瑞巴派特对NSAIDs小肠损伤的保护作用及机制研究
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摘要
目的:探讨瑞巴派特对NSAIDs相关小肠损伤的保护作用,分析该保护机制与TLR4/NF-κB信号通路的关系。方法:采用双氯芬酸制作NSAIDsSD大鼠小肠黏膜损伤模型,瑞巴派特进行干预。对肠黏膜组织行大体形态学观察、组织病理学评分,ELISA、免疫组化、免疫印迹及PCR检测TLR4/NF-κB等相关蛋白在肠黏膜中的表达情况。结果:瑞巴派特干预组小肠黏膜组织的大体评分及组织病理学评分明显低于模型组(P<0.01);细胞紧密连接蛋白Claudin-1及ZO-1的表达在模型组表达下调,在瑞巴派特干预组明显高于模型组(P<0.01)。TLR4及NF-κBp65蛋白及IL-1β、IL-8、TNF-α含量在模型组小肠黏膜中表达升高,经瑞巴派特干预后表达下降(P<0.05);而负性调控因子Tollip蛋白在模型组表达降低,经瑞巴派特干预后Tollip的表达明显上调(P<0.01)。PPAR-γ表达在3组表达无明显差异(P>0.05)结论:瑞巴派特有效降低了肠道黏膜的炎症反应,可能与抑制TLR4/NF-κBp65信号通路的过度激活有关。
OBJECTIVE To investigate the protective effect of rebamipide on NSAIDs-associated intestinal injury and analyze the relationship between the protective mechanism and TLR4/NF-κB signaling pathway.METHODS The small intestinal mucosal injury model of NSAIDs SD rats was established by diclofenac, and rebamipide was used for intervention. The expression of TLR4/NF-κB and other related proteins in intestinal mucosa was detected by ELISA, immunohistochemistry, Western blotting and PCR. RESULTS In rebamipide intervention group, the scores of gross and histopathology of small intestinal mucosa were significantly lower than those in model group(P<0.01). The expressions of tight junction proteins claudin-1 and ZO-1 were down-regulated in model group, which were significantly higher in rebamipide intervention group than those in model group(P<0.01). The expression of TLR4 and NF-κBp65 protein and the expression of IL-1β, IL-8 and TNF-α in the small intestinal mucosa of the model group was up-regulated, and then decreased after intervention with rebamipide(P<0.05). Howeverr, the expression of the negative regulator Tollip protein decreased in the model group and was significantly up-regulated after intervention with rebamipide(P<0.01). There was no significant difference in PPAR-expression among the three groups(P>0.05). CONCLUSION Rebamipide effectively reduces NSAIDs-mediated inflammation of intestinal mucosa probably by suppressing the over-activation of TLR4/NF-κB signaling pathway.
OBJECTIVE To investigate the protective effect of rebamipide on NSAIDs-associated intestinal injury and analyze the relationship between the protective mechanism and TLR4/NF-κB signaling pathway.METHODS The small intestinal mucosal injury model of NSAIDs SD rats was established by diclofenac, and rebamipide was used for intervention. The expression of TLR4/NF-κB and other related proteins in intestinal mucosa was detected by ELISA, immunohistochemistry, Western blotting and PCR. RESULTS In rebamipide intervention group, the scores of gross and histopathology of small intestinal mucosa were significantly lower than those in model group(P<0.01). The expressions of tight junction proteins claudin-1 and ZO-1 were down-regulated in model group, which were significantly higher in rebamipide intervention group than those in model group(P<0.01). The expression of TLR4 and NF-κBp65 protein and the expression of IL-1β, IL-8 and TNF-α in the small intestinal mucosa of the model group was up-regulated, and then decreased after intervention with rebamipide(P<0.05). Howeverr, the expression of the negative regulator Tollip protein decreased in the model group and was significantly up-regulated after intervention with rebamipide(P<0.01). There was no significant difference in PPAR-expression among the three groups(P>0.05). CONCLUSION Rebamipide effectively reduces NSAIDs-mediated inflammation of intestinal mucosa probably by suppressing the over-activation of TLR4/NF-κB signaling pathway.
引文
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[18] Imaeda H,Fujimoto T,Takahashi K,et al.Terminal-restriction fragment length polymorphism (T-RFLP) analysis for changes in the gut microbiota profiles of indomethacin-and rebamipide-treated mice[J].Digestion,2012,86(3):250-257.
[19] Endo H,Higurashi T,Hosono K,et al.Efficacy of lactobacillus casei treatment on small bowel injury in chronic low-dose aspirin users:a pilot randomized controlled study[J].J Gastroenterol,2011,46(7):894-905.
