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目的肠三叶因子(intestinal trefoil factor,ITF)对胃黏膜具有保护作用,但其机制尚不明确。文中探讨重组ITF对胃黏膜上皮细胞增殖能力的影响及其可能作用的分子机制。方法将体外培养的GES-1细胞分为3组,其中1组作为细胞增殖活力实验空白对照组,其他2组分别加入浓度为100和500 ng/m L的人重组ITF形成低浓度ITF组和高浓度ITF组。光镜下观察细胞形态,并采用CCK-8试剂盒检测GES-1细胞的增殖活力。体外培养的GES-1细胞株分别用100 ng/m L的ITF和浓度为15μmol/L的PI3K/Akt信号通路抑制剂LY294002进行处理,分为信号通路蛋白表达实验空白对照组、ITF组、LY294002组和ITF+LY294002组,光镜下观察细胞形态。随后采用CCK-8试剂盒检测GES-1细胞处理后的增殖活力,采用Western blot检测PI3K/Akt信号通路中p-Akt和Akt蛋白的表达情况。结果与空白对照组相比,ITF刺激下GES-1细胞增殖活力明显增强,且ITF浓度越高、增殖活力越强。使用LY294002处理后,能够显著抑制ITF刺激的细胞增殖。Western blot检测结果表明,ITF提高了p-Akt蛋白的表达水平,激活PI3K/Akt信号通路。结论 ITF促进GES-1细胞的增殖,推测其分子作用机制主要是通过激活PI3K/Akt信号通路促进细胞增殖。
Purpose Intestinal trefoil factor (ITF) has a protective effect on gastric mucosa, but its mechanism is not yet clear. The article discusses the effect of recombinant ITF on the proliferation of gastric mucosal epithelial cells and its molecular mechanism. Methods GES-1 cells cultured in vitro were divided into three groups. One group served as blank control group, and the other two groups were added with ITF group with low concentration and ITF group with 100 and 500 ng / mL respectively High-concentration ITF group. Cell morphology was observed under light microscope, and the viability of GES-1 cells was detected by CCK-8 kit. The cultured GES-1 cell lines were treated with 100 ng / mL ITF and 15 μmol / L PI3K / Akt signaling pathway inhibitor LY294002, respectively, and divided into control group, ITF group, LY294002 Group and ITF + LY294002 group, the cell morphology was observed under light microscope. Subsequently, the proliferation activity of GES-1 cells was detected by CCK-8 kit. The expression of p-Akt and Akt in PI3K / Akt signaling pathway was detected by Western blot. Results Compared with the blank control group, the proliferative activity of GES-1 cells was significantly increased under the stimulation of ITF, and the higher the ITF concentration, the stronger the proliferative activity. Treatment with LY294002 significantly inhibited ITF-stimulated cell proliferation. Western blot results showed that ITF increased the expression of p-Akt protein and activated the PI3K / Akt signaling pathway. Conclusion ITF can promote the proliferation of GES-1 cells. It is speculated that the molecular mechanism of ITF is to promote cell proliferation by activating the PI3K / Akt signaling pathway.