瑞巴派特对氯吡格雷所致人胃黏膜上皮细胞损伤的保护作用及其机制研究

Protective Effect and Mechanism of Rebamipide on Human Gastric Mucosal Epithelial Cell Injury Induced by Clopidogrel

背景:长期服用氯吡格雷可引起胃肠道损伤。瑞巴派特是一种新型胃黏膜保护剂,其用于防治氯吡格雷相关胃黏膜损伤的疗效尚不明确。目的:探讨瑞巴派特对氯吡格雷所致人胃黏膜上皮细胞损伤的保护作用及其可能机制。方法:以氯吡格雷处理的人胃黏膜上皮细胞株GES-1作为氯吡格雷组,以氯吡格雷联合不同浓度(0.2、0.5、1.0 mmol/L)瑞巴派特预处理的GES-1细胞作为瑞巴派特组,同时设立阴性对照组。采用MTT法检测细胞增殖抑制情况;采用倒置相差显微镜观察细胞形态学变化;采用蛋白质印迹法检测三叶因子1(TFF1)、磷酸化细胞外信号调节激酶1/2(p-ERK1/2)表达。结果:MTT法检测结果显示,瑞巴派特0.2、0.5、1.0 mmol/L组GES-1细胞增殖抑制率均显著低于氯吡格雷组(P<0.05)。倒置相差显微镜观察发现,氯吡格雷可诱导GES-1细胞损伤,而瑞巴派特可减轻氯吡格雷所致的细胞损伤。蛋白质印迹法检测结果显示,氯吡格雷组GES-1细胞TFF1蛋白表达较阴性对照组显著升高(P<0.05);瑞巴派特0.2、0.5、1.0 mmol/L组TFF1蛋白表达均较氯吡格雷组进一步升高,且作用呈浓度依赖性(P<0.05)。氯吡格雷组GES-1细胞p-ERK1/2蛋白表达较阴性对照组显著升高(P<0.05);瑞巴派特0.2、0.5、1.0 mmol/L组p-ERK1/2蛋白表达均较氯吡格雷组显著降低,作用亦呈浓度依赖性(P<0.05)。结论:瑞巴派特可能通过抑制ERK信号通路上调TFF1蛋白表达,从而参与调控氯吡格雷所致GES-1细胞损伤的修复,此过程是瑞巴派特防治氯吡格雷相关胃黏膜损伤的可能机制之一。

Long-term use of elopidogrel may cause gastrointestinal injury. Rebamipide is a novel mucoprotective drug, however, its protective effect on gastric mucosal injury induced by clopidogrel has not been clarified. Aims： To investigate the protective effect and potential mechanism of rebamipide on human gastric mueosal epithelial cell injury induced by clopidogrel. Methods： In clopidogrel group, human gastric mucosal epithelial cell line GES-1 was treated with elopidogrel; GES-1 ceils in rebamipide group were pretreated with different concentrations of rebamipide （0.2, O. 5 and 1.0 mmol/L） before clopidogrel treatment, and a negative control group was established simultaneously. Inhibition of cell proliferation was measured by MTF assay. Morphological change of cell was observed by inverted phase contrast microscope. Expressions of trefoil factor family 1 （TFF1） and phosphorylated extracellular signal regulated kinase 1/2 （ p- ERK1/2） were determined by Western blotting. Results： Mq^F assay showed that cell proliferation inhibition rates of GES-1 cells in rebamipide 0.2, 0.5 and 1.0 mmol/L groups were significantly lower than that in clopidogrel group （ P 〈 0.05）. Injury of GES-1 cells was seen in clopidogrel group by inverted phase contrast microscope and rebamipide could reduce the cell injury. Expression of TFF1 in GES-1 cells in clopidogrel group was significantly higher than that in negative control group （P 〈0.05）, while those in rebamipide 0.2, 0.5 and 1.0 mmol/L groups were further increased in a dose-dependent manner as compared with that in clopidogrel group （P 〈 0.05）. Expression of p-ERK1/2 in GES-1 cells in clopidogrel group was significantly higher than that in negative control group （P 〈 0.05）, while those in rebamipide 0.2, 0.5 and I. 0 mmol/L groups were significantly lower than that in clopidogrel group in a dose-dependent manner （ P 〈 0. 05 ）. Conclusions ：Rebamipide has a protective effect on GES-1 cells injury induced by clopidogrel, probably through inhibiting ERK signaling pathway, up-regulating TFF1 expression and promoting mucosal healing.