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[2] Jaafar MH,Safi SZ,Tan MP,et al.Efficacy of Rebamipide in Organic and Functional Dyspepsia:A Systematic Review and Meta-Analysis[J].Dig Dis Sci,2018,63(5):1250-1260.
[3] Fujimoto A,Uraoka T,Nishizawa T,et al.Rebamipide solution:a novel submucosal injection material to promote healing speed and healing quality of ulcers induced by endoscopic submucosal dissection[J].Gastrointest Endosc,2018,87(4):1114-1120.
[4] Fujimori S,Takahashi Y,Gudis K,et al.Rebamipide has the potential to reduce the intensity of NSAID-induced small intestinal injury:a double-blind,randomized,controlled trial evaluated by capsule endoscopy[J].J Gastroenterol,2011,46(1):57-64.
[5] Tomita T,Sadakata H,Tamura M,et al.Indomethacin-induced generation of reactive oxygen species leads to epithelial cell injury before the formation of intestinal lesions in mice[J].J Physiol Pharmacol,2014,65(3):435-440.
[6] Tanigawa T,Watanabe T,Otani K,et al.Rebamipide inhibits indomethacin-induced small intestinal injury:possible involvement of intestinal microbiota modulation by upregulation of α-defensin 5[J].Eur J Pharmacol,2013,704(1-3):64-69.
[7] Yamada S,Naito Y,Takagi T,et al.Rebamipide ameliorates indomethacin-induced small intestinal injury in rats via the inhibition of matrix metallo proteinases activity[J].J Gastroenterol Hepatol,2012,27(12):1816-1824.
[8] Lai Y,Zhong W,Yu T,et al.Rebamipide promotes the regeneration of aspirin-induced small-intestine mucosal injury through accumulation of β-Catenin[J].PLoS One,2015,10(7):e0132031.
[9] Yang C,Cui MH,Liang J,et al.Protective effect of Rebamipide on the small intestinal permeability in rats with Aspirin-induced[J].Chin J Gastroenterol Hepatol,2017,26(11):1258-1262.
[10] Liu F,Koval M,Ranganathan S,et al.Systems Proteomics View of the Endogenous Human Claudin Protein Family[J].J Proteome Res,2016,15(2):339-359.
[11] Chelakkot C,Ghim J,Ryu SH.Mechanisms regulating intestinal barrier integrity and its pathological implications[J].Exp Mol Med,2018,50(8):103.
[12] Zhang S,Zheng S,Wang X,et al.Carbon monoxide-releasing molecule-2 reduces intestinal epithelial tight-junction damage and mortality in septic rats[J].PLoS One,2015,10(12):e145988.
[13] Takeuchi K,Satoh H.NSAID-induced small intestinal damage- roles of various pathogenic factors[J].Digestion,2015,91:218-232.
[14] Colucci R,Pellegrini C,Fornai M,et al.Pathophysiology of NSAID-Associated Intestinal Lesions in the Rat:Luminal Bacteria and Mucosal Inflammation as Targets for Prevention[J].Front Pharmacol,2018,9:1340.
[15] Otani K,Tanigawa T,Watanabe T,et al.Microbiota Plays a Key Role in Non-Steroidal Anti-Inflammatory Drug-Induced Small Intestinal Damage[J].Digestion,2017,95(1):22-28.
[16] Nadatani Y,Watanabe T,Tanigawa T,et al.High mobility group box 1 promotes small intestinal damage induced by nonsteroidal anti-inflammatory drugs through toll-like receptor 4[J].Am J Pathol 2012,181(1):98-110.
[17] Higashimori A,Watanabe T,Nadatani Y,et al.Mechanisms of NLRP3 inflammasome activation and its role in NSAID-induced enteropathy[J].Mucosal Immunol,2016,9(3):659-668.
[18] Imaeda H,Fujimoto T,Takahashi K,et al.Terminal-restriction fragment length polymorphism (T-RFLP) analysis for changes in the gut microbiota profiles of indomethacin-and rebamipide-treated mice[J].Digestion,2012,86(3):250-257.
[19] Endo H,Higurashi T,Hosono K,et al.Efficacy of lactobacillus casei treatment on small bowel injury in chronic low-dose aspirin users:a pilot randomized controlled study[J].J Gastroenterol,2011,46(7):894-905.
[20] Dai C,Zheng CQ,Meng FJ,et al.VSL#3 probiotics exerts the anti-inflammatory activity via PI3k/Akt and NF-κB pathway in rat model of DSS-induced colitis[J].Mol Cell Biochem,2013,374(1-2):1-11.