基础研究miR-19a对溃疡性结肠炎的作用机制.pdfVIP

J South Med Univ, 2013, 33(9): 1325-1328 doi 10.3969/j.issn. 1673-4254.2013.09.15 ·1325 · 基础研究 miR-miR-1919aa 对溃对溃疡性结肠炎的作用机制疡性结肠炎的作用机制 1，3 2 1 3 周鹏志 ，陈 斌 ，胡品津 ，孙 嫣 1 2 中山大学附属第一医院消化科，广东 广州 510089 ；广州中医药大学第一附属医院消化科，广东 广州 3 510405 ； 广州医学院第三附属医院消化科，广东 广州 510150 摘要：目的探讨miR- 19a 对溃疡性结肠炎的作用机制。方法通过生物信息学分析预测miR- 19a 可能的靶基因，通过免疫组化 及Western blotting 技术检测靶基因在溃疡性结肠炎小鼠中表达的变化，并进一步通过绿色荧光蛋白报告载体实验对靶基因进 行鉴定。结果生物信息学分析预测miR- 19a 可能的靶基因是TNF-α，免疫组化及Western blotting 显示溃疡性结肠炎小鼠肠道 组织TNF-α表达增加，miR- 19a 可抑制TNF-α-3UTR-WT 报告基因活性，而变异型TNF-α-3UTR-mut 报告基因活性不能被抑 制。结论miR- 19a 的靶基因为TNF-α，其结合位点为TNF-α3UTR ，miR- 19a 可能在肠道通过直接调控TNF-α而发挥作用。 关键词：溃疡性结肠炎；miR- 19a ；肿瘤坏死因子-α Role of miR-19a in ulcerative colitis in mice 1, 3 2 1 3 ZHOU Pengzhi , CHEN Bin , HU Pinjin , SUN Yan 1Department of Gastroenterology, First Affiliated Hospital, Sun Yat-sen University, Guangzhlu 510089, China; 2Department of Gastroenterology, First Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou 510405, China; 3Department of Gastroenterology, Third Affiliated Hospital, Guangzhou Medical College, Guangzhou 510150, China Abstract: Objective To study the role of miR-19a in ulcerative colitis (UC) in mice. Methods The target gene of miR-19a was predicted by bioinformatics analysis. The expression of the target protein in UC colon was detected by immunohistochemistry and Western blotting. The target gene was further identified by enhanced green fluorescent protein (EGFP) report vector system. Results The target gene of miR-19a was TNF-αas predicted by bioinformatics analysis. TNF-αexpression was highly expressed in the colonic tissue of UC mice. MiR-19a could inhibit the report gene activity of TNF-α-3UTR-WT but no that of TNF-α-3UTR-Mut. Conclusion The targ