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

doi:10.6040/j.issn.1671-7554.0.2017.070

摘要/Abstract

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

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R734.2

引用本文

黄竹青,吴雪韦,任冬梅. 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[J]. 山东大学学报（医学版）, 2017, 55(8): 35-41.

HUANG Zhuqing, WU Xuewei, REN Dongmei. Isolation and identification of phenolic constituents from Viscum coloratum and the antiproliferative effects on A549 cells[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2017, 55(8): 35-41.

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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

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

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[1]	于嘉智王克来庄岩张连海. S-100和C-kit蛋白在儿童先天性巨输尿管中的表达[J]. 山东大学学报(医学版), 2209, 47(6): 73-.
[2]	张莹王宏伟王慧陈福琴. Graves病复发患者外周血中转化生长因子β1水平的变化[J]. 山东大学学报(医学版), 2209, 47(6): 80-.
[3]	栗英林,宋道庆,徐忠华. 应用生物信息学方法分析肾透明细胞癌中FKBP11的表达[J]. 山东大学学报 (医学版), 2020, 1(9): 45-51.
[4]	孟朝暾,李钦, 孙卉,邬信芳,孙文凯,高伟. 白细胞介素18单核苷酸多态性与73例喉鳞状细胞癌易感性的关联性[J]. 山东大学学报 (医学版), 2020, 1(9): 58-63.
[5]	宋立,张艳,刘洋,王丹. 随访2年观察1例新发突变的营养不良型大疱性表皮松解症[J]. 山东大学学报 (医学版), 2020, 1(8): 120-122.
[6]	张宝文,雷香丽,李瑾娜,罗湘俊,邹容. miR-21-5p靶向调控TIMP3抑制2型糖尿病肾病小鼠肾脏系膜细胞增殖及细胞外基质堆积[J]. 山东大学学报 (医学版), 2020, 1(7): 7-14.
[7]	马青源,蒲沛东,韩飞,王超,朱洲均,王维山,史晨辉. miR-27b-3p调控SMAD1对骨肉瘤细胞增殖、迁移和侵袭作用的影响[J]. 山东大学学报 (医学版), 2020, 1(7): 32-37.
[8]	肖娟,肖强,丛伟,李婷,丁守銮,张媛,邵纯纯,吴梅,刘佳宁,贾红英. 两种甲状腺超声数据报告系统诊断效能的比较[J]. 山东大学学报 (医学版), 2020, 1(7): 53-59.
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[10]	赵姗姗,李晓梅,梁婷,张超,宋静,侯桂华. 自噬对¹²⁵I-anti-TLR5同种移植排斥靶向显像的影响[J]. 山东大学学报（医学版）, 2017, 55(9): 46-52.
[11]	林雪艳,李春艳,侯小满,田永杰. PARP-1及EMT标志物在子宫腺肌病在位及异位内膜中的表达[J]. 山东大学学报（医学版）, 2017, 55(9): 36-40.
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[14]	胡玉敬,吴大勇,张文艳,边艳珠,魏强,田丛娜,常胜利. 放疗后肿瘤⁹⁹Tc^m-MNLS乏氧显像变化与乏氧诱导因子-1α表达的相关性[J]. 山东大学学报（医学版）, 2017, 55(8): 30-34.
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