Core Constituents of

purposeAs a traditional herb product, the root of

methodsA successively partitioned extract of JQG, bioactive partition screening in vitro and in vivo, qualitative analysis, bioinformatic analysis, molecular docking, and mechanism validation were used in this study. The partitioned extract was used to obtain the bioactive partition, while in vitro anti-inflammatory effects and in vivo anti-arthritis effects in adjuvant-induced arthritis (AIA) rats were applied to screen the bioactive partition with the best efficacy. Qualitative analysis was used to identify bioactive constituents. Bioinformatic analysis was used to explore the potential mechanism for RA treatment. Molecular docking and immunofluorescence were used to validate the underlying mechanism.

resultsAfter successively partitioning extract and bioactive partition screening, ethyl acetate extract (EAE) yielded the best anti-inflammatory effects in vitro and in vivo among JQG extracts. By ultra-performance liquid chromatography (UPLC) coupled with Orbitrap mass spectrometry, a total of 58 constituents were identified in EAE, and 17 constituents were regarded as the core constituents based on their oral bioavailability and drug-like properties. The nuclear factor kappa B (NF-κB) signal pathway was screened as the core pathway of core constituents for RA treatment based on bioinformatic analysis, and the core constituents showed good ligand-receptor binding activity to NF-κB P65. In vitro study demonstrated that EAE could significantly reduce NF-κB P65 transfer from the cytoplasm to the nucleus.

conclusionOur study suggested that the therapeutic efficacy of JQG for RA treatment could be derived from negative regulation of the NF-κB pathway, and EAE of JQG could represent a promising herb product for RA treatment that deserves further development.