臭牡丹地上部分抗肿瘤活性的筛选  

Screening for compounds with antitumor activity in the aerial part of Clerodendrum bungei

作　　者：余娜[1] 朱克俭[2] 谢谊[2] 

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南省中医药研究院,湖南长沙410006

出　　处：《湖南中医杂志》2017年第11期147-150,共4页Hunan Journal of Traditional Chinese Medicine

基　　金：湖南省自然科学基金资助项目(编号:14JJ4066);湖南省教育厅资助项目(编号:13C674)

摘　　要：目的:对臭牡丹地上部分所含的化合物进行抗肿瘤活性的筛选。方法:采用70%乙醇加热回流提取臭牡丹的地上部分,通过石油醚、二氯甲烷、正丁醇依次萃取和大孔吸附树脂、硅胶柱色谱进行分离纯化;采用液质联用技术对臭牡丹地上部分的粗分化合物进行了分析;采用MTT法对臭牡丹地上部分的不同萃取物质进行体外抗肿瘤活性的筛选;采用CCK8法检测毛蕊花糖苷和粗分物对A549细胞增殖的影响。结果:通过液质分析结果和文献比较,确定粗分物中是以毛蕊花糖苷为主的苯乙醇苷类化合物;95%乙醇洗脱部分的IC50为0.019mg/m L,二氯甲烷部分、25%乙醇及50%乙醇的IC50分别为0.039、0.180和0.181mg/m L,水部分的IC50不存在;毛蕊花糖苷的CC50值约为0.22mg/m L,粗分物A的CC50值约为1.3mg/m L。结论:MTT结果显示,95%乙醇部分抗肿瘤活性最好,其次为二氯甲烷,25%乙醇与50%乙醇第三,水部分基本没作用;CCK8结果显示,毛蕊花糖苷对A549细胞增殖有明显抑制作用,且具有浓度依赖性;粗分物A对A549细胞有低浓度促进、高浓度抑制的作用。Objective ：To screen out the compounds with antitumor activity in the aerial part of Clerodendrum bungei. Methods：Heating reflux extraction with 70% alcohol was performed for the aerial part of Clerodendrum bungei, and isolation and purification was performed by extraction with petroleum ether, dichloromethane, and n - butyl alcohol in sequence, macroporous adsorption resin, and silica gel column chromatography. Liquid chromatogra- phy- mass spectrometry was performed to analyze the crude compounds in the aerial part of Clerodendrum bungei. MTT was used to screen out the compounds which were extracted from the aerial part of Clerodendrum bungei and had an in vitro antitumor activity. CCK8 assay was used to investigate the effect of verbascoside and crude compounds on the proliferation of A549 cells. Results：With reference to the results of liquid chromatography -mass spectrometry and related literature, the crude compounds were phenylethanoid glycosides, mainly verbascoside. The eluted fraction of 95% alcohol had a half-maximal inhibitory concentration（IC50）of 0. 019 mg/ml;the eluted fractions of dichloromethane,25 % alcohol, and 50% alcohol had an IC50 of 0. 039,0. 180, and 0. 181 mg/ ml,respectively;the water -eluted fraction did not have an IC50. Verbascoside had a 50% cytotoxic concentration （CC50） of 0.22 mg/ml, and crude compound A had a CC50 of 1.3mg/mL Conclusion：According to the results of MTT, the eluted fraction of 95 % alcohol has the best antitumor activity, followed by the eluted fractions of dichlo- romethane ,25% alcohol,and 50% alcohol, and the water- eluted fraction almost has no antitumor activity. Based on the results of CCK8, verbascoside has a significant inhibitory effect on the proliferation of A549 cells in a concentration -dependent manner. Crude compound A promotes the proliferation of A549 ceils at a low concentration and inhibits the proliferation at a high concentration.

关 键 词：臭牡丹 抗肿瘤 液质联用 MTT CCK8 实验研究 

分 类 号：R285.5[医药卫生—中药学]