艾叶超临界二氧化碳萃取物的成分鉴定及其抗炎、抗肿瘤活性检测  

ompositional identification of supercritical carbon dioxide extract of Artemisia argyi L.and its anti-inflammatory and anti-tumor activity detection

作　　者：王最 尤月娇 刘闯 张炜[3] 王一飞 王治平 WANG Zui;YOU Yuejiao;LIU Chuang;ZHANG Wei;WANG Yifei;WANG Zhiping(School of Pharmacy∥Guangdong Provincial Key Laboratory for Research and Evaluation of Pharmaceutical Preparations,Guangdong Pharmaceutical University,Guangzhou 510006,Guangdong,China;Tongbai County Tea and Mugwort Industry Development Center,Nanyang 474750,Henan,China;South China Botanical Garden,Chinese Academy of Sciences,Guangzhou 518067,Guangdong,China;Department of Cell Biology,College of Life Science and Technology,Jinan University,Guangzhou 510632,Guangdong,China;Artemisia Argyi Branch Center of National Engineering Research Center for Modernization of Traditional Chinese Medicine,Guangzhou 510632,Guangdong,China;Key Laboratory of Innovative Technology Research on Natural Products and Cosmetics Raw Materials,Guangzhou 510632,Guangdong,China)

机构地区：[1]广东药科大学药学院∥广东省药物新剂型重点实验室,广东广州510006 [2]桐柏县茶艾产业发展中心,河南南阳474750 [3]中国科学院华南植物园,广东广州518067 [4]暨南大学生命科学技术学院细胞生物学系,广东广州510632 [5]国家中药现代化工程技术研究中心艾草分中心,广东广州510632 [6]天然产物化妆品原料创新技术研究重点实验室,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2024年第5期523-538,共16页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金项目(82373917);广州市重点研发计划项目(202206010008);广东药科大学2022年大学生创新训练项目(202210573062)。

摘　　要：目的:针对性分离艾叶(Artemisia argyi L.)超临界二氧化碳萃取物中的萜类化合物,并评估其抗炎、抗肿瘤活性,以期为高效低毒的抗炎、抗肿瘤药物先导物提供基础。方法:将风干的艾叶碾碎后,采用超临界二氧化碳萃取技术提取精油。利用多种分离色谱柱(硅胶柱、ODS、MCI、LH-20、半制备HPLC等)对精油粗提物进行分离纯化,并通过核磁共振波谱技术鉴定分离所得化合物的结构。通过Griess法测定化合物对脂多糖(lipopolysacchride,LPS)诱导的RAW264.7细胞炎症模型的抗炎活性;使用CCK8法测定化合物的抗肿瘤活性及其对正常细胞的毒性。结果:从艾叶中分离得到28个单体化合物,包括3个单萜、21个倍半萜、1个二萜和3个脂肪醇类化合物。化合物4、5、6、9、10、14、15具有抗炎活性,其中化合物4、9与15的抗炎活性显著,其IC 50值分别为(1.8±0.3)μmol/L、(1.2±0.1)μmol/L与(1.5±0.2)μmol/L,CC 50分别是(28.6±1.5)μmol/L、(15.6±0.6)μmol/L和(18.9±0.7)μmol/L。化合物4、5、6、9、14具有抗肿瘤活性,化合物4与5的抗肿瘤活性较为显著,对肿瘤细胞U251与A549的IC 50值分别为(5.9±0.6)μmol/L、(6.3±0.1)μmol/L与(13.5±0.9)μmol/L、(20.5±0.6)μmol/L,对正常细胞BV2的CC 50值分别为(41.0±1.2)μmol/L与(37.4±0.2)μmol/L,具有一定的安全性。结论:从艾叶的超临界二氧化碳萃取物中分离出28个化合物,化合物1、9、11、12、23、24为首次从艾叶中分离得到。化合物4、9与15展现出显著的抗炎活性,而化合物4与5则具有显著的抗肿瘤活性,具有进一步研究的药理价值。Objective:To isolate terpenoids from supercritical carbon dioxide extracts of Artemisia argyi L.and evaluae their anti-inflammatory and anti-tumour activities,in order to provide a basis for the development of highly effective and low-toxicity anti-inflammatory and anti-tumour drug leads.Methods:The essential oil was extracted by supercritical carbon dioxide extraction technique after crushing the air-dried Artemisia argyi leaves.The crude extracts of essential oil were separated and purified using various separation chromatographic columns(silica gel columns,ODS,MCI,LH-20,semi-preparative HPLC,etc).The structures of the isolated compounds were identified by nuclear magnetic resonance spectroscopy.The anti-inflammatory activity of the compounds on the lipopolysaccharide(LPS)-induced RAW264.7 cell inflammation model was determined by the Griess method;the anti-tumor activity of the compounds and their toxicity to normal cells were determined by the CCK8 method.Results:Twenty-eight monomer compounds were isolated from Artemisia argyi leaves,including 3 monoterpenes,21 sesquiterpenes,1 diterpene compound and 3 aliphatic alcohols compounds.Compounds 4,5,6,9,10,14 and 15 were found to have anti-inflammatory activity,among which compounds 4,9 and 15 showed significant anti-inflammatory activity,with IC 50 s of(1.8±0.3)μmol/L,(1.2±0.1)μmol/L,(1.5±0.2)μmol/L,and CC 50 s of(28.6±1.5)μmol/L,(15.6±0.6)μmol/L,(18.9±0.7)μmol/L.Compounds 4,5,6,9 and 14 possessed anti-tumour activity,of which compounds 4 and 5 were more significant,the IC 50 s were(5.9±0.6)μmol/L,(6.3±0.1)μmol/L and(13.5±0.9)μmol/L,(20.5±0.6)μmol/L for tumour cells U251 and A549,and the CC 50 s were(41.0±1.2)μmol/L and(37.4±0.2)μmol/L for normal cells BV2,which provided a certain degree of safety.Conclusion:A total of 28 compounds were isolated from the supercritical carbon dioxide extract of Artemisia argyi L.Compounds 1,9,11,12,23,and 24 were isolated from Artemisia argyi L for the first time.Compounds 4,9,and 15 showed significant ant

关 键 词：艾叶 超临界二氧化碳萃取 萜类化合物 倍半萜内酯 抗炎 抗肿瘤 

分 类 号：R284.2[医药卫生—中药学]