鹅不食草中抗肺癌活性成分筛选及异戊酸心菊内酯单体的体外作用研究  

Screening of anti-lung cancer compounds and in vitro mechanism of helenalin-isovalerate for Centipeda minima

作　　者：胡倩 薛鹏辉 于弘 王敏 赵新春 邱峰[4] 周光飚 HU Qian;XUEPeng-hui;YU Hong;WANG Min;ZHAO Xin-chun;QIU Feng;ZHOU Guang-biao(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;State Key Laboratory of Molecular Oncology,National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100021,China;State Key Laboratory of Membrane Biology,Institute of Zoology,Chinese Academy of Sciences,Beijing 100101,China;School of Chinese Materia Medica,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]国家癌症中心/中国医学科学院北京协和医学院肿瘤医院分子肿瘤学国家重点实验室,北京100021 [3]中国科学院动物研究所膜生物学国家重点实验室,北京100101 [4]天津中医药大学中药学院,天津301617

出　　处：《中成药》2020年第12期3151-3157,共7页Chinese Traditional Patent Medicine

基　　金：国家杰出青年基金(81425025)。

摘　　要：目的在32个鹅不食草单体化合物中筛选具有抗肺癌活性的有效成分,并探究其单体异戊酸心菊内酯的体外抗肺癌作用。方法 MTT比色法检测化合物对5种人肺癌细胞系的增殖抑制作用,筛选出活性较好的化合物,流式细胞术检测细胞凋亡,Western blot检测p-ERK、ERK、p-AKT、AKT、NF-κB表达。结果有10种(31.3%)成分在20μmol/L浓度下能使H460、A549细胞的增殖活性抑制15%以上。阿里二醇、6-氧-当归酰多梗白菜菊素、异戊酸心菊内酯在2.5~20μmol/L浓度范围内对肺癌细胞具有较明显的抑制作用,随着化合物浓度的升高,细胞增殖抑制作用增强,诱导肺癌细胞凋亡的作用也增强(P<0.01)。Western blot结果显示,与对照组比较异戊酸心菊内酯能以浓度依赖性的方式活化caspase-3、caspase-8,使其活化片段表达增高(P<0.01),而caspase-3底物PARP被降解,同时显著抑制了H1975细胞增殖相关蛋白p-ERK、p-AKT及核转录因子NF-κB的表达(P<0.05)。结论鹅不食草含有多种具有抗肺癌活性的成分,能显著抑制肺癌细胞增殖并诱导其凋亡,其作用机制可能与抑制p-ERK、p-AKT及NF-κB的表达有关。AIM Based on our previous study of 6-O-angeloylplenolin(6-OAP),a component of the medicinal herb Centipeda minima(L.)A.Br.et Aschers.,with strong anti-lung cancer activity in vitro and in vivo,here we tried to investigate the effects of 32 compounds isolated from Centipeda minima toward five lung cancer cell lines to identify their anti-lung activity and to explore the anti-lung cancer mechanism of helenalin-isovalerate in vitro.METHODS The effects of compounds on cell proliferation of H460,A549,H1975,H1299,and H520 cells were tested by 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-diphenytetrazoliumromide(MTT)assays for the IC50 values determination.The apoptotic cells were analyzed by Annexin V-propidium iodide(PI)method,and changes in PARP,caspase-3,caspase-8,p-ERK,p-AKT and NF-κB were evaluated by Western blot.RESULTS Among the 32 compounds,10 ones(31.3%)were able to suppress cell proliferation by 15%or more.Three compounds,arnidiol,6-O-angeloylplenolin and helenalin-isovalerate,showed significant inhibition on the five lines with relatively low IC50 values.We observed that helenalin-isovalerate not only induced apoptosis of H1975 and H1299 cells with cleavage of PARP and activation of Caspase-3 and Caspase-8(P<0.01),but also inhibited the expression of p-AKT,p-ERK and NF-κB in H1975 cells(P<0.05).CONCLUSION Centipeda minima bears several components that are significantly inhibitory toward lung cancer cells through inhibition of cell proliferation and induction of cell apoptosis.The mechanism of this medicinal herb in treating lung cancer may depend on inhibiting the expressions of p-AKT,p-ERK and NF-κB.

关 键 词：鹅不食草 异戊酸心菊内酯 肺癌 增殖抑制 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]