目的 探讨橙皮苷对胃癌AGS细胞的诱导凋亡作用及其相关分子机制。方法 采用MTT比色法检测橙皮苷对AGS细胞的杀伤作用；Annexin V-FITC/PI双染法和流式细胞术检测橙皮苷对AGS细胞的诱导凋亡作用、活性氧水平及加入活性氧清除剂N-乙酰-L-半胱氨酸（NAC）后细胞凋亡变化情况；Western blotting法检测细胞凋亡相关蛋白和丝裂原活化蛋白激酶（MAPK）信号通路相关蛋白的表达。结果 MTT比色法检测结果显示橙皮苷对AGS具有良好的增殖抑制作用。经橙皮苷处理后AGS细胞呈现细胞核固缩、细胞皱缩等凋亡现象。Annexin V-FITC/PI双染法和流式细胞术检测结果表明橙皮苷可以诱导AGS细胞发生线粒体依赖性凋亡，增加细胞内活性氧的水平，预处理NAC后，橙皮苷的诱导凋亡作用被抑制。Western blotting检测结果显示p-JNK、p-p38、Bad、cleaved Caspase-3及活化的聚腺苷二磷酸核糖聚合酶（cleave PARP）蛋白表达水平升高，抗凋亡蛋白磷酸化细胞外调节蛋白激酶（p-ERK）及Bcl-2表达水平降低，说明橙皮苷激活了AGS细胞中的MAPK信号通路及线粒体依赖性凋亡。结论 橙皮苷对AGS细胞具有良好的杀伤作用，并通过提高AGS细胞内活性氧水平，进而调控MAPK信号通路来诱导AGS细胞发生线粒体依赖性凋亡。

Objective To investigate the effect of hesperidin on apoptosis of gastric cancer AGS cells and its related molecular mechanisms. Methods MTT assay was used for the killing effect of hesperidin on human gastric cancer AGS cells; Annexin V-FITC/PI double staining and flow cytometry was used to detect the apoptosis induced by hesperidin on AGS cells, the level of reactive oxygen species, and the addition of NAC Post-apoptosis changes; Western blotting was used to detect the expression of apoptosis-related proteins and signaling pathway-related proteins. Results MTT assay showed that hesperidin had a good inhibiting effect on AGS cells. After treated with hesperidin, AGS cells showed apoptosis such as nuclear condensation and cell shrinkage. Annexin V-FITC/PI double staining and flow cytometry showed that hesperidin can induce mitochondrial dependent apoptosis of AGS cells and increase the level of intracellular reactive oxygen species. After pretreatment of NAC, hesperidin induced apoptosis inhibition. The results of Western blotting showed that the expression of p-JNK, p-p38, Bad, cleaved Caspase-3, and cleaved PARP increased, and the expression of anti-apoptotic proteins p-ERK and Bcl-2 decreased, which indicated that hesperidin activated the MAPK signaling pathway and mitochondria-dependent apoptosis in AGS cells. Conclusion Hesperidin has a good killing effect on human gastric cancer AGS cells, and induces mitochondria-dependent apoptosis in AGS cells by increasing the level of reactive oxygen species in AGS cells and regulating MAPK signaling pathway.

宁夏自然科学基金：转录因子OCT1调控ERK信号通路对胃癌细胞增殖侵袭的影响及机制研究（NZ16132）