柴胡桂枝汤挥发油的GC-MS分析及对人肺腺癌A549细胞体外增殖的抑制作用研究  

GC-MS Analysis of Volatile Oil from Chaihu Guizhi Decoction and Study on Inhibitory Effects on in vitro Proliferation of Human Lung Adenocarcinoma A549 Cells

作　　者：文冉 李壮壮 杜以晴 杨勇[1] 容蓉[1] 吕青涛[1] WEN Ran;LI Zhuangzhuang;DU Yiqing;YANG Yong;RONG Rong;LYU Qingtao(College of Pharmacy,Shandong University of TCM,Jinan 250355,China;College of Pharmacy,Ocean University of China,Shandong Qingdao 266100,China)

机构地区：[1]山东中医药大学药学院,济南250355 [2]中国海洋大学药学院,山东青岛266100

出　　处：《中国药房》2021年第1期29-33,共5页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81873220,No.81774167);山东省重大科技创新工程项目(No.2018CXGC1307)。

摘　　要：目的:研究复方柴胡桂枝汤挥发油的组成成分,并评价其对人肺腺癌A549细胞体外增殖的抑制作用。方法:根据2015年版《中国药典》(四部)通则2204挥发油测定法中甲法提取柴胡桂枝汤中的挥发油。采用气相色谱-质谱联用法(GC-MS)结合Kováts指数分析其挥发油成分,并采用峰面积归一化法计算各成分的相对含量。以不同质量浓度顺铂(4、8、16、32、64 mg/L)为阳性对照,采用MTT法检测不同质量浓度柴胡桂枝汤挥发油(25、50、100、200、400 mg/L)作用48 h后对A549细胞体外增殖的抑制作用;另设阴性对照组(加细胞但不加药物)。结果:从挥发油中共分离出71个成分,鉴定出其中59个成分,其峰面积之和占总峰面积的84.99%。其中,相对含量较高的成分为芳姜黄烯(17.65%)、β-甜没药烯(9.57%)、β-罗勒烯(7.05%)、α-姜黄烯(5.35%)、2,5-二甲基苯甲醛(4.24%)、异丁酸芳樟酯(2.70%)、α-雪松烯(2.48%)、δ-杜松烯(2.07%)。与阴性对照组比较,4~64 mg/L顺铂组和25~400 mg/L柴胡桂枝汤挥发油组细胞的增殖率均显著降低(P<0.05);顺铂和柴胡桂枝汤挥发油对A549细胞体外增殖的半数抑制浓度分别为10.150、73.526 mg/L。结论:柴胡桂枝汤挥发油中主要以芳姜黄烯、β-甜没药烯、β-罗勒烯、α-姜黄烯等成分为主,其对A549细胞的体外增殖具有一定的抑制作用。OBJECTIVE:To study the composition of the volatile oil from Compound chaihu guizhi decoction,and to evaluate its in vitro anti-proliferative activity on human lung adenocarcinoma A549 cells.METHODS:The volatile oil from Chaihu guizhi decoction was extracted according to the steam distillation method of general rules 2004 in the 2015 edition of Chinese Pharmacopoeia(partⅣ).The volatile oil components were analyzed by GC-MS combined with Kováts index,and the relative content of each component was calculated by peak area normalization method.Using different concentrations of cisplatin(4,8,16,32,64 mg/L)as positive control,MTT assay was used to detect the inhibitory effects of different concentrations of volatile oil from Chaihu guizhi decoction(25,50,100,200,400 mg/L)on in vitro proliferation of A549 cell after 48 h of treatment.Negative control group(with cells but without drugs)was set up.RESULTS:A total of 71 chemical components were isolated from the volatile oil,among which there were 59 compounds identified,sum of peak areas accounting for 84.99%of the total peak area.The compounds with relatively high content included ar-curcumene(17.65%),β-bisabolene(9.57%),β-ocimene(7.05%),α-curcumene(5.35%),2,5-dimethylbenzaldehyde(4.24%),linalyl isobutyrate(2.70%),α-cedrene(2.48%),δ-cadinene(2.07%).Compared with negative control group,the proliferation rate of cells were decreased significantly in 4-64 mg/L cisplatin groups and 25-400 mg/L volatile oil from Chaihu guizhi decoction groups(P<0.05).IC50 of cisplatin and volatile oil from Chaihu guizhi decoction to in vitro proliferation of A549 cells were 10.150 and 73.526 mg/L.CONCLUSIONS:The volatile oil from Chaihu guizhi decoction mainly includes ar-curcumene,β-bisabolene,β-ocimene,α-curcumene,which shows certain inhibitory effect on in vitro proliferation of A549 cells.

关 键 词：柴胡桂枝汤 挥发油 气相色谱-质谱联用法 Kováts指数 人肺腺癌A549细胞 

分 类 号：R285[医药卫生—中药学] R932[医药卫生—中医学]