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目的:明确三种中药有效成分三七皂甙(PNS)、黄芪皂甙(As)及黄芩甙(Ba)单用或配伍应用对永生化的人胃黏膜上皮细胞GES-1以及经过甲基硝基亚硝基胍(MNNG) 转化后的GES-1细胞(MC细胞)增生能力的影响. 方法:以MNNG转化GES-1细胞,转化后细胞简称MC 细胞.分别以不同浓度的三七皂甙、黄芪皂甙及黄芩甙处理GES-1细胞及MC细胞,四甲基谷氮唑盐(MTT)法及软琼脂集落形成试验检测三种药物单用及配伍对细胞增生活性的影响.Annexin V及PI双染,流式细胞仪检测三药单用及配伍对MC细胞凋亡/坏死的影响;电镜和Hoechst33258及PI双染荧光显微镜观察凋亡细胞形态. 结果:PNS、AS及Ba对GES-1和MC细胞的增生活性有明显的抑制作用,并有随剂量增加作用增强的趋势.三药配伍应用对2种细胞的抑制作用增强(P<0.05或P<0.01vs三药单用组),还能明显抑制MC细胞的软琼脂集落形成能力(P<0.05或P<0.01 vs MC细胞对照组). 三药单用可引起MC细胞凋亡及死亡比例增加并呈一定的时间依赖性,配伍应用作用强于单独应用(P<0.05 或P<0.01 vs 三药单用组),增强程度从大至小依次为PNS+AS+Ba>PNS+AS>AS+Ba>PNS+Ba. 结论:三种中药有效成分单用及配伍可显著抑制MC细胞的增生并引起细胞死亡;引起细胞死亡的机制部分是通过诱导凋亡实现的;不同药物配伍应用的增效程度不同.
OBJECTIVE: To identify three active ingredients of Chinese medicine, PNS, Astragalus Saponins (As) and Astragalus (Ba), alone or in combination, for the immortalization of human gastric epithelial cells GES-1 and methylnitrosamine Effects of nitroguanidine (MNNG) on the proliferation of GES-1 cells (MC cells) transformed by nitroguanidine (MNNG). METHODS: Ginsenoside-1 (MGG-1) cells were transformed with MNNG and the transformed cells were abbreviated as MC cells. The concentrations of saponins and saponins were different. Treatment of GES-1 cells and MC cells with Astragalus membranaceus, MTT assay and soft agar colony formation assay were used to test the effects of three drugs alone and their compatibility on cell proliferation activity. Annexin V and PI double staining Flow cytometry was used to detect the effect of three drugs alone and compatibility on apoptosis/necrosis of MC cells; electron microscopy and Hoechst 33258 and PI dual staining fluorescence microscope were used to observe apoptotic cell morphology. Results: PNS, AS, and Ba for GES-1 and MC The proliferative activity of cells has a significant inhibitory effect, and there is a tendency to increase with the increase of dose. The inhibitory effect of the combination of three drugs on the two kinds of cells is enhanced (P<0.05 or P<0.01 vs three drugs alone group), but also Significantly inhibited soft agar colony formation of MC cells (P<0.05 or P<0.01 vs. MC cell control Group). Three drugs alone can cause apoptosis of MC cells and increase the proportion of death in a certain time-dependent manner, the compatibility of application is stronger than that of single use (P <0.05 or P <0.01 vs three drugs alone group), the degree of enhancement From the largest to the smallest, PNS+AS+Ba>PNS+AS>AS+Ba>PNS+Ba. Conclusion: The single use and compatibility of three active ingredients of Chinese herbs can significantly inhibit the proliferation of MC cells and cause cell death; causing cell death. The mechanism is partially achieved by inducing apoptosis; the degree of synergy of different drugs is different.