， ，， ， ，

南京醫(yī)科大學(xué)附屬南京醫(yī)院(南京市第一醫(yī)院) 1.消化科； 2.中心實驗室，江蘇 南京 210006； 3.解放軍南京總醫(yī)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(yīng)用，其相關(guān)不良反應(yīng)尤其是胃黏膜損傷的發(fā)生率逐年上升。阿司匹林相關(guān)胃黏膜損傷的臨床表現(xiàn)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(yīng)用于胃黏膜損傷和潰瘍的防治中，尤其在阿司匹林相關(guān)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(fā)現(xiàn)，miRNAs參與了細(xì)胞發(fā)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(fā)現(xiàn)，NSAIDs類藥物引起的細(xì)胞損傷中miR-877-5p明顯上調(diào)，而我們經(jīng)生物信息學(xué)初步分析發(fā)現(xiàn)，其可能參與細(xì)胞的增殖和凋亡調(diào)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(guān)，而胃黏膜保護劑瑞巴派特也可能是通過調(diào)控該miRNA表達發(fā)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(xì)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(guān)胃黏膜損傷的作用機制，為瑞巴派特的臨床應(yīng)用提供更多的理論基礎(chǔ)。

1 材料與方法

1.1材料人胃黏膜上皮細(xì)胞株GES-1由南京市第一醫(yī)院中心實驗室保存。瑞巴派特(上海阿拉丁生物技術(shù)有限公司)，阿司匹林(美國Sigma公司)，脂質(zhì)體轉(zhuǎn)染試劑 Lipofectamine 2000(美國 Invitrogen)，miR-877-5p mimics和miRNA negative control(NC)、miR-877-5p inhibitor和miRNA inhibitor negative control(INC)、實時熒光定量試劑盒(上海吉瑪)，細(xì)胞總RNA提取試劑Trizol、CCK-8試劑盒、Annexin V-FITC/PI雙染細(xì)胞凋亡檢測試劑盒(南京翼飛雪生物科技有限公司)。

1.2 方法

1.2.1 細(xì)胞培養(yǎng)：GES-1細(xì)胞用含質(zhì)量濃度為100 g/L胎牛血清的DMEM高糖培養(yǎng)基，在37 ℃、體積分?jǐn)?shù)為5%的CO2條件下常規(guī)培養(yǎng)傳代。

1.2.2 實驗分組：研究瑞巴派特對細(xì)胞中miR-877-5p表達的影響，實驗分為5組：不加藥物干預(yù)的空白對照組(NC組)；加入2.21 mmol/L濃度的阿司匹林培養(yǎng)液(即IC10濃度)處理24 h設(shè)為阿司匹林損傷組[8]；加入不同濃度(0.25、0.5、1.0 mmol/L)瑞巴派特預(yù)處理2 h，再加入IC10濃度的阿司匹林培養(yǎng)液處理24 h，設(shè)為瑞巴派特保護組。實驗重復(fù)3次。

研究miR-877-5p對GES-1細(xì)胞增殖和凋亡的影響，實驗分為4組：1、2組均用終濃度為0.5 mmol/L瑞巴派特聯(lián)合阿司匹林處理24 h后分別加入miR-877-5p mimics、NC進行轉(zhuǎn)染；3、4組均用阿司匹林處理24 h后分別加入miR-877-5p inhibitor、INC進行轉(zhuǎn)染。轉(zhuǎn)染后進行細(xì)胞凋亡實驗和細(xì)胞增殖實驗。

1.2.3 細(xì)胞轉(zhuǎn)染：取對數(shù)生長期接種于細(xì)胞培養(yǎng)板中培養(yǎng)24 h，待細(xì)胞達70%～80%融合時進行轉(zhuǎn)染。按照Lipofectamine 2000說明書進行轉(zhuǎn)染，將miR-877-5p mimics、NC、inhibitor和INC分別轉(zhuǎn)染至細(xì)胞中。6 h后更換新鮮培養(yǎng)基，48 h后收集細(xì)胞或進行相關(guān)實驗，通過實時熒光定量聚合酶鏈?zhǔn)椒磻?yīng)(qRT-PCR)驗證轉(zhuǎn)染是否成功。

