Sesamin exerts protective effects against diquat-induced acute kidney injury by inhibiting oxidative stress, apoptosis and autophagy: A key role for the SIRT5/FOXO3a signaling pathway.

CSB6B attenuates renal inflammation and fibrosis by inhibiting the activation of NLRP3 inflammasome through the NLRP3/Caspase-1/GSDMD/IL-1β signaling pathway.

Sepsis-related acute kidney injury (S-AKI) is a severe condition characterized by rapid onset and high mortality. Thus, identifying effective treatments for S-AKI is of critical importance. Lipopolysaccharide (LPS) was used to activate HK-2 cells to mimic S-AKI in vitro. Lentiviral transfection was performed to knock down C-type lectin domain family 5 member A (CLEC5A) expression, and protein immunoblotting was used to detect changes in CLEC5A expression. Cell damage was evaluated using the cell counting kit-8 (CCK-8) and lactate dehydrogenase (LDH) kit, cellular inflammatory factor levels were determined using the enzyme-linked immunosorbent assay (ELISA), and oxidative stress signs were detected using the kit. Western blotting was used to detect the expression of NF-κB/NLRP3 (NLR family, pyrin domain-containing protein 3) pathway, and NF-κB activator was used to detect whether knockdown of CLEC5A acts through the NF-κB/NLRP3 pathway. LPS stimulated the expression of CLEC5A in HK-2 cells. Knockdown of CLEC5A could inhibit the LPS-induced decrease in HK-2 cell viability and increased LDH release. Knockdown of CLEC5A could inhibit the LPS-induced increase in HK-2 cell release of inflammatory factors. Knockdown of CLEC5A could inhibit LPS-induced oxidative stress. CLEC5A knockdown can prevent the NF-κB/NLRP3 signaling pathway from being activated, and NF-κB activation can undo the effects of CLEC5A knockdown. Knockdown of CLEC5A can ameliorate renal tubular damage and lessen inflammation and oxidative stress via reducing NF-κB/NLRP3 activation.

Harmine attenuates renal fibrosis via Twist1 suppression: A novel anti-fibrotic strategy for chronic kidney disease with efficacy/safety profiling.

Gentisic acid inhibits renal fibrosis by reprogramming glycolytic metabolism and regulating H3K18la.

Background: Acute kidney injury (AKI) frequently progresses to chronic kidney disease (CKD) through the AKI-to-CKD transition; however, effective treatment strategies remain challenging due to the complex and multifactorial pathophysiology of this process. This study aims to develop a multifunctional nanoplatform for kidney-specific targeting, reactive oxygen species (ROS) scavenging, and anti-fibrotic drug delivery to mitigate AKI-to-CKD progression. Methods: Reduced graphene oxide (rGO) was conjugated with hyaluronic acid (HA) to form HA/rGO nanoparticles, enabling CD44-mediated renal targeting and ROS-responsive drug release. Paricalcitol, a hydrophobic anti-fibrotic agent, was loaded onto HA/rGO to form P/HA/rGO. The physicochemical characteristics, ROS-scavenging capacity, and oxidative stress-responsive drug release were evaluated. In vitro cytoprotection was assessed using HK-2 cells under oxidative stress. In vivo studies using ischemia/reperfusion (IR) injury mouse models assessed biodistribution, renal targeting, and therapeutic efficacy after systemic administration of P/HA/rGO. Results: HA/rGO nanoparticles demonstrated potent antioxidant activity and significantly protected HK-2 cells from ROS-induced cytotoxicity. P/HA/rGO exhibited a high paricalcitol loading efficiency (93%) and released 26% of the drug over 30 days under oxidative conditions. P/HA/rGO selectively accumulated in IR-injured kidneys via HA-CD44 interactions, decreased serum NGAL and cystatin C levels, and effectively attenuated tubular injury, fibrosis, inflammation, and apoptosis compared to vehicle-treated controls. Conclusion: The P/HA/rGO nanoplatform enables kidney-targeted delivery of paricalcitol with ROS-scavenging and ROS-responsive release properties, providing a promising therapeutic strategy to suppress the AKI-to-CKD transition via integrated targeting and microenvironment-responsive therapy.

TCM compatibility-driven mechanism of Shenshuaining (SSN) capsules against renal fibrosis: Integrated metabolomic and network analyses.

The AMPK/ULK1/autophagy pathway: FGF23's weapon against GSDME-mediated pyroptosis in folic acid-induced acute kidney injury.

Dahuang Gancao decoction suppresses ferroptosis via SIRT3-mediated NRF2 deacetylation in acute kidney injury.

Targeting PIM1 ameliorates sepsis-associated acute kidney injury through inhibiting ferroptosis.

