肿瘤坏死因子相关凋亡诱导配体对实验性结肠炎小鼠Th17/Treg细胞平衡的影响  

Influence of Tumor Necrosis Factor-related Apoptosis-inducing Ligand on Balance of Th17 and Treg Cells in Mice With Experimental Colitis

作　　者：夏盛隆[1] 金颖莉 吴利敏 卢光荣[1] 钟金伟 全多多 郑波[1] XIA Shenglong;JIN Yingli;WU Limin;LU Guangrong;ZHONG Jinwei;QUAN Duoduo;ZHENG Bo(Department of Gastroenterology,the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou,Zhejiang Province,325000;Department of Pathology,the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou,Zhejiang Province,325000)

机构地区：[1]温州医科大学附属第二医院,育英儿童医院消化内科,325000 [2]温州医科大学附属第二医院,育英儿童医院病理科,325000

出　　处：《胃肠病学》2020年第3期145-150,共6页Chinese Journal of Gastroenterology

基　　金：温州市科技局资助项目(Y20150161)。

摘　　要：背景:Th17/Treg细胞失衡可能是参与克罗恩病(CD)发生的重要环节。肿瘤坏死因子相关凋亡诱导配体(TRAIL)信号通路是影响包括CD在内的多种自身免疫性疾病的潜在机制之一。目的:探讨TRAIL基因敲除对实验性结肠炎小鼠Th17/Treg细胞平衡的影响。方法:采用CRISPR/Cas9基因敲除技术获得TRAIL-/-C57BL/6小鼠。分别将TRAIL-/-小鼠和同品系野生型(WT)小鼠各20只随机分为两组,一组不予处理,一组以三硝基苯磺酸(TNBS)诱导实验性结肠炎。评估各组小鼠结肠炎症程度,采用real-time PCR、蛋白质印迹法和ELISA法检测外周血和结肠组织中的Th17、Treg细胞相关转录因子RORγt、Foxp3和细胞因子白细胞介素-17(IL-17)、IL-10表达。结果:与未造模小鼠相比,造模小鼠体质量显著下降,结肠缩短,疾病活动指数评分和组织活动度指数评分升高(P<0.05),TRAIL-/-结肠炎小鼠上述表现更为明显(P<0.05)。WT结肠炎小鼠外周血和结肠组织中的RORγt、IL-17表达较未造模WT小鼠显著升高(P<0.05),Foxp3、IL-10表达显著降低(P<0.05)。TRAIL-/-结肠炎小鼠上述Th17、Treg细胞相关因子表达和RORγt/Foxp3比值均较WT结肠炎小鼠显著升高(P<0.05)。结论:TRAIL基因敲除可能通过Th17/Treg细胞比例上调加重实验性结肠炎小鼠的肠道炎症反应。Background:Imbalance of Th17/Treg cells might play a key role in the initiation of Crohn’s disease(CD).Moreover,tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)pathway was suggested to be implicated in the pathogenesis of CD and other autoimmune diseases.Aims:To investigate the influence of TRAIL gene deletion on the balance of Th17 and Treg cells in mice with experimental colitis.Methods:TRAIL-/-C57BL/6 mice were obtained by CRISPR/Cas9 method.Then 20 TRAIL-/-mice and 20 wild-type(WT)mice were randomly divided into WT group,TRAIL-/-group,WT trinitrobenzenesulfonic acid(TNBS)-induced colitis group and TRAIL-/-TNBS-induced colitis group,with 10 mice in each group.The severity of colonic inflammation was assessed by disease activity index(DAI)and histology activity index(HAI).Peripheral blood and colonic tissue were collected to determine the expression levels of Th17 and Treg cell-associated transcription factors(RORγt,Foxp3)and cytokines[interleukin-17(IL-17),IL-10]by real-time PCR,Western blotting and ELISA method,respectively.Results:Compared with untreated mice,mice with experimental colitis showed decreased body weight,shortened colon length as well as increased DAI and HAI(P<0.05),and the manifestations of colitis in TRAIL-/-mice were more serious than that in WT mice(P<0.05).In WT mice with experimental colitis,the expression levels of RORγt and IL-17 in peripheral blood and colonic tissue were significantly increased than those in untreated WT mice(P<0.05),while the expression levels of Foxp3 and IL-10 were significantly decreased(P<0.05).Furthermore,the expression levels of all four Th17 and Treg cell-associated molecules were higher in TRAIL-/-colitis mice than in WT colitis mice(P<0.05).In addition,the ratio of RORγt to Foxp3 were higher in TRAIL-/-colitis mice than in WT colitis mice(P<0.05).Conclusions:Deletion of TRAIL gene may aggravate the severity of colonic inflammation via up-regulating Th17/Treg ratio in mice with experimental colitis.

关 键 词：TNF相关凋亡诱导配体 TH17细胞 T淋巴细胞 调节性 结肠炎 

分 类 号：R73[医药卫生—肿瘤]