蓝萼香茶菜中具有吲哚胺2,3-双加氧酶(IDO)抑制活性成分的研究  

Isolation and purification of indoleamine 2,3-dioxygenase(IDO) inhibitory constituents from Rabdosia japonica

作　　者：王辉俊 朱加明[4] 韩俊琦 田浩威 陈炜 叶冠[1] WANG Hui-jun;ZHU Jia-ming;HAN Jun-qi;TIAN Hao-wei;CHEN Wei;YE Guan(Central Research Institute,Shanghai Pharmaceuticals Group Holding Co.,Ltd.,Shanghai 201203,China;Key Laboratory for Standardization of Chinese Medicines of Ministry of Education,Institute of Chinese Materia Medica,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai R&D Center for Standardization of Chinese Medicines,Shanghai 201203,China;Hangzhou Huqingyutang Pharmaceutical Co.,Ltd.,Hangzhou 311199,China)

机构地区：[1]上海医药集团股份有限公司中央研究院,上海201203 [2]上海中医药大学中药研究所中药标准化教育部重点实验室,上海201203 [3]上海中药标准化研究中心,上海201203 [4]杭州胡庆余堂药业有限公司,浙江杭州311199

出　　处：《中草药》2020年第17期4419-4424,共6页Chinese Traditional and Herbal Drugs

基　　金：上海市科委科研计划项目(15DZ1900100);上海市科委科研计划项目(15DZ1900104);上海市自然科学基金(20ZR1458100);上海市青年科技启明星计划资助(19QB1406500)。

摘　　要：目的基于活性导向分离蓝萼香茶菜Rabdosia japonica中具有吲哚胺2,3-双加氧酶(IDO)抑制作用的成分。方法蓝萼香茶菜地上部分经水提醇沉后得粗多糖XPS,其经活性跟踪测定、DEAESepharoseFastFlow阴离子交换树脂分离和Superdex-75系列凝胶纯化后得XPS10-1,并对XPS10-1进行HPGPC法分析、单糖组成测定、氨基酸组成分析和IDO抑制活性检测。结果从蓝萼香茶菜中得到一种具有IDO抑制作用的均一糖蛋白成分XPS10-1,相对分子质量为8 852,糖基部分主要由鼠李糖基和葡萄糖基组成,物质的量比为10.0∶2.2,蛋白部分主要由谷氨酸、丝氨酸和甘氨酸组成,物质的量比为37.3∶16.9∶45.8,XPS10-1表现出较强的IDO抑制活性,半数抑制浓度(IC50)值为(46.6±3.4)μg/mL,XPS10-1对人宫颈癌He La细胞中IDO酶也有抑制作用,IC50值为(139.0±8.7)μg/mL。结论从蓝萼香茶菜中成功分离到具有IDO抑制作用的糖蛋白,为蓝萼香茶菜的化学物质基础提供依据。Objective To isolate indoleamine 2,3-dioxygenase(IDO) inhibitory constituents from Rabdosia japonica based on bioactivity tracking separation. Methods The overground part of R. japonica was extracted with boiling water and precipitated by ethanol, the precipitation was collected and lyophilized to obtain XPS, then successively separated by DEAE Sepharose Fast Flow anion-exchange and Superdex-75 gel permeation chromatographic steps to give XPS10-1. A combination of HPGPC, monosaccharide and amino acid composition analysis and IDO inhibitory studies was performed to investigate the structure and bioactivity of XPS10-1. Results A IDO inhibitory glycoprotein, XPS10-1, was obtained from R. japonica based on activity tracking, its average molecular weight was estimated to 8 852, monosaccharide composition analysis showed the glycosyl part of XPS10-1 was mainly composed of rhamnose and glucose with the ratio of 10.0:2.2, and the protein part was mainly composed of glutamic acid, serine and glycine with mass ratio of 37.3:16.9:45.8. XPS10-1 showed potent IDO inhibitory effect with IC50 of(46.6 ± 3.4) μg/mL, and IC50 of IDO inhibitory effect of XPS10-1 on HeLa cells was(139.0 ± 8.7) μg/mL. Conclusion In this study, a glycoprotein with IDO inhibitory effect was isolated from R. japonica, which could lay the foundation for the substance basis study of R. japonica.

关 键 词：蓝萼香茶菜 糖蛋白 活性导向分离 吲哚胺2 3-双加氧酶(IDO) 抑制作用 

分 类 号：R284.1[医药卫生—中药学]