1.2.4 qRT-PCR：收集細(xì)胞，提取總RNA，測定RNA濃度并按照說明書進行逆轉(zhuǎn)錄，將逆轉(zhuǎn)錄產(chǎn)物cDNA加入反應(yīng)體系中進行qPCR反應(yīng)，檢測各組細(xì)胞間miRNA表達的差異?；蛳鄬Ρ磉_量用2-ΔΔCt表示。實驗重復(fù)3次，取均值。

1.2.5 細(xì)胞增殖檢測：細(xì)胞鋪于96孔板中，分別繼續(xù)培養(yǎng)24 h、48 h和72 h，每孔加入10 μl CCK-8，37 ℃孵育2 h，以450 nm為測定波長，在酶標(biāo)儀上檢測各孔密度值(OD值)。OD值可反映細(xì)胞增殖情況，每組設(shè)4個復(fù)孔，實驗重復(fù)3次，取均值。

1.2.6 細(xì)胞凋亡檢測：各組細(xì)胞轉(zhuǎn)染48 h后，收集細(xì)胞，PBS洗滌2遍，依次加入500 μl結(jié)合緩沖液及Annexin V FITC和PI各5 μl，混勻后室溫避光孵育15 min，上流式細(xì)胞儀檢測。實驗重復(fù)3次，取均值。

1.2.7 miR-877-5p靶蛋白預(yù)測：通過miRNA靶基因在線預(yù)測軟件(Targetscan、miRWalk、miRanda)，取3個數(shù)據(jù)庫預(yù)測的交集靶基因進行GO功能富集分析和KEGG信號通路富集分析。

2 結(jié)果

2.1miR-877-5p在阿司匹林損傷組和瑞巴派特保護組中的表達情況qRT-PCR檢測miR-877-5p表達量，設(shè)空白對照組表達量為1，阿司匹林損傷組為6.43±0.48,0.25 mmol/L瑞巴派特保護組為4.28±0.25,0.5 mmol/L瑞巴派特保護組為2.45±0.28,1.0 mmol/L瑞巴派特保護組為1.47±0.17,阿司匹林損傷組中miR-877-5p表達高于其他保護組(P<0.001)。不同濃度的瑞巴派特保護組中，miR-877-5p表達量依次降低，呈濃度依賴性(F=71.87,P<0.01)。以上結(jié)果說明,瑞巴派特能夠抑制GES-1細(xì)胞中miR-877-5p的表達。

2.2轉(zhuǎn)染miR-877-5pmimics和inhibitor的效果分析qRT-PCR結(jié)果顯示，miR-877-5p mimics組miR-877-5p的表達是NC組的(92.77±8.00)倍，差異有統(tǒng)計學(xué)意義(P<0.001)；miR-877-5p inhibitor組miR-877-5p的表達是INC組的(0.55±0.02)倍，差異有統(tǒng)計學(xué)意義(P<0.01)。以上結(jié)果說明轉(zhuǎn)染成功。

2.3miR-877-5p對人胃黏膜上皮細(xì)胞GES-1增殖的影響細(xì)胞增殖結(jié)果顯示，轉(zhuǎn)染miR-877-5p mimics組的細(xì)胞增殖能力較NC組顯著降低(P<0.01)，第3天抑制增殖效果最明顯[(0.683±0.035)vs(1.112±0.039),P<0.001]。轉(zhuǎn)染miR-877-5p inhibitor組的細(xì)胞增殖能力較INC組明顯增強(P<0.05)(見表1)。以上結(jié)果提示，miR-877-5p對人胃黏膜上皮細(xì)胞的增殖具有抑制作用。

