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Downregulated RSAD2 Attenuates the Apoptosis of LPS-Stimulated RAW264.7 via NF-κB Signaling Pathway

Year: 2025

Type: article

Abstract: Background: This study aimed to investigate the role of Radical S-adenosyl Methionine Domain-Containing 2 (RSAD2) in regulating the apoptosis of LPS-stimulated RAW264.7 macrophages via the NF-κB signaling pathway. Methods: Differentially expressed genes in LPS-stimulated macrophages were identified using a gene expression dataset from the Gene Expression Omnibus (GEO) database and analyzed with R software. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analyses were performed to predict the biological functions of the identified genes. Key genes involved in NF-κB-mediated apoptosis regulation were selected for further investigation. The expression levels of Bcl-2, cleaved caspase-3, and NF-κB p65 were assessed by Western blotting. TUNEL staining was used to evaluate apoptosis. Results: RSAD2 knockdown significantly improved cell viability and reduced apoptosis in LPS-stimulated RAW264.7 cells. Downregulation of RSAD2 increased Bcl-2 expression and inhibited cleaved caspase-3 activity, thus inhibiting apoptosis. Mechanistically, the downregulation of RSAD2 suppressed the NF-κB signaling pathway in vitro. Treatment with phorbol 12-myristate 13-acetate (PMA), an NF-κB agonist, reversed the protective effects of RSAD2 knockdown in LPS-stimulated RAW264.7 cells. Conclusion: Our findings suggested that RSAD2 knockdown alleviated LPS-induced apoptosis in RAW264.7 macrophages by suppressing the NF-κB signaling pathway, highlighting RSAD2 as a potential therapeutic target for sepsis-related macrophage dysfunction.

Source: Journal of Inflammation Research

DOI: 10.2147/jir.s535225

Authors: Zhen Qi, Kongli Lu, Mengxi Huang, Haixia Wang, Rongheng Liao, Ri Tang