基于线粒体的离心超滤/液相色谱/质谱法筛选中药线粒体靶向活性成分
Screening Mitochondria-targeting Compounds from Traditional Chinese MedicinesUsing a Mitochondria-based Centrifugal Ultrafiltration/Liquid Chromatography/Mass Spectrometry Method
格根塔娜 王璇 尚明英 杨兴鑫 徐风 俞捷 周瑜珍 蔡少青
作者信息
摘要
线粒体因在能量生成、细胞凋亡、氧化应激、钙稳态和脂质代谢中均发挥核心作用，其已成为多种疾病治疗的重要靶点而日渐备受关注。为此，我们利用一种基于线粒体的离心超滤与LC/MS联用的方法有效筛选中药（葛根和川芎）中的线粒体靶向化合物，这种方法被称之为线粒体靶向活性成分筛选方法（SM-MBC），适用于复杂基质中线粒体靶向化合物的筛选。采用此法从葛根和川芎2种中药提取物中成功筛选到23种活性化合物，采用LC/MS鉴定了17种，其中有12种是新发现的线粒体靶向化合物。采用离体药理实验发现有7种被鉴定的化合物具有调节线粒体功能的作用。此外，还采用细胞实验首次揭示了部分被筛选化合物对HepG2细胞增殖和心肌细胞活力降低（缺氧/复氧所致）的抑制作用。研究结果将有助于深入阐释中药治疗线粒体相关疾病的机理，以及利用中药来源的先导化合物研发药物；且结果也进一步证明SM-MBC可作为一种从复杂样品（如葛根和川芎提取物）中快速筛选线粒体靶向化合物的有效方法。
Abstract
Mitochondria are becoming of increasing interest as a highly significant target for various disease remedy over the years, because mitochondria play a central role in energy production, apoptotic cell death, oxidative stress, calcium homeostasis and lipidic metabolism. We thus applied an efficient mitochondria-based centrifugal ultrafiltration/liquid chromatography/mass spectrometry method, which is called screening method for mitochondria-targeted bioactive constituents (SM-MBC) and is agree with the search of mitochondria-targeting compounds from complex matrix, to hunt for natural mitochondria-targeting compounds from traditional Chinese medicines (TCMs) including Puerariae Radix (PR) and Chuanxiong Radix (CR). Totally 23 active compouds were successfully discovered from the two extracts of TCMs. Among the 23 hit compounds, 17 were identified by LC/MS, of which 12 were new mitochondria-targeting compounds. The abilities of 7 of the 17 identified active compounds to remedy mitochondrial functions were exposed by in vitro pharmacological tests. Furthermore, the inhibition capabilities of the hit compounds on HepG2 cell proliferation and on loss of cardiomyocyte viability induced by hypoxia/reoxygenation injury were first exposed by cell tests. The results obtained are benefit for deep-going elucidation of the therapeutic principles of TCMs used to remedy mitochondria-involved disorders and drug development using TCMs as lead compounds, and SM-MBC was further showed to be an efficient protocol for the rapid screening of mitochondria-targeting constituents from complex preparation such as PR and CR extracts.关键词
线粒体/活性成分/中药/离心超滤/液相色谱-质谱联用Key words
Mitochondria/Bioactive constituents/Traditional Chinese medicines/Centrifugal ultrafiltration/Liquid chromatography/mass spectrometry引用本文复制引用
格根塔娜,王璇,尚明英,杨兴鑫,徐风,俞捷,周瑜珍,蔡少青.基于线粒体的离心超滤/液相色谱/质谱法筛选中药线粒体靶向活性成分[EB/OL].(2017-07-10)[2026-04-05].http://www.paper.edu.cn/releasepaper/content/201707-29.学科分类
药学/基础医学/中医学
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