Identifying Baicalein as a Key Bioactive Compound in XueBiJing Targeting KEAP1: Implications for Antioxidant Effects

Area of Science:

• Pharmacology
• Molecular Biology
• Immunology

Background:

• XueBiJing injection (XBJ) exhibits multi-target effects like immunomodulation and antioxidation, beneficial for severe infections such as sepsis and COVID-19.
• The precise molecular targets and mechanisms of XBJ remain incompletely understood, necessitating further research.

Purpose of the Study:

• To identify key bioactive compounds within XBJ.
• To elucidate the molecular targets and mechanisms of action for these compounds.

Main Methods:

• Zebrafish models were used to assess anti-inflammatory and antioxidant properties.
• RNA sequencing and network analysis identified differentially expressed genes (DEGs) and molecular targets.
• Network pharmacology, molecular docking, kinetic simulations, CETSA-WB, SPR, and gene silencing experiments were employed to validate compound-target interactions and pathway activation.

Main Results:

• XBJ demonstrated dose-dependent anti-inflammatory effects by reducing neutrophil and macrophage counts in zebrafish.
• Transcriptomic analysis revealed differential expression in the KEAP1/NRF2 pathway, with KEAP1 identified as a central target.
• Baicalein was confirmed as a key active compound that directly binds to KEAP1, activating the KEAP1/NRF2 pathway, as evidenced by multiple validation techniques.

Conclusions:

• Baicalein is a critical bioactive component of XBJ responsible for its antioxidant and anti-inflammatory activities.
• The mechanism involves direct binding of baicalein to KEAP1, leading to the activation of the KEAP1/NRF2 pathway.
• This study provides novel insights into the therapeutic mechanisms of XBJ for severe infections.