NDUFS3 alleviates oxidative stress and ferroptosis in sepsis induced acute kidney injury through AMPK pathway

• Published in: International immunopharmacology 2024-12, Vol.143 (Pt 2), p.113393, Article 113393
• Main Authors: Wang, YuChen, Lv, WuYang, Ma, XiaoTong, Diao, RuXue, Luo, XiaoXiao, Shen, QiuLing, Xu, MingYu, Yin, MengJiao, Jin, YingYu
• Format: Article
• Language: English
• Subjects: Acute Kidney Injury - metabolism; Acute Kidney Injury - pathology; AMP-Activated Protein Kinases - metabolism; Animals; Disease Models, Animal; Ferroptosis; Humans; Kidney - metabolism; Kidney - pathology; Lipopolysaccharides; Male; Mitochondria - metabolism; NADH Dehydrogenase - genetics; NADH Dehydrogenase - metabolism; Oxidative Stress; Rats; Rats, Sprague-Dawley; Sepsis - complications; Sepsis - metabolism; Signal Transduction
• ISSN: 1567-5769; 1878-1705; 1878-1705
• DOI: 10.1016/j.intimp.2024.113393

[Display omitted] •We found for the first time that NDUFS3 expression was downregulated in both in vitro and in vivo models of SI-AKI.•We found for the first time that NDUFS3 can inhibit LPS-induced ferroptosis, mitochondrial damage and oxidative stress in vivo and in vitro.•We found for the first time that NDUFS3 can regulate ferroptosis through AMPK pathway. In recent years, ferroptosis has been found to play an important role in various acute kidney injury (AKI). However, relatively little research has been conducted on sepsis-induced acute kidney injury (SI-AKI). As an important trigger of ferroptosis, how mitochondrial damage plays a regulatory role in SI-AKI is still unclear. To explore the potential relationship between mitochondria and ferroptosis, we established a SI-AKI rat model by intraperitoneal injection of lipopolysaccharide (LPS). Transcriptome sequencing was used to detect changes in gene transcription levels in the control group, LPS 3 h group, LPS 6 h group and LPS 12 h group. The severity of kidney injury was determined based on serum creatinine (CRE), blood urea nitrogen (BUN), tissue HE staining, TUNEL staining and inflammatory factor levels. Cytoscape software was utilized to screen several mitochondria-related HUB genes, and NADH dehydrogenase [ubiquinone] ferrithionein 3 (NDUFS3) was selected for subsequent validation due to its novelty and feasibility. qRT-PCR, Western blot was employed to evaluate the expression of NDUFS3 in kidney tissues. GO enrichment analysis revealed that up-regulated genes in the LPS 12 h group were enriched in several cell death terms while down-regulated genes were enriched in lipid metabolic process and oxidation–reduction progress terms. Furthermore, Western blot, IHC, MDA, GSH and iron content levels were used to assess ferroptosis in the kidney tissue of the SI-AKI rats, dihydroethidium (DHE) assay and ATP kit were used to assess mitochondrial ROS levels and mitochondrial function. To further validate the function of NDUFS3, we constructed overexpression rats using hydrodynamic method by tail vein injection of pc DNA3.1-NDUFS3 overexpression plasmid. we utilized LPS to stimulate HK-2 cells and establish an in vitro model. We then overexpressed NDUFS3 using pcDNA 3.1. The overexpression of NDUFS3 was found to inhibit LPS-induced ferroptosis and mitochondrial damage in HK-2 cells, as evidenced by Western blot, MDA, GSH, divalent iron, ROS levels, Mitosox red, ATP content and transmission electron micros