杨梅叶精油成分分析及其对肺癌A549细胞增殖的抑制作用  

Identification of essential oils in Myrica rubra leaves and their inhibitory effect on the proliferation of lung cancer A549 cell

作　　者：朱丽云[1,2] 张春苗 高永生[1,2] 宋林珍[1] 陈玮 李素芳[1] ZHU Liyun;ZHANG Chunmiao;GAO Yongsheng;SONG Linzhen;CHEN Wei;LI Sufang(Key Laboratory of Marine Food Quality and Hazard Controlling Technology of Zhejiang Province,China Jiliang University,Hangzhou 310018,Zhejiang,China;Anhui Hanfang Biotechnology Co.,Ltd.,Huaibei 235000,Anhui,China;College of Life Sciences,Zhejiang Sci- Tech University,Hangzhou 310018,Zhejiang,China)

机构地区：[1]中国计量大学浙江省海洋食品品质及危害物控制技术重点实验室,杭州310018 [2]安徽汉芳生物科技有限公司,安徽淮北235000 [3]浙江理工大学生命科学学院,杭州310018

出　　处：《果树学报》2018年第7期836-844,共9页Journal of Fruit Science

基　　金：国家自然科学基金面上项目(31571845);浙江省重点科技创新团队(2010R50028)

摘　　要：【目的】比较分析‘丁岙梅’‘东魁’‘晚稻杨梅’和‘荸荠种’4个品种杨梅叶精油的成分与抗癌活性差异。【方法】以水蒸气蒸馏法提取4种杨梅叶精油,采用GC-MS技术分析其组成成分,并以3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐比色法(MTT法)评价了4种杨梅叶精油对肺癌A549细胞增殖的抑制活性。【结果】经GC-MS分析,‘丁岙梅’‘东魁’‘晚稻杨梅’和‘荸荠种’4种杨梅叶精油分别鉴定出34、33、41和33种成分,包括萜烯、萜醇和萜醚3大类主要成分和脂肪醛、烷酮和酚等微量成分。‘丁岙梅’‘东魁’‘晚稻杨梅’和‘荸荠种’4种杨梅叶精油中共有的成分为11种,特有成分分别为7、6、9和5种。β-石竹烯(12.85%~23.87%)、α-葎草烯(19.27%~24.255%)、橙花叔醇(1.15%~23.47%)、氧化香橙烯(3.82%~7.51%)、氧化石竹烯(2.93%~5.26%)和d-杜松烯(0.8%~5.2%)是杨梅叶精油中的主要成分,但含量存在显著差异。根据MTT结果,‘丁岙梅’‘东魁’‘晚稻杨梅’和‘荸荠种’4种杨梅叶精油对肺癌A549细胞增殖抑制作用均存在时间和剂量依赖性,作用24 h后A549细胞半数致死量IC50分别为127.24、127.14、118.03和123.62μg·m L-1,‘晚稻杨梅’叶精油的抗增殖活性略优于其他3种杨梅叶精油。【结论】‘丁岙梅’‘东魁’‘晚稻杨梅’和‘荸荠种’4种杨梅叶精油成分差异显著,对肺癌A546细胞增殖抑制活性较强,不同品种间无显著差异(p>0.05)。【Objective】The secondary metabolites of essential oils in Myrica rubra leaves（MEO） have antioxidant, antibacterial, anticancer and other biological activities. In this paper, the chemical compositions and anti-cancer activity of MEO extracted from the leaves of four bayberry cultivars,‘Ding＇ao＇‘Dongkui＇‘Wandao＇and‘Biqi＇, were analyzed. This study could provide a theoretical basis for the development of natural anti-cancer drugs.【Methods】The essential oils were extracted with steam distillation from the leaves of the four cultivars,‘Ding＇ao＇（grown in Wenzhou, Zhejiang）,‘Dongkui＇（grown in Xianju, Zhejiang）,‘Wandao＇（grown in Zhoushan, Zhejiang） and‘Biqi＇（grown in Yuyao,Zhejiang）. GC-MS was used to analyze the components and contents of essential oils. The chromatographic analysis column was a HP-5 type（30 m×0.25 mm×0.25 μ m）. The initial was 50 ℃, which was kept for 1 min, rose to 250 degrees at 8 min and then maintained for 5 min. Carrier gas was high purity helium with a flow rate of 1 m L· min-1. The temperature of the vaporizing chamber was 220 ℃. The in-jection volume was 1 μL. The split ratio was 20：1. Mass spectrometry conditions included an EI source with an ion source temperature of 200 ℃, a solvent delay time of 2 min, and a mass range of 50-500 u.Components were indentified based on the NIST＇08 standard mass spectrometry database and related literatures. The relative GC contents of volatile oils were calculated by area normalization method. And then the compositions and contents of essential oils were comparatively analyzed. Human lung cancer A549 cells were used as the material to examine the inhibitory activities of four MEOs on the proliferation of the lung cancer A549 cells using 3-（4, 5-Dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide assay（MTT assay）. The concentrations of MEOs used were 50, 75, 100, 125, 150, 175 and 200 μ g· m L-1.The action durations were 24, 48, 72, 96 and 120 h, and the proliferat

关 键 词：杨梅叶精油 成分分析 3-(4 5-二甲基噻唑-2)-2 5-二苯基四氮唑溴盐MTT 肺癌细胞 增殖抑制 

分 类 号：S667.6[农业科学—果树学]