model establishment human renal tubular epithelial cells hk 2  (Procell Inc)

Journal: Frontiers in Pharmacology

Article Title: Elucidating the therapeutic efficacy and mechanisms of arctigenin in ameliorating renal fibrosis: a combined transcriptomic and proteomic study

doi: 10.3389/fphar.2026.1796732

Figure Lengend Snippet: ATG ameliorates UUO-induced RF in rats by modulating the S100A8/A9/NOX/NF-κB signaling pathway, and additionally attenuates TGF-β1-induced fibrotic responses in HK-2 cells. (A–F) Relative mRNA expression levels of key genes (S100A8, S100A9, NOX2, NOX4, IκBα, and NF-κB p65) in the S100A8/A9/NOX/NF-κB pathway were measured via RT-qPCR in renal tissues. (G–L) Protein expression levels of S100A8, S100A9, NF-κB p65, and phosphorylated NF-κB p65 were detected using Western blot, including representative bands and quantitative analysis. (M) Cell viability of HK-2 cells under varying concentrations of ATG was assessed via CCK-8 assay. (N) Cell viability under different concentrations of PAQ was similarly evaluated using CCK-8. (O) Changes in HK-2 cell viability before and after drug treatment were determined via CCK-8. (P-R) mRNA expression levels of α-SMA, collagen I, and fibronectin in cells were quantified through RT-qPCR. (S–U) Western blot results and quantitative analysis of α-SMA and vimentin protein expression in cells. (V–W) Representative images of wound healing assays at 0 h and 24 h (scale bar = 100 μm) along with quantitative analysis of cell migration rates. Data are presented as mean ± SEM, n = 3 per group ( n = 6 per group for A-F), * p < 0.05, ** p < 0.01, *** p < 0.001, ns, no significant.

Article Snippet: Human renal cortical proximal tubular epithelial HK-2 cells (STCC10303P, Servicebio) were maintained in a humidified incubator (Thermo Fisher; MA, USA) at 37 °C with 5% CO 2 , and cultured in DMEM/F-12 medium (G4612) supplemented with 10% fetal bovine serum (G8003) and 1% penicillin-streptomycin (G4003).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Migration

Journal: Frontiers in Pharmacology

Article Title: Elucidating the therapeutic efficacy and mechanisms of arctigenin in ameliorating renal fibrosis: a combined transcriptomic and proteomic study

doi: 10.3389/fphar.2026.1796732

Figure Lengend Snippet: ATG alleviates TGF-β1-induced fibrosis in HK-2 cells by regulating the S100A8/A9/NOX/NF-κB signaling pathway. (A–F) Relative mRNA expression levels of key genes (S100A8, S100A9, NOX2, NOX4, IκBα, and NF-κB p65) in the S100A8/A9/NOX/NF-κB signaling pathway were measured in cells using RT-qPCR. (G–L) Relative protein expression levels of key proteins (S100A8, S100A9, NF-κB p65, and phosphorylated NF-κB p65) in the S100A8/A9/NOX/NF-κB signaling pathway were detected by Western blot, including representative band images and quantitative analysis. (M,N) Immunofluorescence images of NOX2 in cells and quantitative analysis of relative fluorescence intensity. (O,P) Immunofluorescence images of IKKβ in cells and quantitative analysis of relative fluorescence intensity. (Q,R) Immunofluorescence images of IκBα in cells and quantitative analysis of relative fluorescence intensity (scale bar = 50 μm). Data are presented as mean ± SEM, n = 3 per group, * p < 0.05, ** p < 0.01, *** p < 0.001, ns, no significant.

Article Snippet: Human renal cortical proximal tubular epithelial HK-2 cells (STCC10303P, Servicebio) were maintained in a humidified incubator (Thermo Fisher; MA, USA) at 37 °C with 5% CO 2 , and cultured in DMEM/F-12 medium (G4612) supplemented with 10% fetal bovine serum (G8003) and 1% penicillin-streptomycin (G4003).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: Elucidating the therapeutic efficacy and mechanisms of arctigenin in ameliorating renal fibrosis: a combined transcriptomic and proteomic study

doi: 10.3389/fphar.2026.1796732

Figure Lengend Snippet: ATG ameliorates UUO-induced RF and TGF-β1-induced fibrosis in HK-2 cells by regulating the S100A8/A9/NOX/NF-κB signaling pathway, which modulates TCA cycle and oxidative phosphorylation disruptions, thereby suppressing inflammation and oxidative stress-driven EMT. (A–C) Expression levels of TCA cycle-related factors (CA, NAD-MDH, NAD-ME) in renal tissues. (D–F) Expression levels of oxidative phosphorylation-related factors (ATP, CK, NADK) in renal tissues. (G–I) Relative mRNA expression levels of inflammatory factors (TNF-α, IL-6, IL-1β) in renal tissues detected by RT-qPCR. (J–L) Expression levels of oxidative stress markers (MDA, SOD, GSH) in renal tissues. (M–O) Relative mRNA expression levels of EMT markers (E-cadherin, N-cadherin, Vimentin) in renal tissues detected by RT-qPCR. (P–R) Expression levels of TCA cycle-related factors (CA, NAD-MDH, NAD-ME) in HK-2 cells. (S–U) Expression levels of oxidative phosphorylation-related factors (ATP, CK, NADK) in HK-2 cells. (V–X) Relative mRNA expression levels of inflammatory factors (TNF-α, IL-6, IL-1β) in HK-2 cells detected by RT-qPCR. (Y-AA) Relative levels of ROS in HK-2 cells measured by flow cytometry. (AB-AD) Relative mRNA expression levels of EMT markers (E-cadherin, N-cadherin, Vimentin) in HK-2 cells detected by RT-qPCR. Data are presented as mean ± SEM, n = 6 per group ( n = 3 per group for P-AD), * p < 0.05, ** p < 0.01, *** p < 0.001, ns, no significant.

Article Snippet: Human renal cortical proximal tubular epithelial HK-2 cells (STCC10303P, Servicebio) were maintained in a humidified incubator (Thermo Fisher; MA, USA) at 37 °C with 5% CO 2 , and cultured in DMEM/F-12 medium (G4612) supplemented with 10% fetal bovine serum (G8003) and 1% penicillin-streptomycin (G4003).

Techniques: Phospho-proteomics, Expressing, Quantitative RT-PCR, Flow Cytometry