Apelin-13 alleviated ischemia reperfusion injury via KLF2/NF-κB signaling pathway after kidney transplantation.

Integrative network toxicology and experimental validation reveal novel molecular mediators of omeprazole-induced nephrotoxicity.

Background Nephrolithiasis is a significant health issue causing pain, hypertension, and chronic kidney disease. Phillyrin, a bioactive compound from Forsythia suspensa, may offer therapeutic benefits against these conditions. The study was focused on phillyrin efficacy on alleviating kidney stone formation and associated damage. Methods HK-2 cells were exposed to calcium oxalate (CaOx) to generate an in vitro kidney cell injury model. The kidney stones were induced in rats by 1% ethylene glycol and 2% ammonium chloride. Cell viability was assessed by CCK-8 assay. Cell apoptosis was detected by JC-1 mitochondrial membrane potential assay. ROS and MDA contents and the activity of SOD and GSH-Px were assayed using matched assay kits. Differentially expressed genes (DEGs) were identified by RNA sequencing (RNA-seq). Results Phillyrin significantly reduced CaOx-induced apoptosis and oxidative stress in HK-2 cells. Moreover, phillyrin repressed oxidative stress and renal crystal deposition in model rats of kidney stones. RNA-seq revealed DEGs in rat kidneys post-phillyrin treatment, suggesting involvement in key pathways. Phillyrin activated the PPARγ signaling pathway in kidney stone induced rats and CaOx-exposed HK-2 cells. Furthermore, the PPARγ antagonist GW9662 exerted a counteracting impact on phillyrin-mediated anti-apoptosis and anti-oxidative effects in CaOx-exposed HK-2 cells. Conclusion The present study demonstrated that the potential efficacy of phillyrin on alleviating CaOx kidney stone formation and associated oxidative stress. PPARγ signaling is the possible mechanism of action of phillyrin in controlling nephrolithiasis-triggered apoptosis.

Purpose This study aims to investigate the protective effects and mechanisms of 20S-protopanaxadiol (PPD), a key component derived from ginseng folium, against diabetic nephropathy (DN). Methods Ingredients of ginseng folium were collected and screened using the TCMSP database. Therapeutic target genes for ginseng folium against DN were obtained, and protein-protein interaction (PPI) networks were constructed. Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichments were conducted on these targets. Furthermore, the results of network pharmacology were validated via molecular docking. Subsequently,the effects of PPD on TGF-β1/Smads signaling pathway were assessed in db/db mice and HK-2 cells using biochemical assays and Western blotting. Results A total of 246 target genes for ginseng folium against DN were identified. KEGG analysis revealed significant enrichment in AGE-RAGE signaling in diabetic complications, PI3K-AKT, MAPK, and TNF pathways. PPD intragastric administration improved renal function in db/db mice and significantly reduced the expression of TGF-β1, α-SMA, p-Smad3, and Smad4 in renal tissue (p < 0.05). Furthermore, PPD decreased the expression of TGF-β1, p-Smad3, and Smad4 in high glucose-induced HK-2 cells. Conclusion PPD potentially alleviates renal fibrosis in db/db mice and HK-2 cells by inhibiting the TGF-β1/Smads pathway, thereby delaying DN progression.

Aims This study aimed to elucidate the role and function of roxadustat, a small molecule hypoxia-inducible factor (HIF) stabilizer, on cisplatin-induced acute kidney injury (AKI) in vivo and in vitro. Methods We investigated the effects of roxadustat (10 mg/kg) in a cisplatin-induced AKI model in mice. After a 48-hour pretreatment with the intraperitoneal injection of roxadustat, cisplatin was administered at a dose of 20 mg/kg by intraperitoneal injection. We also examined the role of roxadustat (10 μM) on HK-2 cells stimulated by cisplatin (20 μg/mL). Results Pretreatment with roxadustat significantly improved kidney function in the AKI model, as evidenced by reductions of Scr and BUN, and alleviated cisplatin-induced kidney morphology lesions. Roxadustat also remarkably decreased the expression of KIM-1 and NGAL. Furthermore, roxadustat significantly abolished the expression of Bax and cleaved caspase 3, two apoptosis marker proteins. Mechanistically, our study demonstrated that roxadustat could alleviate the deficiency of fatty acid oxidation (FAO) induced by cisplatin, as evidenced by the improvement of lipid accumulation and metabolic disorders in the roxadustat-treated group. Moreover, roxadustat effectively blocked mitochondrial damage, as evidenced by reduced mitochondrial swelling and fragmentation in the roxadustat-treated group. Simultaneously, roxadustat also partially rescued the expression of mitochondrial fusion proteins, including OPA1, MFN1, and MFN2. In vitro, roxadustat also effectively alleviate injury, apoptotic responses, FAO disorders, mitochondrial damage caused by cisplatin. Conclusion We demonstrated that roxadustat exerts a protective role in cisplatin-treated mice and HK-2 cells, which might be due to amelioration of FAO deficiency and suppression of mitochondrial damage.