表1 CCK-8法檢測miR-877-5p對人胃黏膜上皮細(xì)胞GES-1增殖的影響Tab 1 Analysis of proliferation in miR-877-5p transfected GES-1 cells by CCK-8

注：與NC組相比，*P<0.05，**P<0.001；與INC組相比，#P<0.05。

2.4miR-877-5p對人胃黏膜上皮細(xì)胞GES-1凋亡的影響流式細(xì)胞凋亡實驗結(jié)果顯示，NC組、miR-877-5p mimics組、INC組和miR-877-5p inhibitor組細(xì)胞凋亡率分別為(8.93±0.74)%、(24.87±3.35)%、(13.4±0.96)%、(5.87±0.53)%。與NC組比較，miR-877-5p mimics處理的細(xì)胞凋亡率明顯增加，差異有統(tǒng)計學(xué)意義(P<0.01)。與INC組比較，miR-877-5p inhibitor處理的細(xì)胞凋亡率降低，差異有統(tǒng)計學(xué)意義(P<0.05)(見圖1)。以上結(jié)果表明，上調(diào)細(xì)胞內(nèi)miR-877-5p的表達可促進人胃黏膜上皮細(xì)胞的凋亡，而下調(diào)其表達，可抑制細(xì)胞的凋亡。

圖1 流式細(xì)胞術(shù)檢測miR-877-5p對人胃黏膜上皮細(xì)胞凋亡的影響

A：NC組；B：miR-877-5p mimic組；C：INC組；D：miR-877-5p inhibitor組

Fig1AnalysisofapoptosisinmiR-877-5ptransfectedGES-1cellsbyflowcytometry

A: NC group; B: miR-877-5p mimic group; C: INC group; D: miR-877-5p inhibitor group

2.5miR-877-5p靶基因預(yù)測和功能分析應(yīng)用Targetscan、miRWalk和miRanda 3個數(shù)據(jù)庫進行靶基因預(yù)測并取交集基因，共發(fā)現(xiàn)交集基因有945個。對這些靶基因富集進行GO功能富集分析發(fā)現(xiàn)，miR-877-5p靶基因主要參與了金屬離子結(jié)合、轉(zhuǎn)錄調(diào)節(jié)、細(xì)胞內(nèi)蛋白修飾、蛋白定位等生物功能(P<0.05)。對這些靶基因進行KEGG通路分析發(fā)現(xiàn)，共有65個明顯富集的通路(P<0.05)，其中富集指數(shù)較高的通路有cAMP信號通路、鈣離子信號通路、PI3K-Akt信號通路、MAPK信號通路等(見圖2)。

3 討論

miRNA是近年來生命科學(xué)領(lǐng)域研究的一個熱點，它是一類廣泛存在于真核生物體內(nèi)，長度在19～25個核苷酸的內(nèi)源性非編碼RNA。它能通過干擾mRNA翻譯過程即堿基互補配對方式來調(diào)節(jié)靶基因的表達[9]，對體內(nèi)細(xì)胞的增殖、分化、損傷、凋亡等方面發(fā)揮重要作用。隨著近幾年研究的不斷深入，細(xì)胞損傷及凋亡與miRNA的聯(lián)系越來越清晰。多項研究[10-14]發(fā)現(xiàn)，阿司匹林可通過調(diào)節(jié)多種特異性miRNA的表達，在多種疾病如先兆子癇、胃腸道腫瘤等中發(fā)揮作用。既往研究發(fā)現(xiàn)，miR-877-5p參與多種細(xì)胞凋亡的過程[15-18]，在多種藥物引起的肝損傷中[19-22]，miR-877-5p表達上調(diào)且參與肝損傷過程。

注：-lg(P-value)：條形圖長度，表示富集指數(shù)。

圖2miR-877-5p靶基因富集的GO分析和KEGG通路分析

A：GO分類中分子功能領(lǐng)域(MF)；B：GO分類中細(xì)胞成分領(lǐng)域(CC)；C：GO分類中生物過程領(lǐng)域(BP)；D：KEGG通路分析結(jié)果

