基础医学与临床 ›› 2021, Vol. 41 ›› Issue (7): 946-950.

• 研究论文 • 上一篇    下一篇

敲低Ube2w增加葡聚糖硫酸钠诱导的小鼠溃疡性结肠炎的易感性

王少鑫,崔立红,李晓伟,刘新尧,罗哲,闫志辉   

  1. 解放军总医院第六医学中心
  • 收稿日期:2020-06-19 修回日期:2020-10-08 出版日期:2021-07-05 发布日期:2021-06-17
  • 通讯作者: 崔立红 E-mail:luckycui861@163.com
  • 基金资助:
    一新型泛素结合酶UBE2W对NF-?B信号通路的调控和分子机制研究

Knockdown of Ube2w increases the susceptibility to dextran sodium sulfate-induced ulcerative colitis in mice

  • Received:2020-06-19 Revised:2020-10-08 Online:2021-07-05 Published:2021-06-17

摘要: 目的 探讨泛素结合酶(E2)w(UBE2W)对葡聚糖硫酸钠(DSS)诱导的小鼠结肠炎性反应的影响。方法 在小鼠成纤维细胞L929中转染UBE2W siRNA不同干涉序列,Western blot检测UBE2W蛋白表达,选择干涉效果最佳的siRNA进行小鼠体内转染实验。给10只C57BL/6雄性小鼠随机尾静脉注射siRNA(siUBE2W/siControl),并用2.5%DSS喂养,构建两组DSS小鼠结肠炎模型,即UBE2W敲低组和对照组各5只。通过Western blot和RT-PCR检测小鼠结肠组织UBE2W的表达水平,观察两组小鼠的体重,并通过组织学评分比较结肠炎性反应的严重程度。 结果L929细胞转染后UBE2W siRNA2# 抑制UBE2W蛋白表达最为明显,敲低UBE2W组小鼠结肠组织中UBE2W mRNA和蛋白表达水平均低于对照组,且UBE2W敲低组小鼠体重丢失明显多于对照组,敲低组小鼠结肠炎组织学评分为8.60±0.24分高于对照组5.20±0.49分(P<0.05)。 结论 UBE2W参与了炎性反应的调控过程,UBE2W敲低能够增加DSS诱导的小鼠结肠炎易感性。

关键词: 泛素结合酶E2W, 葡聚糖硫酸钠盐, 结肠炎

Abstract: Objective To investigate the effect of UBE2W on dextran sodium sulfate (DSS) - induced ulceractive colitis in mice. Methods Different interference sequences of UBE2W siRNA (1#, 2# and 3#) were transiently transfected into mouse fibroblast L929. The expression of UBE2W protein was detected by Western blot. The siRNA with best inhibitory effect was selected for transfection in vivo. Ten C57BL / 6 male mice were randomly injected with siRNA (siUBE2W / siControl) via caudal vein and fed with 2.5% DSS to construct two groups of DSS colitis models, namely UBE2W knockdown group and control group with 5 mice in each group. Western blot and RT-qPCR were used to detect the expression of UBE2W in colon tissue of mice. The weight of mice in the two groups was observed, and the severity of colitis was compared by histological score. Results UBE2W siRNA 2# inhibited UBE2W protein expression most significantly in L929 cells. UBE2W mRNA and protein expression levels in colon tissue of UBE2W knockdown group were lower than those in control group, and weight loss in UBE2W knockdown group was significantly more than that in control group, and the histological score of colitis in knockdown group was 8.60 ± 0.24, which was higher than that in control group (5.20 ± 0.49, P < 0.05). Conclusion UBE2W is involved in the regulation of inflammatory response, UBE2W knockdown can increase the susceptibility of DSS induced colitis in mice.

Key words: ubiquitin-conjugating enzyme E2W, dextran sodium sulfate, colitis