎缩，肾小管成弥漫性扩张，大量炎性细胞浸润，脾脏NLRP3的蛋白表达上调（P<0.05），血清IL-1β、IL-18、IL-1α、IL-6、TNF-α水平均升高（P<0.05），肺组织中IL-1α、IL-6、TNF-α水平均升高（P<0.05），脾脏CD4+IFN-γ+Th1、CD4+IL-17+Th17、CD4+CD25+Foxp3+Tregs比例均升高（P<0.05），小鼠7 d生存率为30%(P<0.05)。与CLP组相比，MCC950组小鼠肺组织炎性细胞浸润减轻，水肿减退，肾小管扩张不明显，炎性细胞浸润亦有所减轻，脾脏NLRP3的蛋白表达下调（P<0.05），血清IL-1β、IL-18、IL-1α、IL-6、TNF-α水平均降低（P<0.05），肺组织中IL-1α、IL-6、TNF-α水平均降低（P<0.05），脾脏CD4+IFN-γ+Th1、CD4+IL-17+Th17比例减少，CD4+CD25+Foxp3+Tregs比例明显增加（P<0.05），小鼠7 d生存率显著提升（P<0.05）。结论：MCC950阻断NLRP3可改善脓毒症小鼠的器官损伤，其作用可能依赖于抑制Th1/Th17反应和促进Tregs反应。
其他摘要	Objective:To investigate the effects of MCC950,the NLRP3 inflammasome inhibitor,on T cell differentiation and organ damage in septic mice.Methods:Eighteen C57BL/6 male mice were randomly divided into sham group,CLP group and MCC950 group,with six in each.In CLP group and MCC950 group,the sepsis model was operated by cecal ligation and puncture(CLP).MCC950 group was injected intraperitoneally with MCC950(50 mg/kg)dissolved in 0.5 mL saline,and the sham group and CLP group were injected with an equal volume of saline.Mouse serum and issue were collected at 24 h after surgery.HE staining was used to observe the histopathological damage in lungs and kidneys.The survival rate was calculated in 7 d after operation;The level of splenic NLRP3 protein was detected by Western blot;IL-1β,IL-18,IL-1α,IL-6,and TNF-α protein level in the serum and lung was measured by ELISA;The proportion of helper T cells(Th)1,Th17,and regulatory T cells(Tregs)in splenic lymphocytes were detected by flow cytometry.Results:Compared with the sham group,the septic mice had inflammatory cell infiltration,oedema and alveolar wall thickening in lungs(P<0.05),and reduction of renal tubular brush border and epithelial cells,glomerular atrophy,diffusely dilatation of the renal tubules with many inflammatory cell infiltration(P<0.05).The expression of NLRP3 in the spleen and the levels of IL-1β,IL-18,IL-1α,IL-6,and TNF-α in the serum and lung in CLP group were higher than that in sham group(P<0.05).Furthermore,compared with sham group,the proportions of CD4+IFN-γ+ Th1 and CD4+IL-17+ Th17 in the spleen were elevated,accompanied by a slight increase in the proportion of CD4+CD25+Foxp3+ Tregs(P<0.05).The 7 d survival rate of septic mice was 30%.Meanwhile,the inflammatory cell infiltration and oedema in the lung in septic mice were attenuated by MCC950 treatment(P<0.05).The glomerular telangiectasia was not obvious,and the dilatation of the renal tubules was decreased in the MCC950 group(P<0.05).Compared with CLP group,the expression of NLRP3 in the spleen in CLP group was down-regulated,and the levels of IL-1β,IL-18,IL-1α,IL-6,and TNF-α in the serum and lung were reduced(P<0.05).In addition,the proportion of CD4+IFN-γ+ Th1 and CD4+IL-17+ Th17 was decreased,and the proportion of CD4+CD25+Foxp3+ Tregs was more significantly increased than that in CLP group(P<0.05).The 7 d survival rate of septic mice was improved(P<0.05).Conclusion:Inhibition of NLRP3 activation by MCC950 ameliorated organ damage in septic mice,and its effect may depend on inhibiting Th1/Th17 response and promoting Tregs response.
资助项目	温州市基础性科研项目（Y20210095）;
ISSN	2095-9400
卷号	54 期号:03 页码:190-198
DOI	10.3969/j.issn.2095-9400.2024.03.003
页数	9
收录类别	CNKI ; 万方 ; 维普
URL	查看原文
引用统计
文献类型	期刊论文
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/211231
专题	附属第一医院_超声影像科附属第一医院附属第一医院_急诊科
通讯作者	王雪涛
作者单位	1.温州医科大学附属第一医院超声影像科; 2.温州医科大学附属第一医院急诊科; 3.温州市龙湾区第一人民医院急诊科; 4.温州市龙湾区第一人民医院重症医学科
第一作者单位	超声影像科; 附属第一医院
第一作者的第一单位	超声影像科
推荐引用方式 GB/T 7714	叶玉妹,陈隆望,张万里,等. NLRP3炎症小体抑制剂MCC950通过抑制Th1/Th17和促进Tregs改善脓毒症小鼠器官损伤[J]. 温州医科大学学报,2024,54(03):190-198.
APA	叶玉妹, 陈隆望, 张万里, & 王雪涛. (2024). NLRP3炎症小体抑制剂MCC950通过抑制Th1/Th17和促进Tregs改善脓毒症小鼠器官损伤. 温州医科大学学报, 54(03), 190-198.
MLA	叶玉妹,et al."NLRP3炎症小体抑制剂MCC950通过抑制Th1/Th17和促进Tregs改善脓毒症小鼠器官损伤".温州医科大学学报 54.03(2024):190-198.