E3泛素连接酶RNF138抑制溃疡性结肠炎的发生及进展

E3 ubiquitinase RNF138 inhibits the development of ulcerative colitis

目的研究E3泛素连接酶RNF138在溃疡性结肠炎(UC)中的作用。方法利用GEO数据库分析活动期、缓解期以及经免疫抑制剂治疗后UC样本中RNF138的表达水平;利用RNF138基因敲除小鼠构建UC模型,对模型小鼠进行病理学观察及组织学分析;取小鼠结肠组织进行转录组测序,初步探究RNF138敲除促进UC进展的可能机制;利用免疫组化和Western blot检测小鼠结肠组织中RNF138和phospho-NF-κB p65(p-p65)的表达水平;通过qPCR实验检测核因子NF-кB通路靶基因的表达水平;利用免疫组化对临床样本中RNF138和p-p65的表达进行检测。结果GEO数据库及临床样本分析显示,与缓解期相比,活动期UC组织中RNF138表达下调(P<0.05);敲除RNF138促进DSS诱导的UC发生及进展(P<0.05);敲除RNF138促进p-p65及其靶基因表达(P<0.05),提示NF-кB信号通路激活。结论RNF138通过阻断NF-кB通路活化抑制UC发生及进展。

Objective To explore the role of RNF138(ring finger protein 138)in the ulcerative colitis(UC).Methods The expression of RNF138 in active,remission and ulcerative colitis upon immunosuppressive treatment was analyzed by GEO database.The UC model was constructed by RNF138-deficient mice.The pathological observation,histological analysis and colon measurement were performed by the model mice.The transcriptome analysis was employed by using mouse colon tissue samples.The expression levels of RNF138 and p-p65 in colon tissue of model mice were detected by immunohistochemistry and Western blots.The expression of target gene of NF-кB pathway was detected by qPCR.Immunohistochemistry was used to evaluate the expression of RNF138 and NF-кB p65 in clinical samples of UC.Results GEO database and clinical samples showed that the expression of RNF138 decreased in the samples of active UC as compared to that in the remission stage(P<0.05).Knocking out RNF138 promoted the occurrence and progression of DSS induced UC(P<0.05).Knocking out RNF138 promoted upregulation of p-p65 and its target gene expression(P<0.05),indicating NF-кB signal pathway activation.Conclusions RNF138 inhibits the development of UC through suppressing the NF-кB genetic pathway.