ConA重复给药建立ACLF免疫状态动态转变的小鼠模型  

Establishment of Model of Dynamic Change of Immune Status of ACLF Induced by ConA Repeated Administration in Mice

作　　者：张楠楠[1] 杨淑殷[1] 陈柳莹[1] 尹珊[1] 王十锦 刘三海 王蓓蓓[2,3] 汪铮[1] 李海[1] 

机构地区：[1]上海交通大学医学院附属仁济医院消化科上海市消化疾病研究所,200001 [2]首都医科大学附属北京地坛医院传染病研究所 [3]首都医科大学新发突发传染病研究北京市重点实验室

出　　处：《胃肠病学》2016年第6期326-330,共5页Chinese Journal of Gastroenterology

基　　金：国家自然基金项目(30971333、81170421、81470869)

摘　　要：背景:慢加急性肝衰竭(ACLF)是我国常见的肝衰竭类型,目前尚缺乏能有效模拟ACLF免疫状态动态转变的动物模型。目的:通过刀豆球蛋白A(ConA)重复给药,建立模拟ACLF免疫状态动态转变的动物模型。方法:小鼠随机分为对照组和ConA重复给药组,ConA重复给药组小鼠给予球后内眦静脉丛注射ConA 15 mg/kg,每隔48 h一次,共5次,对照组给予等体积0.9%NaCl溶液。CBA法检测外周血IL-6、IL-10、IL-12、TNF-α、IFN-γ、MCP-1水平,并测定IL-10/TNF-α比值;流式细胞术检测外周血中单核细胞HLA-DR表达、CD4^+T细胞数量及其比例以及PD-1表达。结果:随着给药次数增加,ConA重复给药组小鼠外周血细胞因子从促炎细胞因子为主转变成抗炎细胞因子为主。与对照组相比,ConA重复给药组外周血中单核细胞HLA-DR表达下降(P<0.05);CD4^+T细胞数量和比例下降(P<0.05),PD-1表达上调(P<0.05)。结论:本研究通过ConA重复刺激成功建立了模拟ACLF免疫状态从全身炎症反应综合征(SIRS)到代偿性抗炎反应综合征(CARS)动态转变的动物模型。Background： Acute-on-chronic liver failure （ACLF） is a commonly seen liver failure in China, and lacking an animal model that can effectively simulate the dynamic change of immune status of ACLF. Aims： To establish an animal model that can simulate dynamic change of immune status of ACLF by repeated administration of coneanavalin A （ConA）. Methods： Mice were randomly divided into normal control group and ConA repeated administration group. Mice in ConA repeated administration group were injected with ConA 15 mg/kg through retrobulbar angular vein every 48 hours for 5 times, and mice in control group were injected with same volume of 0.9% NaC1 solution. Serum levels of IL-6, IL-10, IL- 12, TNF-a, IFN-y/, MCP-1 in peripheral blood were assessed by CBA assay, and the ratio of IL-10/TNF-a was calculated. The expression of HLA-DR, number and proportion of CD4 ＋ T cells and the expression of PD-1 of monocytes in peripheral blood were detected by flow cytometry. Results： Peripheral blood cytokines changed from predominated proinflammatory cytokines into predominated anti-inflammatory cytokines with the increasing in time of administration in ConA repeated administration group. Compared with control group, HLA-DR expression of monocytes in peripheral blood was significantly decreased （ P 〈0. 05 ）, number and proportion of CD4＋ T cells were significantly decreased （ P 〈 0.05 ）, and PD-1 expression was significantly increased （ P 〈 0.05 ） in ConA repeated administration group. Conclusions ： An animal model of ACLF immune status from systemic inflammatory response syndrome （SIRS） to compensatory antiinflammatory response syndrome （CARS） induced by repeated ConA stimulation is successfully established.

关 键 词：慢加急性肝衰竭 伴刀豆球蛋白A 免疫状态 细胞因子类 模型 动物 

分 类 号：R575.3[医药卫生—消化系统] R-332[医药卫生—内科学]