金银花的多酚粗提物诱导人肺癌H1299细胞凋亡作用及机制研究  

The effect and mechanism of polyphenolic extract from Lonicera japonica on apoptosis promotion of lung cancer cells in vitro

作　　者：陈垒 张伟 杜亚明 Chen Lei;Zhang Wei;Du Yarning(The first affiliated hospital of Jinzhou medical university,Jinzhou 121000)

机构地区：[1]锦州医科大学附属第一医院,锦州121000

出　　处：《中药药理与临床》2018年第3期89-93,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：辽宁省科学技术厅重点研发计划指导计划项目(项目编号;2017225044);辽宁省省直医院改革重点临床科室诊疗能力建设项目青年项目任务书(项目编号;LNCCC-D29-2015)

摘　　要：目的:通过观察金银花多酚粗提物对肺癌细胞增殖和凋亡的影响,探究其诱导凋亡可能的分子机制。方法:HPLCMS分析金银花多酚粗提物主要化学成分;以人肺癌细胞H1299细胞为研究对象,使用不同浓度(100、400、800、1200、1500μg/ml)的金银花多酚粗提物作用于肺癌H1299细胞;CCK-8法检测细胞存活率情况;流式细胞术分析细胞凋亡水平和细胞周期分布;Western blot检测Bcl-2、Bax、Procaspase-3、Procaspase-6、Procaspase-8、Procaspase-9和PARP蛋白的表达。结果:金银花多酚粗提物主要包含18种多酚类化合物。与空白对照组相比,金银花多酚浓度高于100μg/ml时,H1299细胞增殖受到明显抑制;金银花多酚浓度高于400μg/ml时,H1299细胞凋亡水平明显升高;1200μg/ml金银花多酚作用时,sub G1期细胞的数量明显增高,而G1期、S期、G2/M期细胞数目均明显下降;金银花多酚浓度高于1200μg/ml时,Bcl-2表达水平明显下降,Bax表达表达水平明显上升;金银花浓度高于400μg/ml时,Procaspase-3、Procaspase-6、Procaspase-8和Procaspase-9表达水平均明显下降。结论:金银花多酚粗提物能够抑制肺癌细胞增殖,促进肺癌细胞凋亡,诱导细胞周期阻滞,其机制可能与下调Bcl-2的蛋白表达,上调Bax的蛋白表达,激活caspase级联反应有关。Objective： To investigate the effect and mechanism of polyphenolic extract from Lonicera japonica on apoptosis promotion of lung cancer cells in vitro. Methods： The lung cancer cell line H1299 cells were treated with different concentrations（100,400,800,1200,1500 μg/ml）of polyphenolic extract from Lonicera japonica. HPLC-MS was used to analyze the main chemical constituents of polyphenolic extract,CCK-8 method was used to detect the viability of lung cancer cells,flow cytometry was applied to detect apoptosis and cell cycle distribution of lung cancer cells,western blotting method was used to detect protein expressions of Bcl-2,Bax,Procaspase-3,Procaspase-6,Procaspase-8 and Procaspase-9 in lung cancer cells. Results： Polyphenolic extract from Lonicera japonica mainly contains 18 polyphenols. Compared with the control group,the viability of H1299 lung cancer cells was significantly decreased after the treatment with polyphenolic extract over 100 μg/ml（P〈0. 05）. Apoptosis rates of H1299 lung cancer cells were significantly increased after the treatment with polyphenolic extract over 400μg/ml（P〈0. 05）,the distribution of sub-G1 phase cells was significantly increased after the treatment with 1200 μg/ml polyphenolic extract（P〈0. 05）,the protein expression of Bcl-2 was significantly decreased after the treatment with 1200 μg/ml polyphenolic extract（P〈0. 05）,protein expression levels of Procaspase-3,Procaspase-6,Procaspase-8 and Procaspase-9 were significantly decreased after the treatment with polyphenolic extract over 400 μg/ml（P〈0. 05）. Conclusion： Polyphenolic extract from Lonicera japonica has the significant inhibitory effect on the viability of lung cancer cells,including inducing cell apoptosis. Its possible underlying mechanism may be related to regulating protein expressions of Bcl-2,Bax,and activate caspase pathway.

关 键 词：金银花 多酚 肺癌 凋亡 凋亡蛋白酶 

分 类 号：R285[医药卫生—中药学]