Biochanin A alleviates endothelial cell senescence via epigenetic regulation of the HDAC1/H3K4me3/NFKB1 axis

BACKGROUND: Aging is a multifaceted physiological and pathological process, with endothelial cell senescence being a pivotal contributor to vascular aging. Biochanin A (BCA), a naturally occurring flavonoid, possesses known anti-inflammatory and antioxidant properties; however, its specific role and mechanism in combating endothelial senescence are not well defined. PURPOSE: This study aimed to investigate the potential anti-senescent effects of BCA on endothelial cells and aged vasculature, and to elucidate the underlying molecular mechanisms. STUDY DESIGN: The study employed a combination of in vitro cellular models and in vivo animal models to assess the effects of BCA on senescence. Mechanistic investigations were conducted to identify key molecular targets and pathways. METHODS: In vitro, an Angiotensin II (Ang II)-induced senescence model in human umbilical vein endothelial cells (HUVECs) was used. In vivo, naturally aged mice were treated with BCA. Senescence markers and SASP factors (e.g., IL-6, IL-8) were evaluated. RNA sequencing was performed to identify potential targets. Histone modification changes were analyzed through genomic binding site analysis, CUT&Tag assays, and chromatin immunoprecipitation quantitative PCR (ChIP-qPCR). Knockdown of histone deacetylase 1 (HDAC1) was achieved using siRNA to verify its necessity. RESULTS: BCA treatment effectively attenuated senescence markers and reduced senescence-associated secretory phenotype (SASP) levels in both Ang II-induced HUVECs and naturally aged mice. RNA sequencing identified HDAC1 as a key target. Mechanistically, BCA upregulated HDAC1, which subsequently suppressed the epigenetic aging marker H3K4me3. CUT&Tag and ChIP-qPCR analyses confirmed that this reduction in H3K4me3 occurred specifically at the promoter region of NFKB1 (p50), leading to its transcriptional downregulation. This inhibition of NFKB1 suppressed NF-κB signaling and the production of SASP factors. Crucially, HDAC1 knockdown completely abolished the anti-senescence benefits of BCA. CONCLUSION: Our findings demonstrate that Biochanin A alleviates endothelial and vascular senescence by targeting the HDAC1/H3K4me3/NFKB1 axis. This epigenetic mechanism inhibits NF-κB-mediated SASP production, positioning BCA as a promising natural epigenetic modulator to restore vascular homeostasis and mitigate age-related vascular dysfunction.