死亡受体3基因缺陷对结肠炎小鼠肠黏膜炎症及通透性的影响  

Influence of death receptor 3 gene deficiency on the intestinal mucosal inflammation and permeability in colitis mice

作　　者：叶月芳 周刚[2] 庄振杰[3] 付金龙[1,2] 祝思慧[1,2] 朱羽琪[1,2] 李国栋 贺美家 姚金妙 Ye Yuefang;Zhou Gang;Zhuang Zhenjie;Fu Jinlong;Zhu Sihui;Zhu Yuqi;Li Guodong;He Meijia;Yao Jinmiao(School of Clinical Medicine,Hangzhou Normal University,Hangzhou 310015,China;Department of Gastroenterology,Affiliated Hospital of Hangzhou Normal University,Hangzhou 310015,China;Translational Medicine Center,Affiliated Hospital of Hangzhou Normal University,Hangzhou 310015,China)

机构地区：[1]杭州师范大学临床医学院,310015 [2]杭州师范大学附属医院消化内科,310015 [3]杭州师范大学附属医院转化医学平台,310015

出　　处：《中华炎性肠病杂志（中英文）》2021年第4期334-341,共8页Chinese Journal of Inflammatory Bowel Diseases

基　　金：浙江省自然科学基金项目(LY16H030017);杭州市科技发展计划项目(20120633B27);杭州师范大学本科生创新项目(CX2020167、CX2020187)。

摘　　要：目的:分析死亡受体3(Dr3)基因缺陷对不同结肠炎小鼠模型肠黏膜炎症的影响,并探讨Dr3基因缺陷与肠黏膜通透性的关系。方法:收集雌性Dr3基因缺陷(Dr3^(-/-))小鼠和野生型(WT)小鼠各9只,分别设为Dr3^(-/-)-DSS组和WT-DSS组。两组小鼠均采用2.5%葡聚糖硫酸钠(DSS)溶液连续饮用5 d、灭菌水饮用2 d为1个循环,共4个循环的方法建立慢性结肠炎模型。收集雄性WT、Dr3^(-/-)小鼠作为供体鼠,采用磁珠和流式细胞术分别筛选出WT、Dr3^(-/-)的初始T淋巴细胞并腹腔注射至免疫缺陷(Rag1-/-)小鼠和Dr3^(-/-)Rag1-/-小鼠,作为WT转移组和Dr3^(-/-)转移组,从而建立T细胞过继转移诱导的结肠炎模型。观察小鼠体质量、大便性状及潜血,并计算疾病活动指数(DAI)。显微镜下观察小鼠肠黏膜损伤及炎症细胞浸润程度,并计算肠炎组织学评分。通过FITC-dextran血清荧光法检测小鼠肠黏膜通透性。比较两组间的DAI评分、组织学评分和FITC-dextran含量的差异。结果:(1)Dr3^(-/-)-DSS组小鼠的DAI评分在建模第12、19、26天明显高于WT-DSS组小鼠,差异有显著统计学意义(均P<0.05)。建模后4周,WT-DSS组小鼠直肠组织学评分明显高于盲肠和结肠(10.130±1.540比3.667±0.236和7.222±1.199,均P<0.05),提示WT-DSS组直肠炎症程度最重;Dr3^(-/-)-DSS组小鼠结肠组织学评分明显高于盲肠和直肠(11.330±1.167比7.556±1.519和9.500±0.824,均P<0.05),提示Dr3^(-/-)-DSS组结肠炎症程度最重;与WT-DSS组比较,Dr3^(-/-)-DSS组小鼠盲肠和结肠的组织学评分显著增高(盲肠:7.556±1.519比3.667±0.236,P=0.022;结肠:11.330±1.167比7.222±1.199,P=0.026),而两组直肠的组织学评分差异无统计学意义(P>0.05),提示Dr3基因缺陷使盲肠和结肠炎症加重。(2)建模后6周,WT转移组直肠组织学评分明显高于十二指肠、空肠、回肠末端、盲肠、结肠中段(均P<0.05),提示WT转移组直肠炎症最重;Dr3^(-/-)转移组盲肠组织学评�Objective To investigate the influence of death receptor 3(Dr3)gene deficiency on the intestinal mucosal inflammation in different mice colitis models,and explore the relationship of Dr3 gene deficiency and intestinal mucosal permeability.Methods Nine female Dr3 gene deficiency(Dr3^(-/-))mice and 9 wild type(WT)mice were collected and set as Dr3^(-/-)-DSS group and WT-DSS group.The mice of 2 groups received 2.5%dextran sodium sulfate(DSS)for 5 days and sterile water for 2 days as a cycle and 4 cycles were manipulated to construct a chronic colitis model of mice.The male WT and Dr3^(-/-)mice were collected as donor mice and initial T lymphocytes from two types of donor mice were sorted respectively by immunomagnetic separation and flow cytometry.A enteritis model of mice induced by T cells adoptive transfer was constructed on the recipient mice including Rag1-/-(WT transfer group)and Dr3^(-/-)Rag1-/-(Dr3^(-/-)transfer group)mice by the peritoneal injection of T lymphocytes from WT and Dr3^(-/-)mice respectively.The body mass,stool property and occult blood of mice were observed,and the disease activity index(DAI)was calculated.The degree of intestinal mucosal injury and inflammatory cell infiltration in mice were observed under microscope,and the histological score of enteritis was calculated.The intestinal mucosal permeability of mice was detected by fluorescein isothiocyanate(FITC)-dextran serum fluorescence method.The differences of DAI score,histological score and FITC-dextran content between the two groups were compared.Results The DAI scores of mice in Dr3^(-/-)-DSS group were significantly higher than those in WT-DSS group on the 12th,19th and 26th day after establishing the model(all P<0.05).The rectal histological score of WT-DSS group 4 weeks after establishing the model was significantly higher than that of cecum and colon(10.130±1.540 vs.3.667±0.236 and 7.222±1.199,all P<0.05),suggesting that the degree of rectal inflammation in WT-DSS group was the most serious.The histological score of colon in Dr

关 键 词：基因 死亡受体3 结肠炎 肠黏膜通透性 小鼠 炎症性肠病 

分 类 号：R574.62[医药卫生—消化系统]