杭白菊中黄酮类化合物及其抑制人源肿瘤细胞增殖活性的研究

 目的 分离和鉴定杭白菊中黄酮类化合物并对其进行抗肿瘤活性研究。方法 利用系统溶剂萃取法、聚酰胺柱色谱、半制备型高效液相色谱法分离。通过理化性质和IR、UV、MS、¹H-NMR、¹³C-NMR等光谱数据鉴定化合物。采用MTT法测定化合物对人肿瘤细胞增殖的影响, 并以氟尿嘧啶（5-FU）作为阳性对照,比较其对正常细胞的毒性作用。流式细胞术检测细胞周期分布的改变。结果 分离得到8个黄酮化合物单体,分别鉴定为木犀草素(Ⅰ),木犀草素7-O-β-D-葡萄糖苷(Ⅱ),木犀草素7-O-β-D (6″-O-丙二酸单酰) -葡萄糖苷(Ⅲ),芹菜素(Ⅳ),芹菜素 7-O-β-D-葡萄糖苷(Ⅴ),芹菜素 7-O-β-D-(6″-O-丙二酸单酰)-葡萄糖苷(Ⅵ),香叶木素(Ⅶ),香叶木素 7-O-β-D-葡萄糖苷(Ⅷ)。MTT法测定木犀草素抑制乳腺癌MDA-MB-231细胞 和肝癌SMMC-7721细胞的IC₅₀分别为56.67和72.92 μmol·L^-1。结论 化合物Ⅲ和Ⅵ首次从杭白菊中分离得到。木犀草素能抑制MDA-MB-231和SMMC-7721增殖,阻滞其细胞周期于G2/M期,且对正常细胞无毒性。

Evaluation of Flavonoids from Flos chrysanthemi on Cell Proliferation in Human Cancer Cells

OBJECTIVE To isolate and identify flavonoids from Flos chrysanthemi and to study the anticancer bioactivity. METHODS Compounds were isolated by systematic solvents and purified by chromatography of polyamide column and Semi-preparative HPLC. Their structures were identified by physical and chemical properties and variety of spectrum data, including IR, UV, MS, ¹H-NMR and ¹³C-NMR. The anti-proliferation effect of theses compounds on human breast cancer and liver cancer cells was studied by MTT assay, comparing the compound cytotoxicity on normal tissue with 5-fluorouracil（5-Fu） as the positive control. Cell cycle change was detected using propidium iodide staining by flow cytometry . RESULTS Eight compounds were obtained and identified as luteolin(Ⅰ), luteolin 7-O-β-D-glucoside(Ⅱ), luteolin 7-O-β-D-(6″-O-malonyl)-glucoside (Ⅲ), apigenin (Ⅳ), apigenin7-O-β-D-glucoside(Ⅴ), apigenin 7-O-β-D-(6″-O-malonyl)-glucoside)(Ⅵ), diosmetin (Ⅶ), and diosmetin 7-O-β-D-glucoside (Ⅷ), respectively. MTT assay demonstrated that luteolin inhibited proliferation of both the MDA-MB-231 (IC₅₀=56.67 μmol·L^-1) and SMMC-7721 (IC₅₀=72.92 μmol·L^-1) cell lines. CONCLUSION Compound Ⅲ and Ⅵ were obtained from Flos chrysanthemi for the first time. Luteolin inhibited the proliferation of both the MDA-MB-231 and SMMC-7721 and arrested cell cycle at G2/M phase, without cytotoxicity toward normal cells.