IL-12单克隆抗体通过调节肠黏膜免疫缓解克罗恩病模型小鼠肠道炎症  

IL-12 monoclonal antibody alleviates intestinal inflammation in IL-10^(-/-) mice by regulating intestinal mucosal immunity

作　　者：王娟 张朝阳 邵志林 王畏 周昌旻 周杰 何逸凡 沈梦迪 葛思堂 左芦根[6] 钟春生 WANG Juan;ZHANG Chaoyang;SHAO Zhilin;WANG Wei;ZHOU Changmin;ZHOU Jie;HE Yifan;SHEN Mengdi;GE Sitang;ZUO Lugen;ZHONG Chunsheng(Medical oncology of the Bengbu Third People's Hospital of Bengbu Medical College,Bengbu 233000;Medical Surgical Oncology of the First Affiliated Hospital of Bengbu Medical College,Bengbu 233004;Emergency Department of the First Affiliated Hospital of Bengbu Medical College,Bengbu 233004;General Surgery of the Second Affiliated Hospital of Bengbu Medical College,Bengbu 233000;Anhui Key Laboratory of Tissue Transplantation of Bengbu Medical College Research Center,Bengbu 233030;General Surgery of the First Affiliated Hospital of Bengbu Medical College,Bengbu 233004,China)

机构地区：[1]蚌埠医学院附属蚌埠市第三人民医院肿瘤内科,安徽蚌埠233000 [2]蚌埠医学院第一附属医院肿瘤外科,安徽蚌埠233004 [3]蚌埠医学院第一附属医院急诊科,安徽蚌埠233004 [4]蚌埠医学院第二附属医院普外科,安徽蚌埠233000 [5]蚌埠医学院科研中心组织移植安徽省重点实验室,安徽蚌埠233030 [6]蚌埠医学院第一附属医院普外科,安徽蚌埠233004

出　　处：《细胞与分子免疫学杂志》2018年第8期678-683,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：蚌埠医学院自然科学基金(BYKY1721ZD)

摘　　要：目的观察白细胞介素12单克隆抗体(IL-12 mAb)对IL-10敲除(IL-10 KO)的克罗恩病(CD)小鼠模型肠道炎症的治疗作用并分析可能的机制。方法选取15周龄雄性IL-10 KO小鼠16只,随机分为对照组及IL-12 mAb处理组。IL-12 mAb处理组小鼠给予腹腔注射IL-12 mAb(25 mg/kg,每周1次),对照组给予0. 2 m L生理盐水腹腔注射。处理4周后,采用炎症性肠病疾病活动度指数(DAI)及HE染色结合Spencer结肠炎组织学评分评估两组小鼠肠道炎症程度及组织学改变;采用活体肠黏膜异硫氰酸荧光素标记的葡聚糖(FITC-dextran)通透性实验评估两组小鼠肠黏膜屏障功能,并采用Western blot法检测肠黏膜密封蛋白1(claudin-1)的蛋白水平;采用流式细胞术检测肠黏膜CD3、CD4、γ干扰素(IFN-γ)和IL-4,评估Th1细胞/Th2细胞平衡。采用ELISA检测两组小鼠肠黏膜IL-13及肿瘤坏死因子α(TNF-α)水平,Western blot法检测肠黏膜磷酸化的信号转导子与转录激活子6(p-STAT6)的蛋白水平。结果 IL-12 mAb治疗后第3周及第4周,IL-12 mAb处理组小鼠DAI及肠道炎症评分均显著低于对照组。同时,IL-12 mAb处理组小鼠肠黏膜通透性显著低于对照组,且肠黏膜claudin-1表达显著高于对照组。同时,IL-12 mAb治疗抑制肠黏膜Th1细胞比例,上调Th2细胞比例。信号通路分析中发现,IL-12 mAb治疗提高肠黏膜p-STAT6及IL-13水平,而抑制TNF-α水平。结论 IL-12 mAb可有效缓解CD小鼠模型的肠道炎症并保护肠黏膜屏障,可能与抑制肠黏膜Th1细胞免疫反应及上调STAT6信号有关。Objective To evaluate the therapeutic effect and possible mechanism of IL-12 monoclonal antibody (IL-12 mAb)on IL-10 knockout(IL-10-/-)mice and its possible mechanism.Methods Sixteen male IL-10^-/-mice of 15weeks old were randomly divided into control group and IL-12 mAb treatment group.The IL-12 mAb treatment group were given intraperitoneal injection of IL-12 mAb (25mg/kg,once per week),and the control group was given intraperitoneal injection of 0.2mL of normal saline.After 4 weeks of intervention,the inflammatory bowel disease activity index (DAI)and HE staining were used to evaluate the intestinal inflammation symptoms and histological changes.The intestinal mucosal permeability test was used to evaluate the intestinal mucosal barrier function of the two groups.The expression of claudin-1 in intestinal mucosawas detected by Western blot analysis.The Th1/Th2 cell balance of intestinal mucosa was evaluated by flow cytometry.The ELISA was used to evaluate IL-13 and tumor necrosis factor alpha(TNF-α)of intestinal mucosal of the two groups.The expression of phosphorylated signal transducer and activator of transcription (p-STAT6)in intestinal mucosa was detected by Western blot nanlysis.Results Three and 4weeks after IL-12 mAb treatment,the DAI and intestinal inflammation scores of IL-12 mAb treatment group were significantly lower than the control group.At the same time,the intestinal mucosal permeability of IL-12 mAb treatment group was significantly lower than that of the control group,and the expression of claudin-1in intestinal mucosa was significantly higher than that of the control group.At the same time,IL-12 mAb treatment inhibited the proportion of Thl cells in the intestinal mucosa and up-regulated the proportion of Th2 cells.In the signal pathway analysis,IL-12 mAb treatment increased the levels of p-STAT6 and IL-13 in the intestinal mucosa and inhibited the level of TNF-α.Conclusion IL-12 mAb effectively alleviates intestinal inflammation in the Crohn's disease animal model and protect the intestin

关 键 词：克罗恩病 白细胞介素12(IL-12) 肠黏膜屏障 炎症 T细胞免疫反应 

分 类 号：R364.5[医药卫生—病理学] R516.1[医药卫生—基础医学]