School of Pharmaceutical Science, South-Central University for Nationalities, Wuhan, Hubei, 430074, PR China.
Key Laboratory of Traditional Chinese Medicine Resource and Compound Prescription, Ministry of Education, Hubei University of Chinese Medicine, Wuhan 430065, PR China.
J Ethnopharmacol. 2020 Oct 5;260:112955. doi: 10.1016/j.jep.2020.112955. Epub 2020 May 16.
In traditional use, Panax medicinal plants (ginseng, red ginseng, notoginseng, Panax japonicus, and Panacis majoris rhizoma) have different bioactivities from each other, even under different dosages, but their chemical compositions are very similar; so the question is, what is the primary effective substance induced the different efficacy, and how to identify them from a group of chemical constituents?
The goal of this research was to provide a strategy to determine the effective substance in Panax genus medicinal plants responsible for the anticoagulant response.
This research used ultra-high performance liquid chromatography coupled with mass spectrometry (UPLC-MS) to analyze the saponin chemical compositions of different concentration ethanol extraction of notoginseng and the ginseng genus medical plant 75% ethanol extraction (Panax ginseng, red ginseng, Panax japonicus, Panacis majoris rhizome), and used four coagulation factors (PT, TT, APTT, Fib) to evaluate the anticoagulant activity of the extracts. Grey correlation analysis was applied to establish the spectral effect relationship and give the anticoagulant potency of different saponins. Network pharmacology and molecular docking were adopted to clarify and verify the possible mechanisms of anticoagulant action.
The results showed that the blood physiological regulation activities of Panax medicinal plants were different according to the solvent concentration, processing, species and dosage. Overall, the most suitable solvent for extraction of SQ was 75% ethanol; At low dosage (10-100 mg/mL), the anticoagulant effect of Panax medical plants was: ZJS > ZZS > SQ > RS > HS, and at high doses (100-1000 mg/mL) was: SQ > ZJS > ZZS > RS > HS. GRA and molecular docking results showed the contribution of some components (NG-R2, NG-Fc/G-Ra1/G-Ra2, G-Rc, G-Rk3, and G-Rh4) to the whole anticoagulant activity of the drug were increased, while the effect of CS-IVa was just decreased with the increase of dosage; the anticoagulant effect of G-Rg3 (the main anticoagulant component) is mainly related to the targets F2, AR, RHO, ACR, MB, GZMB, B2M, CA2, CAT, and PAPOLA.
This study determined the effective substance of anti-coagulation of ginseng genus herbal medicines and the regulation of different anticoagulant effects of TCM by changing various influencing conditions, including processing method, extraction method, and dose. It also provided an effective strategy for effective substances identification of multicomponent, multifunction, and multipurpose herbal medicine.
在传统用途中，人参属药用植物（人参、红参、三七、日本人参和 Panacis majoris rhizoma）彼此之间具有不同的生物活性，即使在不同剂量下也是如此，但它们的化学成分非常相似；因此，问题是，是什么主要有效物质引起了不同的疗效，以及如何从一组化学成分中识别它们？
本研究旨在提供一种策略，以确定人参属药用植物中负责抗凝反应的有效物质。
本研究使用超高效液相色谱-质谱联用（UPLC-MS）分析不同浓度乙醇提取的三七和人参属药用植物 75%乙醇提取物（人参、红参、日本人参、 Panacis majoris rhizome）的皂苷化学成分，并使用四种凝血因子（PT、TT、APTT、Fib）评估提取物的抗凝活性。采用灰色关联分析建立谱效关系，给出不同皂苷的抗凝效价。采用网络药理学和分子对接阐明并验证抗凝作用的可能机制。
结果表明，根据溶剂浓度、加工、种类和剂量的不同，人参属药用植物的血液生理调节活性也不同。总体而言，提取 SQ 最适宜的溶剂是 75%乙醇；在低剂量（10-100mg/mL）下，人参属药用植物的抗凝作用为：ZJS＞ZZS＞SQ＞RS＞HS，而在高剂量（100-1000mg/mL）下为：SQ＞ZJS＞ZZS＞RS＞HS。GRA 和分子对接结果表明，一些成分（NG-R2、NG-Fc/G-Ra1/G-Ra2、G-Rc、G-Rk3 和 G-Rh4）对药物整体抗凝活性的贡献增加，而 CS-IVa 的作用则随着剂量的增加而降低；G-Rg3（主要抗凝成分）的抗凝作用主要与 F2、AR、RHO、ACR、MB、GZMB、B2M、CA2、CAT 和 PAPOLA 等靶点有关。
本研究通过改变各种影响条件，包括加工方法、提取方法和剂量，确定了人参属草药的抗凝有效物质以及 TCM 对不同抗凝作用的调节，为多成分、多功能、多用途草药的有效物质鉴定提供了有效策略。
