State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao SAR, 999078, China.
School of Chemistry and Chemical Engineering, Chongqing University, Chongqing, 401331, China.
J Ethnopharmacol. 2022 Jun 28;292:115197. doi: 10.1016/j.jep.2022.115197. Epub 2022 Mar 21.
Salvia miltiorrhiza (Danshen, DS), the dried root and rhizome of Salvia miltiorrhiza Bunge and Ligusticum chuanxiong (Chuanxiong, CX), the dried rhizomes of Ligusticum striatum DC are effective in invigorating blood circulation and eliminating stasis which is highly related with cardiovascular disease (CVD).
The identification of activity-based chemical markers is very important, but the complex mechanism of "multi-component, multi-target, and multi-effect" within traditional Chinese medicine (TCM) poses a great challenge to this work. In this study, we combined network pharmacological prediction with experimental validation of the DS and CX to explore an effective method for discovering quality control (QC) of antithrombotic herbs by clarifying the intermediate layer "module/cluster" between the whole complex system and a single component.
Based on structural similarity analysis of compound and the thrombosis network published before, we firstly modularized two layers called chemical cluster (CC) network and functional module (FM) network respectively and linked them into one bilayer modularized compound target (BMCT) network. "Two-step" calculation was applied on identifying the significant compounds as the potential QC markers from CC. The in vitro inhibitory activity of selected QC marker compounds on thrombin was evaluated to partially verify their pharmacological activities. HPLC was used to determine contents.
According to the network-based analysis, nine compounds with great importance in the BMCT network were identified as QC markers of DS-CX, including tanshinone I, tanshinone IIA, cryptotanshinone, salvianolic acid B, ferulic acid, salvianolic acid A, rosmarinic acid, chlorogenic acid, and coniferyl ferulate. Enzyme inhibitory test partially verified the activity of tanshinone I and tanshinone IIA. Chemical profiling indicated that the nine marker compounds are the main components in the herbal pair.
This study identified activity-based QC markers of DS-CX herbal pair and provided a new methodology that can be used in the QC of other herbs, herbal pairs, or formulas.
丹参（丹参，DS），丹参的干燥根和根茎，川芎（川芎，CX），藁本的干燥根茎，是一种有效的活血化瘀药，与心血管疾病（CVD）高度相关。
基于活性的化学标志物的鉴定非常重要，但中药（TCM）中“多成分、多靶点、多效”的复杂机制对此项工作提出了巨大挑战。在这项研究中，我们结合网络药理学预测与 DS 和 CX 的实验验证，通过阐明整个复杂系统与单个成分之间的中间层“模块/群集”，探索发现抗血栓草药质量控制（QC）的有效方法。
基于化合物的结构相似性分析和之前发表的血栓形成网络，我们首先将两个层分别模块化，称为化学簇（CC）网络和功能模块（FM）网络，并将它们链接成一个双层模块化化合物靶（BMCT）网络。“两步”计算用于从 CC 中识别出作为潜在 QC 标志物的显著化合物。选择的 QC 标志物化合物对凝血酶的体外抑制活性进行评估，以部分验证其药理活性。HPLC 用于测定含量。
根据基于网络的分析，从 BMCT 网络中确定了 9 种具有重要意义的化合物，它们是 DS-CX 的 QC 标志物，包括丹参酮 I、丹参酮 IIA、隐丹参酮、丹酚酸 B、阿魏酸、丹酚酸 A、迷迭香酸、绿原酸和松柏醇阿魏酸酯。酶抑制试验部分验证了丹参酮 I 和丹参酮 IIA 的活性。化学分析表明，这 9 种标志物化合物是草药对的主要成分。
本研究鉴定了 DS-CX 草药对的基于活性的 QC 标志物，并提供了一种可用于其他草药、草药对或配方的 QC 的新方法。