Fig2Top10significantfunctionalGOtermsandKEGGpathwaysofmiR-877-5ptargetgenes

A: the field of molecular function (MF) in GO classification; B: the field of cellular components (CC) in GO classification; C: the biological processes (BP) of GO classification; D: the analysis results of KEGG pathway

既往研究[23-29]發(fā)現(xiàn),瑞巴派特主要通過促進胃黏液的分泌，促進前列腺素E(postaglandin E2, PGE2)的產(chǎn)生，抑制炎癥細(xì)胞和炎癥因子的產(chǎn)生，拮抗氧化應(yīng)激等發(fā)揮胃黏膜保護作用。有研究[30-31]報道,瑞巴派特可通過改善胃腸道黏膜屏障來預(yù)防阿司匹林引起的胃黏膜損傷。但其是否通過改變miRNA發(fā)揮胃黏膜保護作用，目前尚無報道。

本研究發(fā)現(xiàn),阿司匹林損傷的胃黏膜上皮細(xì)胞株GES-1中miR-877-5p表達明顯升高，而經(jīng)瑞巴派特預(yù)處理后miR-877-5p的表達降低，說明miR-877-5p參與瑞巴派特保護阿司匹林所致胃黏膜損傷的過程。為進一步探討miR-877-5p的作用，我們向阿司匹林損傷組的GES-1細(xì)胞中轉(zhuǎn)染miR-877-5p inhibitor，發(fā)現(xiàn)細(xì)胞增殖能力增強，細(xì)胞凋亡減少；用miR-877-5p mimics轉(zhuǎn)染瑞巴派特保護組的GES-1細(xì)胞，細(xì)胞增殖受到抑制，凋亡增加，這些結(jié)果提示，miR-877-5p對GES-1細(xì)胞具有抑制增殖促進凋亡的作用。通過生物信息學(xué)分析發(fā)現(xiàn)，miR-877-5p可能參與細(xì)胞生長、增殖和凋亡等生物行為的調(diào)控過程。以上實驗說明，瑞巴派特通過抑制GES-1細(xì)胞中阿司匹林所誘導(dǎo)的miR-877-5p的表達，發(fā)揮胃黏膜保護作用。目前已知miRNA對細(xì)胞功能活動的影響往往是通過在轉(zhuǎn)錄后水平下調(diào)某些特定的功能基因，即靶基因的表達實現(xiàn)的[9]。本課題組下一步將在靶基因水平和蛋白水平上驗證miR-877-5p的靶基因在GES-1細(xì)胞中的功能，進一步證實miR-877-5p發(fā)揮的具體作用機制。

本研究證實了瑞巴派特發(fā)揮胃黏膜保護作用與miR-877-5p相關(guān)，通過抑制GES-1細(xì)胞中miR-877-5p的表達，減輕阿司匹林引起的胃黏膜上皮細(xì)胞的損傷。

[1] RAHME E, BERNATSKY S. NSAIDs and risk of lower gastrointestinal bleeding [J]. Lancet, 2010, 376(9736): 146-148. DOI: 10.1016/S0140-6736(10)60839-2.

[2] ZHANG S, QING Q, BAI Y, et al. Rebamipide helps defend against nonsteroidal anti-inflammatory drugs induced gastroenteropathy: a systematic review and meta-analysis [J]. Dig Dis Sci, 2013, 58(7): 1991-2000. DOI: 10.1007/s10620-013-2606-0.

[3] KIM H K, KIM J I, KIM J K, et al. Preventive effects of rebamipide on NSAID-induced gastric mucosal injury and reduction of gastric mucosal blood flow in healthy volunteers [J]. Dig Dis Sci, 2007, 52(8): 1776-1782. DOI: 10.1007/s10620-006-9367-y.

[4] KIM J H, PARK S H, CHO C S, et al. Preventive efficacy and safety of rebamipide in nonsteroidal anti-inflammatory drug-induced mucosal toxicity [J]. Gut Liver, 2014, 8(4): 371-379. DOI: 10.5009/gnl.2014.8.4.371.

