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基于中空纤维细胞垂钓结合高效液相色谱法的茵陈蒿汤主要抗肿瘤成分的体内外研究

Study on major antitumor components in Yinchenhao decoction in vitro and in vivo based on hollow fiber cell fishing coupled with high performance liquid chromatography.

作者信息

Wu Di, Chen Xuan, Hu Shuang, Bai Xiaohong

机构信息

School of Pharmacy, Shanxi Medical University, Taiyuan 030001, China.

School of Pharmacy, Shanxi Medical University, Taiyuan 030001, China.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Aug 15;1060:118-125. doi: 10.1016/j.jchromb.2017.06.003. Epub 2017 Jun 6.

Abstract

Hollow fiber cell fishing (HFCF) based on human hepatoma cell HepG-2 or human renal tubular cell ACHN coupled with high performance liquid chromatography/ultraviolet detection (HPLC/UV) was developed and used for simultaneous study of the major antitumor active components in a formula of Yinchenhao decoction (YCHD) in vitro and in vivo, and in its constituent herbs, namely, Artemisia capillaris Thunb, Gardenia jasminoides Ellis, Radix et Rhizoma Rhei in vitro. Before application, chlorogenic acid, geniposide, p-hydroxyacetophenone, crocin and rhein were chosen as model compounds, the various validations, e.g., cell growth and cell viability on the fiber inner wall, binding between fiber active center and component, repeatability of retention time or relative peak area of the active components were investigated. We screened and identified the major antitumor active components of YCHD, verified their synergetic or antagonistic effect, correlated the major active components between in vitro and in vivo and determined major effective components. Our study will serve as a valuable reference in probing the antitumor material basis of YCHD and its three constituent herbs, as well as in identifying novel clinical aspects of traditional Chinese medicines. The results showed that HCFC-HPLC is a simple, rapid, and reliable method that simultaneously researches the interaction between multiple targets and multiple components of herbal medicine.

摘要

基于人肝癌细胞HepG-2或人肾小管细胞ACHN的中空纤维细胞垂钓(HFCF)结合高效液相色谱/紫外检测(HPLC/UV)技术被开发出来,并用于体外和体内同时研究茵陈蒿汤(YCHD)及其组成草药(即茵陈蒿、栀子、大黄)中的主要抗肿瘤活性成分。在应用之前,选择绿原酸、栀子苷、对羟基苯乙酮、西红花苷和大黄酸作为模型化合物,研究了各种验证指标,如纤维内壁上的细胞生长和细胞活力、纤维活性中心与成分之间的结合、活性成分保留时间或相对峰面积的重复性。我们筛选并鉴定了YCHD的主要抗肿瘤活性成分,验证了它们的协同或拮抗作用,关联了体外和体内的主要活性成分,并确定了主要有效成分。我们的研究将为探究YCHD及其三种组成草药的抗肿瘤物质基础以及确定中药新的临床应用方面提供有价值的参考。结果表明,HFCF-HPLC是一种简单、快速且可靠的方法,可同时研究中药多靶点与多成分之间的相互作用。

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