Integrated serum pharmacochemistry and network pharmacology reveal the mechanisms of Baoyuan decoction formula granules against spleen deficiency syndrome.

Abstract

INTRODUCTION: The aim of this study is to investigate the active material basis and action mechanism of BYD in spleen deficiency. METHODS: First, to study the therapeutic effect of BYD on spleen deficiency, a spleen deficiency mice model was established by injected reserpine. Then, the UPLC-QTOF-MS/MS was used to analyze the chemical composition characteristics of BYD formula particles and the blood-absorbed constituents. Next, the correlation network between the blood-absorbed constituents of BYD formula particles and spleen deficiency was established, and the action mechanism of the key active components was confirmed by KEGG enrichment analysis. Finally, the mechanism of action was verified by Western blot and Real time Quantitative PCR experiments. Apart from that, the in vivo safety of BYD formula particles was evaluated by toxicological experiments. RESULT: BYD formula particles significantly improved weight loss, restored immune organ index, and inhibited inflammation in mice with spleen deficiency. 249 active ingredients were identified in BYD formula granules, of which 47 could be absorbed by bloodstream such as Liquiritigenin, Wogonoside, Isoliquiritigenin and Oroxylin A-7-O-β-D-glucuronide, they were considered as the key active ingredients in BYD treatment for spleen deficiency. Network pharmacology analysis found that BYD mainly treated spleen deficiency by regulating apoptosis and angiogenesis. BYD upregulated Bcl-2, downregulated Bax and caspase3 expression, which inhibited immune cell apoptosis. The expression of vascular growth factor VEGF was significantly upregulated, that is beneficial to alleviate symptoms of spleen deficiency. In addition, toxicological experiments showed that BYD was safe for the internal organs and blood system of mice. CONCLUSIONS: This study validated the efficacy and safety of BYD formula granules in the treatment of spleen deficiency, and preliminarily investigated the mechanism of BYD in improving spleen deficiency by inhibiting apoptosis and promoting angiogenesis. That provide reference for the further clinical application of BYD formula granules.