Study on the Active Ingredients and Mechanism of Action of Zhenggu Zijin Wan Based on UPLC-Orbitrap Fusion Lumos Tribrid-MS, Network Pharmacology, and Experimental Validation

OBJECTIVE To predict the active ingredients and mechanism underlying the blood circulation-promoting and stasis-resolving effects of Zhenggu Zijin Wan using UPLC-Orbitrap Fusion Lumos Tribrid-MS, network pharmacology, and experimental validation.

METHODS  Firstly, UPLC-Orbitrap Fusion Lumos Tribrid-MS was employed to rapidly characterize the chemical components in Zhenggu Zijin Wan. Secondly, a “drug-component-target” network was constructed through network pharmacology research methods to obtain key targets and major active components. The protein-protein interaction(PPI) network was constructed by combining the String platform and CytoScape software. Pathway enrichment analysis was conducted through the MateScape database, and molecular docking verification was performed using DiscoVery Studio 4.5 software. Finally, an animal model of acute soft tissue injury was established, and pharmacodynamic studies were conducted using acute soft tissue injury scores and whole blood viscosity as pharmacodynamic indicators.

RESULTS A total of 67 compounds including flavonoids, alkaloids, organic acids, and coumarins were identified in Zhenggu Zijin Wan. Among them, emodin, ligustilide, cinnamic acid, salicylic acid, aloe emodin, and others might be the main active ingredients for promoting blood circulation and removing blood stasis in Zhenggu Zijin Wan. PPI network analysis revealed 26 core targets such as TNF, ALB, AKT1. KEGG enrichment analysis showed that Zhenggu Zijin Wan mainly regulated TNF, PI3K-Akt, NF-κB signaling pathway played a role in promoting blood circulation and resolving stasis. And molecular docking results showed that the main active ingredients of Zhenggu Zijin Wan bound well with key targets. The pharmacodynamic results indicated that Zhenggu Zijin Wan could significantly reduce the whole blood viscosity in rats with acute soft tissue injury.

CONCLUSION The study preliminarily clarifies the active ingredients and mechanism of action of Zhenggu Zijin Wan in promoting blood circulation and resolving blood stasis, and indicates that it may exert the effect of promoting blood circulation and resolving blood stasis together by acting on multiple targets and pathways, providing reference for its future in-depth research.