Tim-3/aPKC-ι通路在英夫利西单抗治疗克罗恩病病程判断中的作用  

Role of Tim-3/aPKC-ι pathway in evaluating the course of Crohn’s disease treated by infliximab

作　　者：孙华文[1] 王秋爽[1] 周佳伟 SUN Huawen;WANG Qiushuang;ZHOU Jiawei(Dept.of Gastrointestinal Surgery,Renmin Hospital of Wuhan University,Wuhan 430060,Hubei,China)

机构地区：[1]武汉大学人民医院胃肠外科,湖北武汉430060

出　　处：《武汉大学学报（医学版）》2022年第4期658-663,共6页Medical Journal of Wuhan University

基　　金：国家自然科学基金资助项目(编号:81170368)。

摘　　要：目的:探讨克罗恩病(CD)患者接受英夫利西(IFX)治疗前后Tim-3/aPKC-ι通路的变化并分析其临床意义和在病程判断中的作用。方法:制备外周血T细胞亚群同时构建靶向RNA干扰Tim-3基因的慢病毒载体,体外转染T细胞亚群,检测细胞因子分泌水平和Tim-3以及aPKC-ιmRNA的变化,确认Tim-3/aPKC-ι通路的存在;收集2009年6月至2019年7月武汉大学人民医院符合研究标准的45例CD患者,分别在第0、2、6周时给予IFX治疗。以同期16例健康体检者为健康对照组。CD患者于治疗开始前(0周)和治疗第5、10和15周时取静脉血,并经结肠镜检查取肠黏膜活组织检查标本。观察CD患者克罗恩病活动指数(CDAI)、血沉(ESR)、C-反应蛋白(CRP)的变化。使用荧光定量PCR方法分析外周血和肠黏膜组织内Tim-3和aPKC-ιmRNA基因表达水平变化。结果:RNA干扰Tim-3基因在T细胞表达后,可抑制下游的aPKC-ι mRNA表达水平以及T细胞增殖的作用,提示CD发病机制中存在Tim-3/aPKC-ι通路。IFX治疗前,CD患者在外周血Tim-3和aPKC-ι mRNA表达水平(5.25±0.89和5.11±0.45)和肠黏膜组织内Tim-3和aPKC-ι mRNA表达水平(5.77±0.56和5.07±0.35)均明显高于健康对照者,差异均有统计学意义(均P<0.05)。第10周和15周时,CD患者外周血Tim-3和aPKC-ιmRNA表达水平和肠黏膜组织内Tim-3和aPKC-ι mRNA表达水平均明显低于治疗前,差异均有统计学意义(P<0.05)。结论:证实Tim-3/aPKC-ι通路在CD的发病机制中有重要作用,活动期CD患者经IFX治疗后Tim-3和aPKC-ιmRNA表达水平下降,提示Tim-3/aPKC-ι通路参与IFX治疗活动期CD,可以导致临床缓解,下降的幅度对病程判断具有临床意义。Objective:To explore the changes of the Tim-3/aPKC-ι pathway before and after treatment with Infliximab(IFX)in patients with Crohn's disease(CD),and analyze its clinical significance and its role during the course of the disease.Methods:We had prepared peripheral blood T cell subsets and constructed a lentiviral vector that targeting RNA interference Tim-3 gene,transfected T lymphocyte subsets in vitro.The cytokine secretion levels and changes on Tim-3 and aPKC-ι mRNA were detected,and the existence of Tim-3/aPKC-ι pathway was confirmed.A total of 45 CD patients who met the research criteria in Renmin Hospital of Wuhan University from June 2009 to July 2019 had been collected and conducted treatment with IFX at the 0th,2nd,and 6th weeks,respectively.The 16healthy people in the same period served as the healthy control group.In patients with CD,venous blood was collected before the start of treatment(week 0)and at the 5th,10th,and 15th weeks of treatment,and colonoscopy was performed to take biopsy specimens of the intestinal mucosa.The changes of Crohn's disease activity index(CDAI),erythrocyte sedimentation rate(ESR),and C-reactive protein(CRP)in CD patients were observed.The fluorescence quantitative PCR method was used to analyze the changes in the expression levels of Tim-3 and aPKC-ι mRNA genes in peripheral blood and intestinal mucosa.Results:RNA interference with Tim-3 gene expression in T cells could inhibit downstream aPKC-ι mRNA gene expression and T cell proliferation,suggesting that there is a Tim-3/aPKC-ι pathway in the pathogenesis of CD.Before IFX treatment,CD patients had higher expression of Tim-3 and aPKC-ιgenes in peripheral blood(5.25±0.89 and 5.11±0.45)and in the intestinal mucosa(5.77±0.56 and 5.07±0.35),and the levels were significantly higher than those of healthy controls(all P<0.05).After 10 and 15 weeks'treatment,the expression of Tim-3 and aPKC-ι mRNA in peripheral blood and in intestinal mucosa tissue of CD patients were significantly lower than before treatment(all P<0.05

关 键 词：Tim-3/aPKC-ι通路 英夫利西单抗 克罗恩病 

分 类 号：R574.62[医药卫生—消化系统]