nrk52e rat renal epithelial cells  (ATCC)

Journal: Journal of Translational Autoimmunity

Article Title: T-cell immunity in the experimental autoimmune vasculitis rat model

doi: 10.1016/j.jtauto.2025.100305

Figure Lengend Snippet: The anti-IL-17A antibody clone eBio17CK15A5 inhibits the biological activity of IL- 17A in vitro . (A,B) The rat renal tubular cell line (NRK52E cells) was stimulated with IL-17A or TNFα+IL-17A in absence or presence of neutralizing IL-17A antibody (n = 4 independent experiments). Unstimulated cells served as control. The gene expression of the IL-17A target genes CCL20 and CXCL1 served as readout. Stimulation with IL-17A induced the gene expression of CCL20 and CXCL1 already after 24 h. Co-stimulation with TNFα further increased CCL20 and CXCL1 transcripts. The presence of IL-17A antibody led in all cases to a reduction of CCL20 and CXCL1 transcripts confirming the inhibitory activity of the antibody.

Article Snippet: NRK52E cells (Leibniz Institute, DSMZ, Braunschweig, Germany) were cultured in complete DMEM supplemented with 5 % fetal calf serum (Biowest) and 1 % penicillin/streptomycin. (Thermo Fisher Scientific).

Techniques: Activity Assay, In Vitro, Control, Gene Expression

Journal: MedComm

Article Title: Benzoxazole Derivatives as Potent FXR and PPARα Dual Agonists With Anti‐Fibrotic and Metabolic Regulatory Effects

doi: 10.1002/mco2.70442

Figure Lengend Snippet: MHY5396 exerts anti‐inflammatory effects in renal epithelial cells and mitigates fibrotic responses in renal fibroblasts. (A) Effect of MHY5396 on FXR transcriptional activity was measured using the luciferase system in NRK52E cells ( n = 6). *** p < 0.001 compared to the pcDNA group. ## p < 0.005, ### p < 0.001 compared to the FXR promoter only group. (B) Effect of MHY5396 on PPARα transcriptional activity was measured using the PPRE luciferase system in NRK52E cells ( n = 5). *** p < 0.001 compared to the PPRE group. ### p < 0.001 compared to the PPRE+PPARα group. (C) Relative mRNA levels of chemokine genes ( Ccl2, Cxcl1 , and Il8 ) in lipopolysaccharide (LPS)‐treated NRK52E cells with or without MHY5396 (20 µM) treatment ( n = 5). ** p < 0.005, *** p < 0.001 compared to the control group. # p < 0.05, ## p < 0.005 compared to the LPS‐treated group. (D) NF‐κB transcriptional activity was assessed using the luciferase system in LPS‐treated NRK52E cells with or without MHY5396 (20 µM) treatment ( n = 6). ### p < 0.001 compared to the NF‐κB group. * p < 0.05 compared to the LPS‐treated group. (E) Protein levels of p65 and p‐p65 were determined in LPS‐treated NRK52E cells with or without MHY5396 (20 µM) treatment ( n = 3). GAPDH was used as the loading control. ** p < 0.001 compared to the control group. # p <0.05 compared to the LPS‐treated group. (F) Representative IF images of p65 expression (green) in LPS‐treated NRK52E cells with or without MHY5396 (20 µM) treatment. The nuclei were counterstained with DAPI. (G) Relative mRNA levels of fibrosis‐related genes ( Acta2, Col1a2 , and Col3a1 ) in TGFβ‐treated NRK49F cells with or without MHY5396 (20 µM) treatment ( n = 5). *** p < 0.001 compared to the control group. # p < 0.05, ## p < 0.005 compared to the TGFβ‐treated group. (H) Representative IF images of αSMA expression (green) in TGFβ‐treated NRK49F cells with or without MHY5396 (20 µM) treatment. (I) Protein levels of αSMA and vimentin in TGFβ‐treated NRK49F cells with or without MHY5396 (20 µM) treatment ( n = 3). GAPDH was used as the loading control. Relative protein expressions were quantified using densitometry. * p < 0.05, ** p < 0.005, *** p < 0.001 compared to the control group. # p < 0.05 compared to the TGFβ‐treated group.

Article Snippet: To assess the effect of MHY5396 on kidney cells, experiments were conducted using NRK52E cells, a rat kidney epithelial cell line, and NRK49F cells, a rat kidney fibroblast cell line, both obtained from ATCC.

