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为了获得具有显著抑瘤活性的水溶性海参皂苷样品,利用大孔树脂柱、硅胶柱和葡聚糖凝胶柱对水溶性海参皂苷进行了分离纯化,并对其体内外抑瘤活性和诱导肿瘤细胞凋亡进行了检测。研究结果显示,大孔树脂柱的70%乙醇洗脱组分以及硅胶柱色谱的部分纯化组分(pSC-2和pSC-3)对HeLa细胞、A-549肺癌细胞、SGC-7901胃癌细胞和Bel-7402肝癌细胞均具有显著的抑瘤活性;经过葡聚糖凝胶柱色谱,获得了纯度均在99.6%以上、具有三萜类皂苷性质的SC-2和SC-3纯化样品,发现SC-2纯化样品不仅能浓度和时间依赖性地抑制HeLa细胞的体外增殖,还能剂量依赖性地抑制S180实体瘤的体内生长,提高荷瘤小鼠的胸腺指数和脾指数。细胞凋亡检测结果显示,SC-2纯化样品可剂量依赖性地诱导HeLa细胞凋亡,SC-2纯化样品(10和50mg.L-1)处理12h的HeLa细胞均出现了DNA片段化。可见,水溶性海参皂苷SC-2纯化样品具有显著的抑瘤活性,其机制可能是通过诱导肿瘤细胞发生凋亡来实现的。
In order to obtain water-soluble sea cucumber saponin samples with significant antitumor activity, the water-soluble sea cucumber saponins were isolated and purified by macroporous resin column, silica gel column and Sephadex G-30 column, and their antitumor activity and tumor induction in vitro and in vivo Apoptosis was detected. The results showed that 70% ethanol elution of macroporous resin column and partial purification of silica gel column (pSC-2 and pSC-3) on HeLa cells, A-549 lung cancer cells, SGC-7901 gastric cancer cells and Bel-7402 hepatocarcinoma cells had significant antitumor activity. The purified samples of SC-2 and SC-3 with purity of above 99.6% were obtained by Sephadex column. The results showed that SC -2 purified samples not only inhibited the proliferation of HeLa cells in vitro but also inhibited the growth of S180 solid tumors in vivo and the thymus index and spleen index of tumor-bearing mice in a dose-dependent manner. The results of apoptosis assay showed that SC-2 purified cells induced apoptosis in HeLa cells in a dose-dependent manner. DNA fragmentation occurred in HeLa cells treated with 10 and 50 mg.L-1 SC-2 for 12 hours. It can be seen that the purified sample of water-soluble sea cucumber saponin SC-2 has significant anti-tumor activity, and its mechanism may be achieved by inducing tumor cell apoptosis.