莲房原花青素对肿瘤细胞及正常细胞毒作用的比较

Procyanidins from lotus seedpod and their cytotoxity against cancer cell lines as well as normal cells

目的 :研究莲房原花青素总提取物LSPC的含量和相对分子质量分布及其对人口腔表皮样癌 (humannasopharyngealepidermoidcarci noma ,KB)细胞、大鼠嗜碱粒细胞 (ratbasophilicleukemiacell ,RBL 2H3 )、小鼠成纤维细胞 (mousefibrosarcomacell ,L92 9)和人肥大细胞 (humanmastcellline ,HMC 1)的毒作用。方法 :用ESI MS分析LSPC ;并用倒置显微镜观察细胞形态 ,MTT法检测细胞增殖和流式细胞术检测细胞周期和细胞凋亡 ,研究LSPC对KB、RBL 2H3、L92 9和HMC 1细胞的毒作用。结果 :LSPC含量 >98% ,LSPC含单体、二聚体、三聚体、四聚体原花青素和原花青素二聚体酸单酯和三聚体酸单酯 ,相对分子质量为 2 90～ 115 4 3。当LSPC浓度为 10 0～ 40 0 μg/mL时 ,倒置显微镜下可见KB和RBL 2H3细胞变圆、皱缩、漂浮 ,而L92 9和HMC 1细胞变化不明显。MTT法实验表明 ,LSPC对KB和RBL 2H3细胞的生长呈显著的抑制作用 ,而对L92 9和HMC 1细胞抑制作用不明显 ;流式细胞术细胞周期和细胞凋亡分析进一步证明 ,LSPC可阻止KB和RBL 2H3细胞增殖并诱导其凋亡。结论 :LSPC对肿瘤细胞具有较强的毒作用 ,而对正常细胞毒性较小。

OBJECTIVE:To study the contents and constituents of LSPC (procyanidins from lotus seedpod) and its cytotoxity against human nasopharyngeal epidermoid carcinoma (KB) cells, rat basophilic leukemia cells (RBL-2H3), mouse fibrosarcoma cells (L929) and human mast cell line (HMC-1). METHODS: ESI-MS was used to analyze LSPC, whose effects on KB, RBL-2H3, L929 and HMC-1 cells were studied by upside-down microscopy, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay,and the cell cycle and cell apoptosis analysis. RESULTS: The contents of LSPC were exceeded 98% and mono-, di-, tri-, tetrameric procyanidins and di-, trimeric galic ester were constituted of LSPC with the relative molecular weight distribution ranging from 290- 1 154.3 . When the concentration of LSPC was 100-400 μg/mL, the morphology of KB and RBL-2H3 cells changed to be round, folded and the cells even were floating but the morphological changes of L929 and HMC-1 cells was little under the upside-down microscope. The MTT assay demonstrated that LSPC inhabited the growth of KB and RBL-2H3 cells rather than L929 and HMC-1 cells. Furthermore, the results of cell cycle and cell apoptosis analysis showed that LSPC hampered the proliferation of KB and RBL-2H3 cells and induced their apoptosis.CONCLUSION: LSPC has stronger cytotoxity against cancer cell lines than normal cells.