cell culture human kidney proximal tubular epithelial cells  (ATCC)

Journal: Cell Death Discovery

Article Title: IL‑1 receptor antagonism attenuates renal fibrosis via RNF182‑driven MFN2 destabilization and mitochondrial dysfunction

doi: 10.1038/s41420-025-02929-4

Figure Lengend Snippet: In the context of kidney injury, pro-inflammatory stimuli upregulate the expression of the Interleukin-1 Receptor (IL-1R) and its downstream effector, TGF-β1, in renal tubular epithelial cells. This leads to increased expression of the E3 ubiquitin ligase RNF182. RNF182, in turn, forms a complex with the mitochondrial fusion protein Mitofusin 2 (MFN2), targeting it for ubiquitination and subsequent proteasomal degradation. The depletion of MFN2 results in mitochondrial dysfunction, characterized by excessive reactive oxygen species (ROS) production and reduced ATP synthesis, which drives the progression of renal fibrosis. Therapeutic administration of recombinant human IL-1 receptor antagonist (rhIL-1Ra) blocks IL-1R signaling. This intervention suppresses the downstream upregulation of RNF182, thereby reducing MFN2 ubiquitination and degradation. The stabilization of MFN2 protein preserves mitochondrial function, mitigates oxidative stress, and restores cellular energy production, ultimately exerting a potent anti-fibrotic effect and protecting kidney tissue.

Article Snippet: Human kidney proximal tubular epithelial cells (HK-2, ATCC CRL-2190) were cultured in DMEM/F-12 medium (Gibco, Cat# 11330032) supplemented with 10% fetal bovine serum (FBS) (Gibco, Cat# 10270106) and 1% penicillin-streptomycin (Gibco, Cat# 15140122).

Techniques: Expressing, Ubiquitin Proteomics, Recombinant

Journal: Kidney Diseases

Article Title: CXCL6 Orchestrates Macrophage-Driven Inflammation in Diabetic Kidney Disease and Represents a Druggable Target

doi: 10.1159/000548806

Figure Lengend Snippet: Flow diagram of the study design. A comprehensive overview of the study workflow, illustrating the integration of bioinformatics analysis, experimental validation, and therapeutic exploration to investigate the pathogenesis of diabetic kidney disease (DKD). Gene expression datasets ( GSE30529 , GSE47184 , GSE175759 , and GSE142025 ) were analyzed to identify differentially expressed genes (DEGs) and key modules associated with DKD progression using weighted gene correlation network analysis (WGCNA). KEGG pathway enrichment analysis highlighted significant signaling pathways, and machine-learning approaches (LASSO, SVM, and RF) identified CXCL6 as the hub gene. Single-cell RNA sequencing (scRNA-seq) data further localized CXCL6 expression to proximal tubular epithelial cells in the kidney. Experimental validation was conducted using renal tissues from DKD patients and mice, as well as in vitro studies with HG-stimulated HK-2 cells and THP-1 macrophages to confirm CXCL6’s role in macrophage recruitment and inflammatory cytokine release. Virtual screening and molecular docking identified salvianolic acid B as a CXCL6-targeting inhibitor, which was shown to mitigate inflammation and macrophage migration in in vitro studies. This integrative approach bridges bioinformatics and experimental data, highlighting CXCL6’s role in DKD and its therapeutic potential.

Article Snippet: Human kidney tubular epithelial cells (HK-2) were obtained from ATCC (Shanghai, China).

Techniques: Biomarker Discovery, Gene Expression, Protein-Protein interactions, RNA Sequencing, Expressing, In Vitro, Migration

Journal: Kidney Diseases

Article Title: CXCL6 Orchestrates Macrophage-Driven Inflammation in Diabetic Kidney Disease and Represents a Druggable Target

doi: 10.1159/000548806

Figure Lengend Snippet: GSE142025 dataset identifies key modules associated with advanced DKD via WGCNA. a Correlation between module eigengenes and clinical traits of DKD. The turquoise module was closely related to advanced DKD. b The top 20 of significant KEGG pathways in turquoise module. c The expression level of CXCL6 in advanced DKD was significantly increased in GSE142025 database (*** p < 0.001). The mRNA expression level of CXCL6 was negatively correlated with eGFR in both the GSE30529 ( d ) and GSE175759 ( e ) databases. f Relative expression level of CXCL6 in the various cell types between healthy controls and DKD from the KIT website. g The protein expression of CXCL6 and proximal tubular epithelial marker (AQP1) in human kidney tissues of DKD and healthy control kidney using double immunofluorescence staining. h Representative CXCL6 immunohistochemical staining in healthy control kidney and renal biopsies of human DKD. Data were represented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. healthy control.

Article Snippet: Human kidney tubular epithelial cells (HK-2) were obtained from ATCC (Shanghai, China).

Techniques: Expressing, Marker, Control, Double Immunofluorescence Staining, Immunohistochemical staining, Staining