摘要
目的筛选口虾蛄总生物碱的提取方法,并研究口虾蛄总生物碱在体外对人鼻咽癌CNE-2Z细胞的抑制作用。方法口虾蛄乙醇提取物及新鲜口虾蛄分别以酸提碱沉,继以乙酸乙酯萃取,取得上清提取物(简称L)和沉淀提取物(简称S);以薄层色谱法探讨各提取物的分离条件及初步化学性质鉴定;继用CCK-8法观察各提取物体外对人鼻咽癌CNE-2Z细胞生长增殖的影响。结果以新鲜口虾蛄酸提碱沉后萃取所得总生物碱较多;氯仿-环乙烷系统分离各提取物效果最佳;沉淀提取物(S)的显色反应及色谱显色结果显示其可能为生物碱类物质;口虾蛄总生物碱S1和S2均对人鼻咽癌CNE-2Z细胞有明显的抑制作用,而上清提取物L1及L2则不明显;口虾蛄总生物碱S1和S2作用于人鼻咽癌CNE-2Z细胞48 h后,其IC_(50)分别为(71.18±0.56)μg/mL和(49.31±0.65)μg/mL。结论以新鲜口虾蛄酸提碱沉后萃取总生物碱的方法,简便、稳定、高效;口虾蛄总生物碱体外可明显抑制人鼻咽癌CNE-2Z细胞生长,显示有较好的开发应用前景。
Objective To screen the extraction method to obtain the total alkaloids of Squilla oratoria, and investigate the effects of the total alkaloids on nasopharyngeal carcinoma cell line CNE-2Z. Methods The acid extraction and alkaline deposition, and ethyl acetate extraction were used in the extraction and separation of the ethanol extract of Squilla oratoria and fresh Squilla oratoria, and then the supernatant extract （L for short） and deposition extract （S for short） were collected. Thin- layer chromatography （TLC） and chemical reactions were used to explore the deploying condition and determine the nature preliminarily. Following, the CCK-8 method was used to observe each extract on the growth and proliferation of human nasopharyngeal carcinoma CNE-2Z ceils in vitro. Results The maximum alkaloids were extracted with the acid extraction and alkaline deposition, and ethyl acetate extraction was directly extracted from fresh Squilla oratoria. The chloroform- cyclohexane system was the best deploying system for the total alkaloids of Squilla oratoria. The results of color identification by TLC showed that the precipitation extract （S for short） belong to alkaloids. Through the CCK-8 method screening, it was discovered that S1 and $2 had the strong inhibitory action on the human nasopharyngeal carcinoma CNE-2Z cell growth, but the function of L1 and L2 relatives was not obvious. The IC50（s） of cell viability on nasopharyngeal carcinoma CNE-2Z cells after treated with the total alkaloids of Squilla oratoria S 1 and S2 for 48 h were （71.18±0.56） μg/mL and （49.31 ± 0.65）μg/mL, respectively. Conclusions The method of extraction of the total alkaloids from fresh Squilla oratoria by the acid extraction and alkaline deposition, and ethyl acetate extraction is stable, convenient and efficient. The total alkaloids of Squilla oratoria have an inhibitory effect on the cell growth of CNE-2Z cells, which has a good development and application prospect.
作者
王槐高
孔霞
顾帝水
郭向华
周艳芳
WANG Huaigao KONG Xia GU Dishui GUO Xianghua ZHOU Yanfang(Department of Pathophysiology, Guangdong Medical University, Dongguan, Guangdong 523808, Chin)
出处
《中国热带医学》
CAS
2017年第5期449-451,共3页
China Tropical Medicine
基金
广东建设中医药强省科研项目(No.20141155)
广东医学院面上基金项目(No.2xk1310)
关键词
口虾蛄
总生物碱
酸提碱沉法
抗肿瘤活性
Squilla oratoria
alkaloids
extraction and alkaline deposition
antitumor activity
