Anticoagulant active ingredients identification of total saponin extraction of different Panax medicinal plants based on grey relational analysis combined with UPLC-MS and molecular docking.
ETHNOPHARMACOLOGICAL RELEVANCE
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?


AIM OF THE STUDY
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.


MATERIALS AND METHODS
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.


RESULTS
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.


CONCLUSION
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.
['Ren|Yongshen|Y|', 'Ai|Jiao|J|', 'Liu|Xinqiao|X|', 'Liang|Shuai|S|', 'Zheng|Yao|Y|', 'Deng|Xin|X|', 'Li|Yan|Y|', 'Wang|Jing|J|', 'Deng|Xukun|X|', 'Chen|Lin-Lin|LL|']
D000925:Anticoagulants D001777:Blood Coagulation D001780:Blood Coagulation Tests D002851:Chromatography, High Pressure Liquid D006801:Humans D062105:Molecular Docking Simulation D005894:Panax D010936:Plant Extracts D010946:Plants, Medicinal D012503:Saponins D015398:Signal Transduction D021241:Spectrometry, Mass, Electrospray Ionization D013329:Structure-Activity Relationship
UPLC-MS Molecular docking Grey correlation analysis Network pharmacology Panax ginseng total saponin anticoagulant activity