ASK1在克罗恩病中的表达及其对肠上皮屏障功能和炎症反应的影响  

The expression of ASK1 and its impact on intestinal epithelial barrier function and inf lammatory response in Crohn's disease

作　　者：朱邢超 项镜蓉 王佳宇 张欢 冯洁 石通国 奚沁华[1] ZHU Xingchao;XIANG Jingrong;WANG Jiayu;ZHANG Huan;FENG Jie;SHI Tongguo;XI Qinhua(Department of Gastroenterology,First Affiliated Hospital of Soochow University,Suzhou 215000,China;Jiangsu Institute of Clinical Immunology,First Affiliated Hospital of Soochow University,Suzhou 215000,China)

机构地区：[1]苏州大学附属第一医院消化内科,215000 [2]江苏省临床免疫研究所

出　　处：《免疫学杂志》2025年第1期36-43,共8页Immunological Journal

基　　金：苏州大学附属第一医院临床研究类项目(0302020301031)。

摘　　要：目的本研究旨在探究凋亡信号调节激酶1(ASK1)在克罗恩病(CD)患者肠组织中的表达模式,并深入分析其对肠上皮屏障功能和炎症反应的影响机制。方法收集36例克罗恩病患者手术切除的回肠组织及健康体检者的回肠组织石蜡标本。利用免疫组织化学染色法观察ASK1蛋白的表达,并分析其与克罗恩病活动指数(CDAI)之间的相关性。此外,建立TNBS诱导的小鼠急性结肠炎模型,通过qRT-PCR、Western blot和免疫组织化学染色法分析ASK1的表达,并探讨其与小鼠疾病活动指数(DAI)的关系。通过慢病毒介导的转染技术,在Caco-2细胞中构建ASK1敲低或过表达的稳定细胞系,并在炎性环境下检测这些细胞跨膜电阻(TEER)值、FITC-dextran渗漏浓度和IL-6、IL-1β的mRNA、蛋白浓度的变化。同时,我们通过qRT-PCR和Western blot检测了ASK1表达变化对Krüppel样因子4(KLF4)表达的影响。结果ASK1在CD患者回肠组织中高表达,并与CDAI呈正相关。在TNBS诱导的小鼠模型中,ASK1表达显著上调,与DAI也呈正相关。在炎性环境下,ASK1过表达削弱而敲低则保护Caco-2细胞构建的单层肠上皮屏障,并且ASK1能促进炎症因子的表达。此外,ASK1敲低促进而过表达则抑制KLF4的表达,显示ASK1与KLF4之间存在负相关性。结论CD中ASK1的表达显著升高,并与患者的疾病活动指数呈正相关。ASK1的干预能够调节肠道屏障的完整性和炎症因子的表达,这一作用可能与其调控KLF4表达相关。Objective To examine the expression pattern of apoptosis signal-regulating kinase 1(ASK1)in the intestinal tissues of patients with Crohn's disease(CD),and analyze its mechanistic impact on intestinal epithelial barrier function and inflammatory responses.Methods Ileal tissue samples from Crohn's disease patients and healthy controls were collected.ASK1 protein level was assessed by immunohistochemistry,and its relationship with the Crohn's disease activity index(CDAI)was analyzed.A mouse model of acute colitis was constructed using TNBS,and subjected to qRTPCR,Western blotting and immunohistochemistry for ASK1 expression,and the association between ASK1 expression and the disease activity index was examined.Lentivirus transfection was employed to create stable Caco-2 cell lines with altered ASK1 expression,and the intestinal barrier integrity and inflammation were assessed by measuring transepithelial electrical resistance(TEER),FITC-dextran leakage,and IL-6,IL-1βlevels.Furthermore,the effects of ASK1 expression on Krüppel-like factor 4(KLF4)levels was examined using qRT-PCR and Western blotting.Results ASK1 was highly expressed in the ileum of CD patients and positively correlated with CDAI.In a TNBS-induced mouse model of acute colitis,ASK1 expression was up-regulated and positively correlated with DAI.Inflammation-induced ASK1 overexpression weakened the Caco-2 cell intestinal barrier,whereas ASK1 knockdown strengthened it.Moreover,ASK1 had the capability to enhance the expression of inflammatory factors.Additionally,ASK1 knockdown increased KLF4 expression,while overexpression decreased it,indicating a negative correlation between ASK1 and KLF4.Conclusion ASK1 expression is notably higher in CD and positively correlates with disease activity.ASK1 can influence intestinal barrier integrity and inflammatory factor expression,possibly through its impact on KLF4 expression.

关 键 词：克罗恩病 凋亡信号调节激酶1 肠上皮屏障 Krüppel样因子4 

分 类 号：R574.62[医药卫生—消化系统]