Buyang Huanwu Decoction attenuates vascular aging by suppressing the pathway of neutrophil extracellular trap formation via modulation of the HMGB1/TLR4/p38 signaling pathway.

BHD contains 23 major chemical constituents identified in this study.

• Chemical constituent identification was performed as part of characterizing the BHD formula.
• BHD is described as a classic traditional Chinese medicine formula widely used for treating cardio-cerebrovascular diseases.
• Identification of constituents was a prerequisite step before mechanistic investigation.

BHD ameliorated aging phenotypes in a D-galactose-induced aging mouse model.

• The D-galactose-induced aging mouse model was used as the primary in vivo experimental system.
• BHD treatment significantly alleviated aortic structural degeneration.
• BHD reduced oxidative stress and inflammatory cytokine levels in treated animals.
• BHD downregulated key senescence markers including p16 and p21.

Integrated multi-omics analysis identified NET suppression as a primary mechanism underlying the anti-aging benefits of BHD.

• The study combined transcriptomics, network pharmacology, and molecular biology methods.
• Multi-omics analysis focused on neutrophils as a central cell type.
• NET formation pathway was implicated as a core process in vascular aging acceleration.

BHD inhibits NETosis by modulating the HMGB1/TLR4/p38 signaling pathway both in vivo and in vitro.

• Both in vivo (D-galactose mouse model) and in vitro experiments confirmed BHD's inhibition of NETosis.
• BHD modulation of this pathway led to reduced expression of critical NET components.
• The HMGB1/TLR4/p38 cascade was established as the mechanistic link between BHD and NET suppression.

HMGB1 overexpression partially reversed the inhibitory effects of BHD on NETosis, establishing HMGB1 as a key effector molecule.

• HMGB1 overexpression experiments were used to validate the mechanistic role of HMGB1.
• Partial reversal of BHD's inhibitory effects upon HMGB1 overexpression confirmed HMGB1 as a key downstream effector.
• This finding positioned HMGB1 at the top of the signaling cascade targeted by BHD.

This study is the first to unveil a mechanism whereby BHD alleviates vascular aging through inhibition of the HMGB1-TLR4-p38-NETs cascade.

• Prior to this study, whether BHD acted through modulating NET-driven vascular aging was unknown.
• The authors describe this as providing 'a novel immunomodulatory perspective on BHD.'
• The findings highlight BHD's potential as 'a holistic therapeutic strategy against vascular aging.'

Excessive formation of neutrophil extracellular traps (NETs) is identified as a core immune-inflammatory process accelerating vascular aging.

• Vascular aging is described as a significant driver of age-related cardiovascular diseases.
• The immune-inflammatory response driven by NETs is characterized as a core process in vascular aging progression.
• This background provided the rationale for investigating NETs as a target of BHD action.