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山东大学学报(医学版) ›› 2017, Vol. 55 ›› Issue (8): 35-41.doi: 10.6040/j.issn.1671-7554.0.2017.070

• 基础医学 • 上一篇    下一篇

槲寄生中酚类化学成分的分离鉴定及其对A549细胞的增殖抑制活性

黄竹青,吴雪韦,任冬梅   

  1. 山东大学药学院 天然产物化学生物学教育部重点实验室, 山东 济南 250012
  • 收稿日期:2017-01-18 出版日期:2017-08-10 发布日期:2017-08-10
  • 通讯作者: 任冬梅. E-mail:rendom@sdu.edu.cn E-mail:rendom@sdu.edu.cn
  • 基金资助:
    山东省自然科学基金(2015ZRE27209)

Isolation and identification of phenolic constituents from Viscum coloratum and the antiproliferative effects on A549 cells

HUANG Zhuqing, WU Xuewei, REN Dongmei   

  1. Key Lab of Chemical Biology(Ministry of Education), School of Pharmaceutical Sciences, Shandong University, Jinan 250012, Shandong, China
  • Received:2017-01-18 Online:2017-08-10 Published:2017-08-10

摘要: 目的 分离纯化槲寄生抗肺癌A549细胞增殖活性的化学成分,对获得的活性成分促A549细胞凋亡的作用进行评价。 方法 采用硅胶柱层析、Sephadex LH-20凝胶柱层析等方法对槲寄生提取物的乙酸乙酯萃取部分进行分离纯化;1H NMR和13C NMR波谱法确定化合物结构;MTT法检测化合物对细胞增殖的影响;流式细胞术检测细胞周期、凋亡及胞内活性氧(ROS)的变化;蛋白免疫印迹法检测蛋白表达的变化。 结果 获得了10个酚类化合物;其中化合物1为1,7-二(4-羟基苯基)-4E,6E-庚二烯-3-酮,对A549细胞表现出剂量依赖的增殖抑制作用(F=196.768, P=0.001),可引起G2/M期细胞增多(P<0.05),S期细胞减少(P<0.05);化合物1可引起胞内ROS水平显著升高(P<0.05),促进细胞凋亡,并引起细胞Caspase-9,Caspase-3和PARP蛋白表达水平的改变(F值分别为19.340、4.388、194.571,P值分别为0.001、0.042、0.001)。 结论 槲寄生中的酚类成分主要是二苯基庚烷类和黄酮类,二苯基庚烷类化合物是该药材抗肺癌细胞增殖的主要活性成分。

关键词: 肺肿瘤, A549细胞, 1, 槲寄生, 6E-庚二烯-3-酮, 7-二(4-羟基苯基)-4E

Abstract: Objective To isolate and purify the chemical constituents of the active fraction from Viscum coloratum, and to evaluate the apoptotic effects on A549 cells. Methods Various chromatographic methods including silica gel, sephadex LH-20 column chromatography were used for the purification of chemical constituents from the extracts of Viscum coloratum. Spectroscopic methods including NMR and MS were used for the structural elucidation of the compounds. The antiproliferative effects were evaluated by MTT assay. Flow cytometry was used for the determination of cell cycle, apoptosis and intracellular reactive oxygen species(ROS)level. Western blotting analysis was carried out for the detection of protein expression. Results Ten phenolic compounds were obtained from Viscum coloratum. Compound 1 was identified as 1,7-di-(4-hydroxyphenyl)-4E,6E-heptadiene-3-ketone. This compound exerted cytotoxicity on A549 cells dose-dependently(F=196.768, P=0.001). It increased the cell number in G2/M phase(P<0.05)and decreased the cell number in S phase(P<0.05). The intracellular ROS level induced by compound 1 increased significantly(P<0.05). The compound also induced apoptosis of A549 cells. The expressions of Caspase-9, Caspase-3 and PARP were changed under the treatment(F: 19.340, 4.388, 194.571; P: 0.001, 0.042, 0.001). Conclusion Diphenylheptanes and flavonoids are the main phenolic components of Viscum coloratum, and diphenylheptanes are the main active ingredients of Viscum coloratum against lung cancer cell proliferation.

Key words: 1,7-di-(4-hydroxyphenyl)-4E,6E-heptadiene-3-ketone, Viscum coloratum, Lung neoplasm, A549 cells

中图分类号: 

  • R734.2
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