氯吡格雷对人胃黏膜上皮细胞紧密连结蛋白ZO-1表达的影响

目的 探讨氯吡格雷（Clopidogrel）对人胃黏膜上皮细胞（GES-1）的损伤机制.方法 建立GES-1单层细胞模型,将细胞分为阴性对照组、U0126干预组、氯吡格雷干预组、U0126预处理后氯吡格雷干预组（联合组）,采用MTT比色法和流式细胞术检测各组细胞增殖、凋亡情况;免疫细胞化学检测各组p-ERK1/2的表达情况;采用Western blotting检测各细胞组p-ERK1/2和紧密连接蛋白ZO-1的表达量.结果 与阴性对照组比较,U0126干预组、氯吡格雷干预组、U0126预处理后氯吡格雷干预组的细胞增殖明显受到抑制（P＜0.05）;Western blotting结果显示：与阴性对照组比较,后3组的p-ERK1/2表达下降,与免疫细胞化学的趋势一致;ZO-1表达趋势亦与p-ERK1/2表达一致.结论 在GES-1细胞模型中,氯吡格雷可能通过抑制p-ERK1/2的表达来降低ZO-1的表达,从而损伤GES-1细胞.

Effect of Clopidogrel on the tight junctional ZO-1 of human gastric epithelial cell

Objective To investigate the damage mechanism of Clopidogrel in human gastric epithelial GES-1 cells.Methods GES-1 cells were cultured in vitro.Then the GES-1 cells were divided into four groups：control group,U0126 group,Clopidogrel group and U0126 ＋ Clopidogrel group,the reduced proliferation rates and apoptosis rates of GES-1 cells in four groups were examined by methyl thiazolyl tetrazolium （MTT） assay and Flow cytometry.The expression of phosphorylated ERK1/2 protein in GES-1 cells was detected by inmunohistochemistry method and the expression of phosphorylated ERK1/2 protein and ZO-1 protein in GES-1 cells were detected by Western blotting.Results The result of MTT showed that the proliferation of GES-1 cells was inhibited in other three groups compared with control group （P ＜ 0.05） ; Flow cytometry analysis indicated that the apoptosis rate of cells in U0126,Clopidogrel and the U0126 ＋Clopidogrel groups was higher than that in control group （P ＜ 0.05）.The expression of p-ERK1/2 and ZO-1 proteins in other three groups were lower than those in control group （P ＜ 0.01）.Conclusion The injury mechanism of Clopidogrel on human gastric epithelial GES-1 cells may inhibit the expression of p-ERK1/2 to decrease the expression of ZO-1.