白细胞介素-33对脓毒症小鼠脾脏树突状细胞免疫功能的影响  

Effects of interleukin-33 on the immune dysfunction of dendritic cells in septic mice

作　　者：阴月 王丽雪 董宁[2] 吴瑶 童亚林 祝筱梅[2] 姚咏明[2] Yin Yue;Wang Lixue;Dong Ning;Wu Yao;Tong Yalin;Zhu Xiaomei;Yao Yongming(College of Life Science,Guangxi Normal University,Guilin Guangxi,541006,China;Wound Repair and Regeneration Research Center,Medical Innovation Research Center of the Chinese PLA General Hospital,Beijing 100048,China;Department of Burn and Plastic Surgery,924th Hospital of PLA,Guilin Guangxi,541002,China)

机构地区：[1]广西师范大学生命科学学院,广西桂林541006 [2]解放军总医院医学创新研究部创伤修复与组织再生研究中心,北京100048 [3]解放军联勤保障部队第九二四医院烧伤整形科,广西桂林541002

出　　处：《感染．炎症．修复》2019年第4期247-252,共6页Infection Inflammation Repair

基　　金：国家自然科学基金资助项目(81730057,81871557);国家重点研发计划项目(2017YFC1103302);军队“十三五”医学创新工程重点项目(18CXZ026);漓江学者计划项目。

摘　　要：目的:探讨白细胞介素(IL)-33对小鼠脾脏树突状细胞(DC)功能的影响及其与脓毒症免疫抑制的关系。方法:小鼠随机分为3组:盲肠结扎穿孔(CLP)组采用CLP方法复制脓毒症小鼠模型;假伤组除不行盲肠穿孔外,其他处理方法同CLP组;可溶性IL-33受体(sST2)干预组于CLP术后1 h经腹腔注射重组sST2(每只10μg,溶于200μl PBS中),同批对照CLP组同时给予200μl PBS腹腔注射。应用免疫磁珠分离提纯小鼠脾脏DC。正常小鼠脾脏DC体外培养并给予不同浓度(0、1、10、100 ng/ml)、不同时间(12和24 h)重组IL-33刺激。应用流式细胞术检测各样本DC表面标志、共刺激分子CD80、CD86、主要组织相容性复合物-Ⅱ(MHC-Ⅱ)表达水平。Westernblot检测小鼠脾脏IL-33蛋白表达。结果:与假伤组比较,CLP小鼠脾脏组织IL-33蛋白水平术后24~72 h明显升高,虽然DC表面CD80表达水平有所升高,但CD86、MHC-Ⅱ表达水平均显著降低(P<0.01)。CLP术后1h给予sST2以中和IL-33可改善DC功能状态,DC表面CD80、CD86、MHC-Ⅱ表达水平均显著增强(P<0.01),小鼠存活率也有一定程度的提高。正常小鼠脾脏DC体外培养,重组IL-33体外刺激(10 ng/ml,24 h)可直接诱导DC表面CD86、MHC-Ⅱ表达降低(P<0.05),与CLP模型小鼠脾脏DC体内改变趋势相似。结论:IL-33可直接诱导DC成熟分化障碍,是引发脓毒症小鼠免疫细胞功能抑制的重要因素;拮抗IL-33生物效应可有效地改善DC功能状态,对脓毒症动物发挥保护作用。Objective:To investigate the potential effect of interleukin(IL)-33 on the immune response of mouse spleen dendritic cells(DC)and its relationship with immunosuppression in sepsis.Methods:Mice were randomly divided into three groups.The cecal ligation and perforation(CLP)mouse model was used to mimic sepsis(CLP group).Mice in sham group were treated with the same methods as CLP group,except for cecal perforation.In the sST2 group,recombinant sST2(soluble receptor of IL-33)was injected intraperitoneally within 1 hour after CLP procedure(10μg/mouse,in 200μl PBS),and the same volume of PBS as that of sST2 group was injected in the CLP group.DCs were isolated and purified from mice spleens with immunomagnetic beads.The splenic DCs of normal mice were cultured in vitro and stimulated with recombinant IL-33 at different concentrations(0,1,10,100 ng/ml)and at different times(12 and 24 hours).The expressions of CD80,CD86,and major histocompatibility complex(MHC)-Ⅱon the surface of DCs in every group were determined with flow cytometry.The protein level of IL-33 in mice spleens was detected by Western blotting.Results:Compared with the sham group,levels of IL-33 protein in spleen tissue of CLP mice increased significantly 24-72 hours after procedure.Although the expression level of CD80 on surface of DC increased,levels of CD86 and MHC-Ⅱwere markedly down-regulated(P<0.01).Treatment with sST2 could promote the expression of related markers on the surface of DC that decreased in CLP mice(P<0.01),and the survival rate of CLP mice also increased.In addition,stimulation of recombinant IL-33 induced the down-regulation of CD86 and MHC-Ⅱon surface of splenic DC from normal mice and cultured in vitro(P<0.05).Conclusions:IL-33 has a direct impact on the disturbance of DC maturation and differentiation,which might be critically involved in mediated immune dysfunction in the setting of sepsis.Antagonizing the biological effect of IL-33 can effectively improve the function of DC and play a protective role in sepsis animals.

关 键 词：脓毒症 免疫障碍 白细胞介素-33 树突状细胞 

分 类 号：R392[医药卫生—免疫学]