NLRP3炎症小体介导的巨噬细胞极化在ITP中的作用  

NLRP3 inflammasome mediated macrophage polarization in ITP

作　　者：宋传龙 海力其古丽·努日丁[1] 焦红杰 王学梅[1] 严媚[1] Song Chuanlong;Hailiqiguli·Nuridin;Jiao Hongjie;Wang Xuemei;Yan Mei(Dept of Pediatrics,The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054)

机构地区：[1]新疆医科大学第一附属医院儿内一科,乌鲁木齐830054

出　　处：《安徽医科大学学报》2023年第1期132-139,共8页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82160031)。

摘　　要：目的探讨在免疫性血小板减少症(ITP)中NLRP3炎症小体的活化水平及抑制NLRP3介导的炎症小体活化对M1型巨噬细胞极化与免疫功能的影响。方法RT-qPCR法检测ITP患者(ITP组)和健康对照组(Control组)外周血单个核细胞(PBMC)与CD14+单核细胞中NLRP3 mRNA的表达;ELISA法检测两组血清中IL-1β与IL-18的含量;Pearson相关系数分析NLRP3、IL-1β与IL-18表达水平与血小板计数的相关性;将ITP患者来源的M0型巨噬细胞(MDMs)分为4组:IgG对照组(IgG组),MCC950处理组(MCC950组),LPS、IFN-γ与IgG处理组(LPS+IFN-γ+IgG组)和LPS、IFN-γ与MCC950处理组(LPS+IFN-γ+MCC950组);RT-qPCR与Western blot检测4组MDMs中M1型巨噬细胞标志物CD86、iNOS、MCP-1 mRNA与蛋白水平;Western blot检测各组MDMs中NLRP3炎症小体相关蛋白NLRP3、ASC、cleaved caspase-1与IL-β的表达;流式细胞术检测各组MDMs对血小板的吞噬能力,CFSE法检测各组MDMs对CD4^(+)T与CD8^(+)T的增殖影响。结果与Control组相比,ITP组NLRP3 mRNA水平和IL-1β与IL-18含量均升高(P<0.05);ITP患者中血小板计数与NLRP3、IL-1β和IL-18呈负相关关系(P<0.05)。与IgG组相比,LPS+IFN-γ+IgG组与LPS+IFN-γ+MCC950组细胞中CD86、iNOS、MCP-1的mRNA与蛋白表达水平增加(P<0.05),NLRP3、ASC、cleaved caspase-1、IL-β蛋白表达与血小板吞噬能力和对CD4^(+)T与CD8^(+)T细胞的增殖促进能力升高(P<0.05),而与LPS+IFN-γ+IgG组相比,LPS+IFN-γ+MCC950组细胞中上述检测指标均降低(P<0.05)。结论在ITP中NLRP3炎症小体活化水平升高,且与MDMs的过度M1型极化相关,而抑制NLRP3介导的炎症小体活化可改善巨噬细胞的M1型极化与免疫功能。Objective To investigate the activation level of NLRP3 inflammasome in immune thrombocytopenia(ITP)and the effect of inhibiting NLRP3 mediated inflammasome activation on polarization and immune function of M1 macrophages.Methods The expression of NLRP3 mRNA in peripheral blood mononuclear cells(PBMC)and CD14+monocytes of ITP patients(ITP group)and Control group(Control group)was detected by RT-qPCR.The levels of IL-1βand IL-18 in serum of the two groups were determined by ELISA.M0 macrophages(MDMs)from ITP group were divided into 4 groups:IgG control group(IgG group),MCC950 treatment group(MCC950 group),LPS,IFN-γand IgG treatment group(LPS+IFN-γ+IgG group)and LPS,IFN-γand MCC950 treatment group(LPS+IFN-γ+MCC950 group);mRNA and protein levels of M1 macrophage markers CD86,iNOS and MCP-1 were detected by RT-qPCR and Western blot.Western blot was used to detect the expression of NLRP3 inflammasome associated protein,ASC,Cleaved caspase-1 and IL-β.Flow cytometry was used to detect the phagocytosis of MDMs on platelets in each group,and CFSE was used to detect the proliferation of CD4^(+)T and CD8^(+)T.Results Compared with the control group,the expression of NLRP3 mRNA in PBMC and CD14+monocytes,and the concentration of IL-1βand IL-18 in serum of ITP group increased significantly(P<0.05).Platelet counts were negative correlated with NLRP3 mRNA expression in CD14+monocyte and the concentration of IL-1β,IL-18 in serum in patients with ITP(P<0.05).Compared with IgG group,the mRNA and protein expressions of M1 macrophage markers CD86,iNOS,MCP-1,and the protein expression level of NLRP3,ASC,cleaved caspase-1 and IL-β,the platelet phagocytosis and the proliferation promoting ability of CD4^(+)T and CD8^(+)T cells significantly increased in LPS+IFN-γ+IgG group and LPS+IFN-γ+MCC950 group(all P<0.05).Compared with LPS+IFN-γ+IgG group,the above indexes significantly decreased in LPS+IFN-γ+MCC950 group(P<0.05).Conclusion The activation level of NLRP3 inflammasome in ITP is abnormally elevated,which was related to t

关 键 词：免疫性血小板减少症 NLRP3炎症小体 M1型巨噬细胞 

分 类 号：R725.5[医药卫生—儿科]