南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (12): 2023-2028.doi: 10.12122/j.issn.1673-4254.2023.12.05

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UBE2W过表达促进结肠炎小鼠结肠细胞的增殖

王少鑫，崔立红，李 辉，刘新尧，李晓伟，王晓辉   

  1. 解放军总医院第六医学中心消化内科，北京 100048
  • 出版日期:2023-12-20 发布日期:2023-12-29

UBE2W overexpression promotes proliferation of intestinal mucosal cells in mice with chemically induced colitis

WANG Shaoxin, CUI Lihong, LI Hui, LIU Xinyao, LI Xiaowei, WANG Xiaohui   

  1. Department of Gastroenterology, Sixth Medical Center of Chinese PLA General Hospital, Beijing 100048, China
  • Online:2023-12-20 Published:2023-12-29

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摘要： 目的 利用结肠炎小鼠模型，探讨UBE2W对结肠炎粘膜细胞增殖的作用。方法 利用2.5%葡聚糖硫酸钠盐（DSS）喂养C57BL/6小鼠构建结肠炎小鼠模型，采用聚合酶链反应（PCR）和Western blot检测小鼠结肠组织UBE2W的表达水平。将结肠炎小鼠随机进行尾静脉注射腺病毒（AAV2/9 myc-UBE2W或AAV2/9 Control），分为UBE2W过表达组和对照组，5只/组。观察小鼠的体质量变化，实验结束时取结肠组织行病理HE染色及组织学评分，评价小鼠结肠炎症的严重程度。采用免疫组织化学方法检测两组小鼠结肠粘膜细胞的增殖指标Ki67和BrdU表达水平。在293T和HCT116细胞中，采用CCK-8试剂盒检测过表达UBE2W对细胞增殖的影响。结果 成功建立由DSS诱导的实验性结肠炎小鼠模型，UBE2W在结肠组织中mRNA和蛋白水平与正常小鼠比较明显降低；UBE2W过表达组结肠炎小鼠体质量丢失少于对照组，在第9天和第10天差异最明显（P<0.05），其病理组织学结肠炎症指数低于对照组（P<0.05）；UBE2W过表达组小鼠结肠组织细胞中Ki67和BrdU表达水平高于对照组（P<0.05）。在293T中过表达UBE2W后细胞培养72、96 h时的吸光度A450 nm均高于对照组（P<0.05），HCT116细胞呈现类似结果。结论 UBE2W对小鼠结肠炎有保护作用，过表达UBE2W有利于结肠炎症的恢复，可能是通过促进结肠细胞增殖实现的。

关键词: 泛素结合酶E2W；葡聚糖硫酸钠盐；结肠炎；细胞增殖

Abstract: Objective To explore the effect of UBE2W overexpression on proliferation of intestinal mucosal cells in a mouse model of dextran sulfate sodium (DSS)-induced colitis. Methods In a mouse model of colitis induced by exposure to DSS in drinking water, UBE2W expression in the colon tissue was detected by RT-PCR and Western blotting. Ten mouse models of colitis were randomized for injection of adenovirus AAV2/9 myc-UBE2W or control AAV2/9 via the tail vein (n=5), and the changes in body weight were recorded and the histological score of the colon were graded using HE staining. Ki67 and BrdU expressions in the colon mucosal cells were detected with immunohistochemistry. The effect of UBE2W overexpression on proliferation of 293T and HCT116 cells was observed using CCK-8 kit. Results Compared with normal mice, the mouse models with DSS-induced colitis showed significantly lowered expressions of UBE2W mRNA and protein in the colon tissues. The mouse models with AAV2/9 myc-UBE2W injection had a lower body weight loss than those with control AAV2/9 injection, and the difference was the most distinct on days 9 and 10 (P<0.05). AAV2/9 myc-UBE2W injection significantly decreased the histological score (P<0.05) and increased Ki67 and BrdU expressions in the colon mucosal cells in the mouse models (P<0.05). In both 293T and HCT116 cells, UBE2W overexpression significantly promoted cell proliferation at 72 h and 96 h after plasmid transfection (P<0.05). Conclusion UBE2W overexpression provide protection of the colon mucosal cells and promotes recovery of colitis in mice possibly by promoting proliferation of the colon mucosal cells.

Key words: ubiquitin-conjugating enzyme E2W; dextran sodium sulfate; colitis; cell proliferation