[5] YEKTA S, SHIH I H, BARTEL D P. MicroRNA-directed cleavage of HOXB8 mRNA [J]. Science, 2004, 304(5670): 594-596. DOI: 10.1126/science.1097434.

[6] BARTEL D P. MicroRNAs: genomics, biogenesis, mechanism, and function [J]. Cell, 2004, 116(2):281-297.

[7] YU D, WU L, GILL P, et al. Multiple microRNAs function as self-protective modules in acetaminophen-induced hepatotoxicity in humans [J]. Arch Toxicol, 2018, 92(2): 845-858. DOI: 10.1007/s00204-017-2090-y.

[8] 孫沂, 樊宏偉, 王書奎, 等. 阿司匹林、氯吡格雷聯(lián)用對人胃黏膜上皮細(xì)胞株GES-1增殖的影響[J]. 胃腸病學(xué)和肝病學(xué)雜志, 2010, 19(6): 520-523.

SUN Y, FAN H W, WANG S K, et al. Effect of combined Aspirin and Clopidogrel on proliferation of human gastric epithelial cell line GES-1 [J]. Chin J Gastroenterol Hepatol, 2010, 19(6): 520-523.

[9] FRIEDMAN R C, FARH K K, BURGE C B, et al. Most mammalian mRNAs are conserved targets of microRNAs [J].Genome Res, 2009, 19(1): 92-105. DOI: 10.1101/gr.082701.

[10] KIM J, LEE K S, KIM J H, et al. Aspirin prevents TNF-α-induced endothelial cell dysfunction by regulating the NF-κB-dependent miR-155/eNOS pathway: Role of a miR-155/eNOS axis in preeclampsia [J]. Free Radic Biol Med, 2017, 104: 185-198. DOI: 10.1016/j.freeradbiomed.2017.01.010.

[11] YIANNAKOPOULOU E. Targeting epigenetic mechanisms and microRNAs by aspirin and other non-steroidal anti-inflammatory agents--implications for cancer treatment and chemoprevention [J]. Cell Oncol (Dordr), 2014, 37(3): 167-178. DOI: 10.1007/s13402-014-0175-7.

[12] LAN F, YUE X, HAN L, et al. Genome-wide identification of TCF7L2/TCF4 target miRNAs reveals a role for miR-21 in Wnt-driven epithelial cancer [J]. Int J Oncol, 2012, 40(2): 519-526. DOI: 10.3892/ijo.2011.1215.

[13] MIKAMI J, KUROKAWA Y, TAKAHASHI T, et al. Antitumor effect of antiplatelet agents in gastric cancer cells: an in vivo and in vitro study [J]. Gastric Cancer, 2016, 19(3): 817-826. DOI: 10.1007/s10120-015-0556-2.

[14] GUO X, YU L, CHEN M, et al. miR-145 mediated the role of aspirin in resisting VSMCs proliferation and anti-in ammation through CD40 [J]. J Transl Med, 2016, 14(1): 211. DOI: 10.1186/s12967-016-0961-2.

[15] WANG Y, GU X, LI Z, et al. microRNA expression profiling in multidrug resistance of the 5-Fu-induced SGC-7901 human gastric cancer cell line [J]. Mol Med Rep, 2013, 7(5): 1506-1510. DOI: 10.3892/mmr.2013.1384.

[16] SHI Q, XU X, LIU Q, et al. MicroRNA-877 acts as a tumor suppressor by directly targeting eEF2K in renal cell carcinoma [J]. Oncol Lett, 2016, 11(2): 1474-1480. DOI: 10.3892/ol.2015.4072.

[17] HUANG X, QIN J, LU S. Up-regulation of miR-877 induced by paclitaxel inhibits hepatocellular carcinoma cell proliferation though targeting FOXM1 [J]. Int J Clin Exp Pathol, 2015, 8(2): 1515-1524.

