Integrated bioinformatics and experimental analysis of mitochondrial-associated membrane function and mechanism in acute respiratory distress syndrome​​.

Acute respiratory distress syndrome (ARDS) is a life-threatening lung condition characterized by severe inflammation, immune dysregulation, and oxidative stress, leading to high mortality (30-40%). This study explores the involvement of MAM-related genes in ARDS pathogenesis through bioinformatics and experimental validation. Publicly available RNA-sequencing data from ARDS and control samples were analyzed to identify differentially expressed genes (DEGs). Functional enrichment, gene set variation analysis (GSVA), and weighted gene co-expression network analysis (WGCNA) were performed to explore pathway alterations and hub gene interactions. Immune cell infiltration analysis was conducted using CIBERSORT. Candidate MAM-related genes were validated in a Poly I: C-induced ARDS mouse model and MLE-12 murine lung epithelial cells. The mouse model was assessed for lung histopathology, wet-to-dry lung weight ratio, bronchoalveolar lavage fluid (BALF) inflammatory cytokine levels (IL-1β and TNF-α), and lung injury scores. MLE-12 cells were treated with Poly I: C, and cell viability, lactate dehydrogenase (LDH) release, and apoptosis were evaluated. Protein-protein interaction (PPI) network analysis and drug prediction were used to identify potential therapeutic targets. A total of 3152 DEGs including 1549 upregulated and 1603 downregulated were identified in ARDS samples. Pathway analysis revealed autophagy suppression and immune activation, with 14 immune cell types significantly elevated in ARDS patients. Experimental validation confirmed that Poly I: C-induced ARDS mice exhibited severe lung injury and increased inflammatory reaction, while Poly I: C-treated MLE-12 cells showed increased cytotoxicity and LDH release. ZMAT2 and HBB were identified as key MAM-related hub genes, with ZMAT2 positively associated with disease progression and HBB negatively correlating with lung injury severity. Drug prediction analysis identified 29 pharmacological agents interacting with HBB, suggesting therapeutic potential. This study identifies ZMAT2 and HBB as key MAM-related genes contributing to ARDS pathogenesis, with potential diagnostic and therapeutic applications. The integration of bioinformatics with in vivo and in vitro validation provides novel insights into ARDS molecular mechanisms. Further clinical studies are needed to explore their translational relevance.