瑞巴派特通過抑制miR-877-5p的表達減輕阿司匹林對人胃黏膜上皮細胞損傷的研究

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南京醫科大學附屬南京醫院(南京市第一醫院) 1.消化科； 2.中心實驗室，江蘇 南京 210006； 3.解放軍南京總醫院急診科

【Abstract】ObjectiveTo investigate whether Rebamipide plays a protective role on Aspirin-induced injury through inhibiting the expression of miR-877-5p in human gastric epithelial cells (GES-1).MethodsCultured GES-1 cells were divided into control group, Aspirin injured group and different concentrations (0.25, 0.5, 1.0 mmol/L) Rebamipide plus Aspirin groups. The expression of miR-877-5p was detected by qRT-PCR. The Aspirin treated cells were transfected with miR-877-5p inhibitors. The combination of Rebamipide and Aspirin treated cells were transfected with miR-8877-5p mimics. Cell proliferation and apoptosis were detected by CCK-8 and flow cytometry. The targeted genes of miR-877-5p were predicted by miRNA target databases, the GO and KEGG pathway were analyzed by DAVID database.ResultsqRT-PCR showed that the expression of miR-877-5p in Aspirin group was the highest than others. The expressions of miR-877-5p in different concentrations (0.25, 0.5, 1.0 mmol/L) of Rebamipide plus Aspirin groups were 4.28±0.25, 2.45±0.28 and 1.47±0.17. The expression of miR-877-5p was gradually decreased with the increase of concentration of Rebamipide. The CCK-8 assay and flow cytometry showed that miR-877-5p mimics transfection blocked proliferations and promoted apoptosis in combination of Rebamipide and Aspirin treated cells, while miR-877-5p inhibitors promoted proliferation and inhibited apoptosis in Aspirin treated cells. The pathways of miR-877-5p targeted genes were almost focused on cAMP signaling pathway, MAPK signaling pathway, PI3K-Akt signaling pathway, and so on.ConclusionRebamipide alleviates Aspirin-induced injury through inhibiting the expression of miR-877-5p in human gastric epithelial cells.

【Keywords】 Rebamipide; Aspirin; Gastric mucosa damage; miR-877-5p

近年來，隨著阿司匹林在心腦血管疾病中的廣泛應用，其相關不良反應尤其是胃黏膜損傷的發生率逐年上升。阿司匹林相關胃黏膜損傷的臨床表現可以無癥狀或有消化不良、出血、穿孔等。瑞巴派特是臨床上常用的胃黏膜保護劑，廣泛應用于胃黏膜損傷和潰瘍的防治中，尤其在阿司匹林相關胃黏膜損傷的防治中效果突出[2-4]。然而，瑞巴派特對胃黏膜保護作用的機制尚未完全明確，近年來研究[5-6]發現，miRNAs參與了細胞發育、增殖、分化和凋亡等重要過程，這為研究瑞巴派特對胃黏膜保護作用機制提供了新的研究方向。

有研究[7]發現，NSAIDs類藥物引起的細胞損傷中miR-877-5p明顯上調，而我們經生物信息學初步分析發現，其可能參與細胞的增殖和凋亡調控，因此推測NSAIDs藥物阿司匹林對胃黏膜的損傷作用可能與miR-877-5p有關，而胃黏膜保護劑瑞巴派特也可能是通過調控該miRNA表達發揮保護性作用。因此，本研究將從阿司匹林和瑞巴派特對miR-877-5p表達的影響和miR-877-5p對人胃黏膜上皮細胞GES-1增殖、凋亡的影響兩方面入手，闡明瑞巴派特防治阿司匹林相關胃黏膜損傷的作用機制，為瑞巴派特的臨床應用提供更多的理論基礎。

1 材料與方法

1.1材料人胃黏膜上皮細胞株GES-1由南京市第一醫院中心實驗室保存。瑞巴派特(上海阿拉丁生物技術有限公司)，阿司匹林(美國Sigma公司)，脂質體……