Objective O-linked N-acetylglucosamine (O-GlcNAc) glycosylation represents a prevalent post-translational modification of proteins. Accumulating evidence indicates that dysregulated O-GlcNAcylation can induce glucose toxicity and plays critical roles in the pathogenesis of diabetic nephropathy (DN). Methods Human kidney proximal tubular epithelial HK-2 cells were exposed to high glucose (HG) conditions to establish DN cellular models. Co-immunoprecipitation (Co-IP) combined with western blot analysis was employed to assess the expression levels of O-GlcNAc, O-GlcNAc transferase (OGT), O-GlcNAcase (OGA), ACSL4, and ferroptosis-associated proteins. Cell viability was evaluated using the CCK-8 assay, while cell death was analyzed through DAPI/propidium iodide (PI) double staining. Ferroptosis was assessed by measuring intracellular iron accumulation, glutathione (GSH) content, and reactive oxygen species (ROS) levels. Additionally, in vivo experiments were conducted using male C57BL/6J mice divided into four groups: control, DN, DN+shOGT, and DN+shNC. DN was induced by a high-fat diet (HFD) followed by streptozotocin (STZ) injection. OGT was specifically knocked down in the kidneys using AAV-shOGT. Kidney tissues were analyzed for pathological changes, O-GlcNAcylation levels, and ferroptosis markers. Results Our findings demonstrated that O-GlcNAc and OGT expressions were significantly upregulated in HG-treated HK-2 cells. OGT knockdown effectively attenuated HG-induced ferroptosis. Importantly, ACSL4 protein levels exhibited strong positive correlation with OGT expression. Subsequent investigation revealed direct interaction between ACSL4 and OGT. O-GlcNAcylation modification was found to enhance ACSL4 protein stability. Moreover, overexpression of ACSL4 counteracted the protective effects of OGT knockdown against ferroptosis. Additionally, OGT knockdown reversed the high iron concentration, ROS, and MDA levels, and restored GSH and SOD levels in DN mice. O-GlcNAc, OGT, and ACSL4 levels, which were markedly increased in the DN group, were downregulated by OGT knockdown treatment in vivo. Conclusions Collectively, these results demonstrate that high glucose promotes OGT-mediated O-GlcNAcylation of ACSL4, thereby stabilizing this enzyme and facilitating ferroptosis progression in DN.

Background PAX2 is a key developmental gene, and its mutations are primarily associated with kidney and ocular anomalies, predominantly affecting children. This study aims to analyze the clinical manifestations and genetic characteristics of children with PAX2 mutations and to assess the functional impact of novel variants. Methods Clinical data were retrospectively reviewed in 10 children diagnosed with PAX2 mutations through whole-exome sequencing from a pediatric hereditary disease cohort database. AlphaFold 3 (AF3) was used for protein structural modeling. Novel variants were functionally assessed via HK-2 cell proliferation assays. Results The median age at initial presentation was 4.2 years (IQR 0–6.5). All 10 patients presented with proteinuria or microscopic hematuria. Nine had kidney dysplasia, and five progressed to stage 5 chronic kidney disease. Five patients had PAX2-related ocular abnormalities. Hepatic dysfunction and spermatic cord hydrocele were reported as potential novel phenotypes. A total of nine distinct PAX2 mutations were identified, including five novel variants. AF3 modeling revealed significant conformational disruptions in the novel variants, with root mean square deviation (RMSD) values ranging from 1.1 to 43.6 Å. Functional assays demonstrated that four novel variants significantly impaired the proliferative capacity of HK-2 cells. Conclusions This study characterizes five novel PAX2 variants with confirmed structural (RMSD 1.1–43.6 Å) and functional (HK-2 proliferation impairment) impacts, expanding both the mutational and phenotypic spectra in Chinese children. The findings highlight the association between PAX2 mutations and early-onset kidney disease with potential extrarenal involvement. Early genetic diagnosis and timely kidney-protective interventions are essential to improve outcomes.

