Yamogenin是一种甾体皂苷,存在于芦笋等植物中,山药,foenum-graecumTrigonella,和Agavesp。在这项研究中,我们评估了体外细胞毒性,抗氧化剂,和羊草原蛋白的抗菌性能。对人类结肠癌HCT116,胃癌AGS,鳞癌UM-SCC-6细胞,和人正常成纤维细胞用MTT[3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物]测定。用流式细胞术估计用羊毛细胞素处理后的凋亡和死亡AGS细胞的量。此外,在Yamogenin处理的AGS细胞中,我们研究了活性氧(ROS)的产生,线粒体膜去极化,流式细胞术检测caspase-8和-9的活性水平和mRNA水平的基因表达,测光,和RT-PCR,分别。用DPPH(2,2-二苯基-1-吡啶酰肼)和ABTS(2,2'-叠氮基-双(3-乙基苯并噻唑啉-6-磺酸)测定法评估了拉莫宁的抗氧化性能。该化合物的抗菌潜力是在金黄色葡萄球菌上估计的,蜡样芽孢杆菌,肺炎克雷伯菌,大肠杆菌,肠沙门氏菌,幽门螺杆菌,弯曲杆菌,空肠弯曲杆菌,单核细胞增生李斯特菌,副干酪乳杆菌,和嗜酸乳杆菌菌株。在测试的细胞系中,Yamogenin对AGS细胞的细胞毒性作用最强(IC5018.50±1.24µg/mL)。与单一化合物相比,这种作用在拉马木素与奥沙利铂或卡培他滨的组合中显著更强。此外,产卵蛋白诱导的ROS产生,去极化的线粒体膜,并提高AGS细胞中caspase-8和-9的活性水平。RT-PCR分析显示,这种皂苷元在mRNA水平上强烈上调了TNFRSF25的表达。这些结果表明,产卵原蛋白通过外在和内在的凋亡方式诱导细胞死亡。抗氧化剂研究表明,羊草原蛋白具有中等的体外潜力(DPPH中的IC50为704.7±5.9µg/mL,ABTS测定中的IC50为631.09±3.51µg/mL)以及对蛋白质变性特性的抑制(IC50为1421.92±6.06µg/mL)。抗菌试验显示羊毛草素对细菌菌株的影响较弱,最强的是抗金黄色葡萄球菌(MIC值为350µg/mL)。总之,Yamogenin可能是治疗和预防胃癌的潜在候选者。
Yamogenin is a steroidal saponin occurring in plant species such as Asparagus officinalis, Dioscorea collettii, Trigonella foenum-graecum, and Agave sp. In this study, we evaluated in vitro cytotoxic, antioxidant, and antimicrobial properties of yamogenin. The cytotoxic activity was estimated on human colon cancer HCT116, gastric cancer AGS, squamous carcinoma UM-SCC-6 cells, and human normal fibroblasts with MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The amount of apoptotic and dead AGS cells after treatment with yamogenin was estimated with flow cytometry. Also, in yamogenin-treated AGS cells we investigated the reactive oxygen species (ROS) production, mitochondrial membrane depolarization, activity level of caspase-8 and -9, and gene expression at mRNA level with flow cytometry, luminometry, and RT-PCR, respectively. The antioxidant properties of yamogenin were assessed with DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2\'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assays. The antimicrobial potential of the compound was estimated on Staphylococcus aureus, Bacillus cereus, Klebsiella pneumoniae, Escherichia coli, Salmonella enterica, Helicobacter pylori, Campylobacter coli, Campylobacter jejuni, Listeria monocytogenes, Lactobacillus paracasei, and Lactobacillus acidophilus bacteria strains. Yamogenin showed the strongest cytotoxic effect on AGS cells (IC50 18.50 ± 1.24 µg/mL) among the tested cell lines. This effect was significantly stronger in combinations of yamogenin with oxaliplatin or capecitabine than for the single compounds. Furthermore, yamogenin induced ROS production, depolarized mitochondrial membrane, and increased the activity level of caspase-8 and -9 in AGS cells. RT-PCR analysis revealed that this sapogenin strongly up-regulated TNFRSF25 expression at the mRNA level. These results indicate that yamogenin induced cell death via the extrinsic and intrinsic way of apoptosis. Antioxidant study showed that yamogenin had moderate in vitro potential (IC50 704.7 ± 5.9 µg/mL in DPPH and 631.09 ± 3.51 µg/mL in ABTS assay) as well as the inhibition of protein denaturation properties (with IC50 1421.92 ± 6.06 µg/mL). Antimicrobial test revealed a weak effect of yamogenin on bacteria strains, the strongest one being against S. aureus (with MIC value of 350 µg/mL). In conclusion, yamogenin may be a potential candidate for the treatment and prevention of gastric cancers.