[18] LIANG Y, ZHAO G, TANG L, et al. MiR-100-3p and miR-877-3p regulate overproduction of IL-8 and IL-1β in mesangial cells activated by secretory IgA from IgA nephropathy patients [J]. Exp Cell Res, 2016, 347(2): 312-321. DOI: 10.1016/j.yexcr.2016.08.011.

[19] MITSUGI R, ITOH T, FUJIWARA R. MicroRNA-877-5p is involved in the trovafloxacin-induced liver injury [J]. Toxicol Lett, 2016, 263: 34-43. DOI: 10.1016/j.toxlet.2016.10.002.

[20] YAMASHITA Y, ASAKURA M, MITSUGI R, et al. microRNA expression in the vildagliptin-treated two- and three-dimensional HepG2 cells [J]. Drug Metab Pharmacokinet, 2016, 31(3): 201-209. DOI: 10.1016/j.dmpk.2016.02.004.

[21] YANNG X, GREENHAW J, SHI Q, et al. Identification of urinary microRNA profiles in rats that may diagnose hepatotoxicity [J]. Toxicol Sci, 2012, 125(2): 335-344. DOI: 10.1093/toxsci/kfr321.

[22] WARD J, BALA S, PETRASEK J, et al. Plasma microRNA profiles distinguish lethal injury in acetaminophen toxicity: a research study [J]. World J Gastroenterol, 2012, 18(22): 2798-2804. DOI: 10.3748/wjg.v18.i22.2798.

[23] ISHIHARA K, KOMURO Y, NISHIYAMA N, et al. Effect of rebamipide on mucus secretion by endogenous prostaglandin-independent mechanism in rat gastric mucosa [J]. Arzneimittelforschung, 1992, 42(12): 1462-1466.

[24] QI Z, JIE L, HAIXIA C, et al. Effect of rebamipide on quality of peptic ulcer healing in rat [J]. Dig Dis Sci, 2009, 54(9):1876-1883. DOI: 10.1007/s10620-008-0577-3.

[25] KLEINE A, KLUGE S, PESKAR B M. Stimulation of prostaglandin biosynthesis mediates gastroprotective effect of rebamipide in rats [J]. Dig Dis Sci, 1993, 38(8): 1441-1449.

[26] MURAKAMI K, OKAJIMA K, UCHIBA M, et al. Rebamipide attenuates indomethacin-induced gastric mucosal lesion formation by inhibiting activation of leukocytes in rats [J]. Dig Dis Sci, 1997, 42(2): 319-325.

[27] AIHARA M, AZUMA A, TAKIZAWA H, et al. Molecular analysis of suppression of interleukin-8 production by rebamipide in Helicobacter pylori-stimulated gastric cancer cell lines [J]. Dig Dis Sci, 1998, 43(9 Suppl): 174S-180S.

[28] YOSHIKAWA T, NAITO Y, TANIGAWA T, et al. Free radical scavenging activity of the novel anti-ulcer agent rebamipide studied by electron spin resonance [J]. Arzneimittelforschung, 1993, 43(3): 363-366.

[29] NAITO Y, YOSHIKAWA T, TANIGAWA T, et al. Hydroxyl radical scavenging by rebamipide and related compounds: electron paramagnetic resonance study [J]. Free Radic Biol Med, 1995, 18(1): 117-123.

[30] SUZUKI T, YOSHIDA N, NAKABE N, et al. Prophylactic effect of rebamipide on aspirin-induced gastric lesions and disruption of tight junctional protein zonula occludens-1 distribution [J]. J Pharmacol Sci, 2008, 106(3): 469-477.

[31] DIAO L, MEI Q, XU J M, et al. Rebamipide suppresses diclofenac-induced intestinal permeability via mitochondrial protection in mice [J]. World J Gastroenterol, 2012, 18(10): 1059-1066. DOI: 10.3748/wjg.v18.i10.1059.

doi:10.3969/j.issn.1006-5709.2018.06.016