Contrast-induced acute kidney injury (CI-AKI) is the third leading cause of AKI, but there are no effective preventive or therapeutic measures in clinical practice. Glycyrrhizin, a bioactive compound isolated from the Glycyrrhiza glabra L., exhibits anti-inflammatory effects; however, the effects and mechanisms of glycyrrhizin on CI-AKI remain unknown. In present study, the effects of glycyrrhizin on renal dysfunction and tissue damage were evaluated in CI-AKI rats and mice. And the mechanisms were further investigated in iohexol treated renal tubular epithelial cells. Molecular docking and network pharmacology were used to discover the binding targets of glycyrrhizin and identify potential pathogenic pathway. Gene knockout mice and gene silencing cells were used to detect whether glycyrrhizin alleviated CI-AKI through target proteins mediated pathway. Results showed that both pretreatment and co-treatment with glycyrrhizin could alleviate iohexol-induced renal dysfunction and pathological damage in vivo. Similarly, glycyrrhizin could improve iohexol-induced decrease in cell viability of both HK-2 cells and primary mice renal tubular epithelial cells. Mechanistically, glycyrrhizin could directly bind to the active site of HMGB1, then blocking iohexol-induced ferroptosis of renal tubular epithelial cells. HMGB1 silencing was able to inhibit overactivation of AMPK/Beclin-1 axis during CI-AKI, and iohexol-downregulated protein expressions of GPX4 and SLC7A11 were reversed in kidneys of AMPK knockout mice. Comparable results were obtained in vitro with AICAR treatment. Our study is the first to demonstrate that glycyrrhizin exerts both protective and therapeutic effect on CI-AKI by inhibiting tubular epithelial cell ferroptosis via HMGB1/AMPK/Beclin-1 axis, providing a potential choice for treating CI-AKI.

Background Emerging evidence underscores the critical involvement of ferroptosis in the pathophysiology of AKI. However, the role of ferroptosis-related genes (FRGs) in AKI remains insufficiently explored. This study sought to identify potential FRGs associated with AKI through bioinformatics approaches and experimental validation. Methods AKI-related datasets and FRGs were first collected. Differentially expressed FRGs linked to AKI were identified through analytical methods, followed by an examination of their biological functions. Diagnostic biomarkers were then selected using LASSO, RFE, and RF algorithms. Additionally, small pharmacological molecules associated with DE-FRGs were identified to explore the connection between DE-FRGs and AKI. qRT-PCR analysis revealed FAR1 expression in AKI, while Western blotting and IHC confirmed corresponding FAR1 protein changes in kidney tissues. TUNEL staining confirmed cell death in AKI. ROS production and ferroptosis markers were evaluated in FAR1-knockdown and FAR1-overexpressing HK-2 cells. Results A total of 106 DE-FRGs were identified, with functional enrichment analysis revealing strong associations with the MAPK and mTOR signaling pathways, as well as ferroptosis. Eight diagnostic biomarkers were selected using multiple algorithms, and their predictive accuracy was validated through ROC curve analysis. Furthermore, 13 pharmacological molecules were identified to establish a relationship between DE-FRGs and AKI. AKI renal tissue exhibited elevated cell death and reduced FAR1 expression. In vitro, FAR1 knockdown in HK-2 cells increased ROS and ferroptosis markers, while FAR1 overexpression rescued these phenotypes. Conclusion This study identified signaling pathways and small molecules associated with DE-FRGs in AKI. FAR1 was also identified as a potential diagnostic biomarker for AKI.

BackgroundUromodulin has been speculated as a protective factor for acute kidney injury (AKI) and chronic kidney disease (CKD), possibly based on the uromodulin-mediated cross-talk between thick ascending limbs and proximal tubules. Here, we explored the roles of uromodulin in the AKI-CKD transition.MethodsWild-type SD rats and UMOD−/− rats were subjected to the AKI-CKD transition models. Cisplatin-stimulated HK-2 cells were treated with or without uromodulin. The renal injury and cell damage were evaluated.ResultsUMOD−/− rats observed no spontaneous kidney injury and fibrosis. In two rat models of AKI-CKD transition, induced by repeated cisplatin nephrotoxicity and ischemia-reperfusion injury, UMOD deficiency exacerbated the kidney insufficiency and fibrosis along with the over-activation of EGFR signaling. In the kidneys treated with cisplatin repeatedly, immunofluorescence showed the translocation of uromodulin and its proximity to EGFR and coimmunoprecipitation experiments also verified their interaction. Molecular docking and microscale thermophoresis demonstrated a significant binding capacity of EGFR with uromodulin and its EGF-like domains. Supplying uromodulin to cisplatin-stimulated HK-2 cells decreased the fibrotic process and the EGFR abundance, associated with the elevated EGFR ubiquitylation. Recombinant EGF-like domains of uromodulin showed a similar binding capacity and EGFR suppression effect with uromodulin. Uromodulin-induced EGFR endocytosis was impaired by the EGFR kinase inhibitor or monoclonal antibody.ConclusionsCollectively, we discovered uromodulin may alleviate the progression of AKI-CKD transition via the EGFR suppression.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12964-025-02537-w.