黄鹌菜的化学成分及其抗肿瘤活性研究  

Chemical composition of Youngia japonica and its antitumor activity

作　　者：莫莉雅 李卫玲[2] 乔嘉璐 彭倩[1] 叶贤胜 孙宾莲 胡康洪[1] MO Liya;LI Weiling;QIAO Jialu;PENG Qian;YE Xiansheng;SUN Binlian;HU Kanghong(Cooperative Innovation Center of Industrial Fermentation,Sino‑German Biomedical Center,Hubei University of Technology,Wuhan 430068,Hubei,China;Wuhan Institute of Biomedical Sciences,School of Medicine,Jianghan University,Wuhan 430056,Hubei,China;Oncology Institute,School of Medicine,Jianghan University,Wuhan 430056,Hubei,China)

机构地区：[1]湖北工业大学中德生物医学中心/工业发酵省部共建协同创新中心,湖北武汉430068 [2]江汉大学医学部武汉生物医学研究院,湖北武汉430056 [3]江汉大学医学部肿瘤研究所,湖北武汉430056

出　　处：《生物资源》2023年第3期267-275,共9页Biotic Resources

基　　金：湖北省重点研发项目(社会发展)(2022BCA018);湖北省自然科学基金项目(2021CFB289);江汉大学科研启动经费(08190001,2021yb138,2019037);江汉大学科研经费(08190006,06210035);工业发酵省部共建协同创新中心开放课题(2022KF16)。

摘　　要：通过硅胶、ODS、Sephadex LH‑20等多种柱色谱技术以及制备HPLC等对黄鹌菜(Youngia japonica)甲醇提取物进行分离,根据核磁波谱技术对分离的化合物进行结构鉴定。利用MTT法筛选影响三阴性乳腺癌(MDA‑MB‑231)细胞生长的化合物,进一步利用克隆形成实验,Hoechst33342/PI双染色法,Annexin V‑FITC/PI双染以及Western blot检测候选化合物对乳腺癌和结直肠癌的抑制效果及可能的作用机制。从黄鹌菜中分离到5个单体化合物,经核磁共振谱鉴定为grosheimin(1),4(‑3'‑hydroxypropyl)‑2,6‑dimethoxyphnol‑3'‑O⁃β⁃D‑glcoside(2),木犀草素‑7‑O‑β⁃D⁃葡萄糖苷(3),木犀草素‑7‑O‑β⁃D⁃葡萄糖醛酸苷(4),芹菜素(5)。通过活性筛选发现grosheimin能显著降低三阴性乳腺癌细胞MDA‑MB‑231的活力。明视野下观察到在不同浓度grosheimin处理下,MDA‑MB‑231和结直肠癌(HCT‑116)细胞形态发生明显改变;grosheimin作用24 h后,MDA‑MB‑231和HCT‑116的IC50分别为16.86μmol/L、30.94μmol/L。克隆形成实验结果显示,在30μmol/L grosheimin的作用下,MDA‑MB‑231和HCT‑116细胞集落形成能力分别为9%、39.7%;Hoechst33342/PI双染法显示,grosheimin可诱导上述两种细胞的凋亡,且存在浓度依赖性。20μmol/L grosheimin作用24 h后,流式细胞仪检测发现MDA‑MB‑231和HCT‑116的凋亡率分别为45%和37.78%;Western blot检测结果显示随着grosheimin处理浓度的增加,凋亡相关蛋白cleaved PARP水平逐渐增加。以上结果表明,grosheimin可能通过诱导细胞凋亡进而抑制MDA‑MB‑231和HCT‑116的生长。The methanol extract of Youngia japonica was separated by column chromatography techniques(silica gel,ODS,Sephadex LH‑20)and preparative HPLC.The structures of the separated compounds were identified using nuclear magnetic spectroscopy.The effect of the compounds on the proliferation of triple‑negative breast cancer(MDAMB‑231)cells was screened by MTT.Furthermore,cloning experiment,Hoechst33342/PI double staining,Annexin VFITC/PI double staining and Western blot were used to detect the anti‑tumor cell effect of the selected compound.A total of five monomeric compounds were isolated from methanol extract of Youngia japonica,the compounds were identified with NMR spectroscopy as grosheimin(1),4(‑3'‑hydroxypropyl)‑2,6‑dimethoxyphnol‑3'‑O⁃β⁃D‑glcoside(2),luteolin‑7‑O⁃β⁃D‑glucoside(3),luteolin‑7‑O⁃β⁃D‑glucuronide(4)and apigenin(5).The anti‑tumor screening assay results showed that the viability of MDA‑MB‑231 cells was significantly reduced by the compound grosheimin.The morphology of MDA‑MB‑231 and colorectal cancer HCT‑116 cells was significantly damaged under different concentrations of grosheimin treatment.The IC50 of MDA‑MB‑231 and HCT‑116 treated by grosheimin were 16.86 and 30.94μmol/L using MTT method.The clone formations of MDA‑MB‑231 and HCT‑116 cells were suppressed to 9%and 39.7%respectively after 30μmol/L grosheimin treatment.Hoechst33342/PI staining results showed that grosheimin could induce the apoptosis of two kinds of cancer cells in a concentration dependent manner,and the apoptosis rates of MDA‑MB‑231 and HCT‑116 were 45%and 37.78%respectively by flow cytometry under 20μmol/L grosheimin treatment of 24 h.Furthermore,the Western blot detection results showed that the level of cleaved PARP was increased also in a grosheimin concentration dependent manner.These results demonstrate that grosheimin may inhibit the growth of triple‑negative breast cancer and colorectal cancer cells via inducing apoptosis.

关 键 词：黄鹌菜 化学成分 grosheimin 抗肿瘤作用 细胞凋亡 

分 类 号：R730.52[医药卫生—肿瘤]