摘要/Abstract
摘要： 目的·探究miR-23a-3p和miR-27a-3p在溃疡性结肠炎（ulcerative colitis，UC）小鼠血清中的表达及其可能的作用机制。方法·将20只雄性C57BL/6小鼠随机分为对照组和模型组，每组10只。模型组小鼠采用含有5%硫酸葡聚糖钠（dextran sulfate sodium，DSS）的饮用水诱导7 d，诱导期间观察小鼠一般情况、粪便形态以及隐血状况；7 d后，采集小鼠全血和结肠组织，测量结肠长度并称取湿重。qRT-PCR检测血清中miR-23a-3p和miR-27a-3p的表达，Western blotting检测过氧化物酶体增殖物激活受体γ辅助激活因子1α（peroxisome proliferator-activated receptor γ，coactivator 1α，PPARGC1A）、PH域富含亮氨酸重复蛋白磷酸酶2（PH domain leucine-rich repeat protein phosphatase，PHLPP2）、B淋巴细胞瘤2蛋白（B-cell lymphoma-2，BCL-2）、BCL-2相关X蛋白（Bcl-2 associated X protein，BAX）、细胞色素C（cytochrome c，CYT-C）和胱天蛋白酶3剪切体（cleaved cysteine-containing aspartate-specific proteases，cleaved-caspase-3）蛋白表达。结果·DSS诱导后，模型组小鼠出现精神萎靡、体质量减轻、腹泻、肉眼血便等症状，结肠长度缩短、湿重减轻（均P<0.05），小鼠UC模型构建成功。与对照组比较，模型组小鼠血清中miR-23a-3p和miR-27a-3p表达明显下调（均P<0.05），结肠组织中PPARGC1A、PHLPP2、BAX、CYT-C和cleaved-caspase-3蛋白表达显著升高（均P<0.05），而BCL-2表达显著降低（P<0.05）。结论·miR-23a-3p和miR-27a-3p在UC小鼠血清中呈低表达水平，可能通过介导结肠组织中PPARGC1A和PHLPP2表达上调，触发线粒体途径诱导细胞凋亡，从而参与了UC的发生/发展。
关键词: 溃疡性结肠炎, 疾病活动指数, 微小RNA, 过氧化物酶体增殖物激活受体γ辅助激活因子-1α, PH域富含亮氨酸重复蛋白磷酸酶2, 线粒体途径, 细胞凋亡
Abstract:
Objective · To explore the expression levels of miR-23a-3p and miR-27a-3p in the sera of mice with ulcerative colitis (UC) and their potential action mechanisms. Methods · Twenty C57BL/6 mice were randomly divided into the control group and the model group with 10 mice in each group. The model group mice were induced orally by water with 5% dextran sulfate sodium (DSS) for 7 d. During the induction period, the general condition, fecal morphology and occult blood status of the mice were observed. After 7 d, the whole blood and colon tissues of mice were collected, and the colon lengths and wet weights were measured. The expressions of miR-23a-3p and miR-27a-3p in the sera were detected by qRT-PCR. The expressions of peroxisome proliferator-activated receptor γ, coactivator-1α (PPARGC1A), PH domain leucine-rich repeat protein phosphatase 2 (PHLPP2), B-cell lymphoma-2 (BCL-2), BCL-2 associated X protein (BAX), cytochrome c (CYT-C) and cleaved cysteine-containing aspartate-specific protease (cleaved-caspase-3) were detected by Western blotting. Results · After DSS induction, the model group mice showed mental depression, weight loss, diarrhea, and bloody stool, whose colon lengths were shortened and colon wet weights decreased. The UC model was constructed successfully. In the model group, the expressions of miR-23a-3p and miR-27a-3p in the sera decreased significantly (P<0.05), and the expressions of PPARGC1A, PHLPP2, BAX, CYT-C and cleaved-caspase-3 in the colon tissues increased significantly (P<0.05), but BCL-2 decreased (P<0.05). Conclusion · The expressions of miR-23a-3p and miR-27a-3p are low in the sera of UC mice, which may be involved in the occurrence and development of UC by up regulating the expressions of PPARGC1A and PHLPP2 in the colons and triggering mitochondrial pathway to induce apoptosis.
Key words: ulcerative colitis (UC), disease activity index (DAI), miRNA, peroxisome proliferator-activated receptor γ, coactivator-1α (PPARGC1A), PH domain leucine-rich repeat protein phosphatase 2 (PHLPP2), mitochondrial pathway, cell apoptosis
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