Intervention of Sodium Hydrosulfide on Cuproptosis of RatAlveolar Epithelial Type II Cells Induced by Hypoxia/Reoxygenation
CHENG Yuan1#, LUO Zhenzhen1#, CHEN Jihao2#, HAN Congmei1, LIN Zhiyi3, HUANG Man1, DAI Yongyue1,CHEN Linglong2, CHEN Xinguo2, WANG Wantie1*
This study aims to investigate the effects and potential mechanisms of NaHS (sodium hydrosulfide), an exogenous hydrogen sulfide donor, on H/R (hypoxia/reoxygenation)-induced injury in rat ATII (alveolar epithelial type II cells), focusing on the role of cuproptosis-related molecular pathways. The RLE-6TN rat ATII cell linewas used to establish an H/R injury model. Cell viability was assessed by CCK-8 assay; intracellular Cu⁺ levels weremeasured using a fluorescent probe; protein expression levels of copper transporters and cuproptosis-related proteinswere determined by Western blot; mRNA expression levels of cuproptosis-related genes were quantified by RT-qPCR;mitochondrial membrane potential was evaluated by JC-1 staining; ATP content was measured by chemiluminescence; PDHA1 level was detected by ELISA; and citric acid content was quantified by the sulfosalicylic acid method.H/R treatment significantly suppressed ATII cell viability and induced oxidative stress. H/R also led to intracellularCu⁺ accumulation, upregulation of the copper importer CTR1, and downregulation of the copper exporters ATP7Aand ATP7B. Meanwhile, the cuproptosis-related proteins DLAT and DLST were significantly downregulated, whileFDX1 and LIAS were upregulated, accompanied by increased formation of DLAT oligomers. The expression levelsof the cuproptosis-related genes FDX1 and LIAS were upregulated, whereas DLAT and DLD were downregulated.In addition, H/R decreased mitochondrial membrane potential, ATP content, PDHA1 level and citric acid content.Pretreatment with 50 μmol/L NaHS significantly reversed all of the above changes. H/R activates cuproptosis-relatedmolecular pathways in rat ATII cells, accompanied by copper metabolic dysregulation, mitochondrial dysfunction anddecreased levels of metabolic intermediates. NaHS pretreatment alleviates H/R-induced ATII injury by restoring copper metabolic homeostasis, suppressing cuproptosis-related pathway activation and improving mitochondrial function.
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