Techniques: Activity Assay, Luciferase, Control, Expressing

Journal: Molecular Medicine Reports

Article Title: β-elemene attenuates IRI-AKI by inhibiting inflammation and apoptosis via suppression of the TLR4/MyD88/NF-κB/MAPK signal axis activation

doi: 10.3892/mmr.2025.13586

Figure Lengend Snippet: β-elemene protects NRK52E cells from H 2 O 2 -induced inflammation. (A) Optimal duration of H 2 O 2 treatment in NRK52E cells was identified by employing the CCK-8 assay. (B) CCK-8 analysis was employed to identify the ELE impact on viability of NRK52E cells. (C) NRK52E cells underwent pretreatment with various doses of ELE and were treated for 6 h with or without 600 µM H 2 O 2 . CCK-8 analysis was used to determine cell viability. Reverse transcription-quantitative PCR was employed to evaluate the mRNA expression of (D) IL-1β, (E) TNF-α, (F) MCP-1 and (G) ICAM-1. *P<0.05, **P<0.01, ***P<0.001. CCK-8, Cell Counting Kit-8; OD, optical density; ELE, β-elemene; ns, not significant; MCP-1, monocyte chemoattractant protein-1; ICAM-1, intercellular adhesion molecule 1.

Article Snippet: Rat proximal tubular epithelial NRK52E cells were obtained from American Type Culture Collection.

Techniques: CCK-8 Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Cell Counting

Journal: Molecular Medicine Reports

Article Title: β-elemene attenuates IRI-AKI by inhibiting inflammation and apoptosis via suppression of the TLR4/MyD88/NF-κB/MAPK signal axis activation

doi: 10.3892/mmr.2025.13586

Figure Lengend Snippet: ELE inhibits the inflammatory response by suppressing TLR4/MyD88/NF-κB pathway activation in vivo and in vitro . (A) Renal p-P65, MyD88, TLR4 and P65 expression. (B) Western blotting of the p-P65, MyD88, TLR4 and P65 expression in NRK52E cells treated with H 2 O 2 and various concentrations of ELE. *P<0.05, **P<0.01. ELE, β-elemene; IRI, ischemia-reperfusion injury; TLR4, toll-like receptor 4; MyD88, myeloid differentiation primary response gene 88; p-, phosphorylated-; ns, not significant.

Article Snippet: Rat proximal tubular epithelial NRK52E cells were obtained from American Type Culture Collection.

Techniques: Activation Assay, In Vivo, In Vitro, Expressing, Western Blot

Journal: Molecular Medicine Reports

Article Title: β-elemene attenuates IRI-AKI by inhibiting inflammation and apoptosis via suppression of the TLR4/MyD88/NF-κB/MAPK signal axis activation

doi: 10.3892/mmr.2025.13586

Figure Lengend Snippet: ELE ameliorates H 2 O 2 -induced NRK52E cell apoptosis. (A) TUNEL assay; scale bar, 50 µm. (B) Bax, c-caspase3, Bcl-2 and caspase-3 protein expression were measured using western blotting. (C) CCK-8 analysis was performed to identify the NAC impact on viability of NRK52E cells. (D) Identification and semi-quantification of Bax, c-caspase3, Bcl-2 and caspase-3 protein expression in NRK52E cells. *P<0.05, **P<0.01. ELE, β-elemene; c-, cleaved; ns, not significant; OD, optical density; NAC, N-Acetylcysteine.

Article Snippet: Rat proximal tubular epithelial NRK52E cells were obtained from American Type Culture Collection.

Techniques: TUNEL Assay, Expressing, Western Blot, CCK-8 Assay

Journal: Molecular Medicine Reports

Article Title: β-elemene attenuates IRI-AKI by inhibiting inflammation and apoptosis via suppression of the TLR4/MyD88/NF-κB/MAPK signal axis activation

doi: 10.3892/mmr.2025.13586

Figure Lengend Snippet: ELE decreases inflammatory and apoptosis signaling by inhibiting MAPK signaling pathway activation. (A) ERK, p-JNK, p-P38, JNK, p-ERK and P38 in the kidney. (B) Identification and semi-quantification of p-JNK, p-ERK1/2, p-P38, JNK, ERK1/2 and P38 expression in NRK52E cells. *P<0.05, **P<0.01. ELE, β-elemene; IRI, ischemia-reperfusion injury; ns, not significant; p-, phosphorylated.

Article Snippet: Rat proximal tubular epithelial NRK52E cells were obtained from American Type Culture Collection.

Techniques: Activation Assay, Expressing

Journal: Molecular Medicine Reports

Article Title: β-elemene attenuates IRI-AKI by inhibiting inflammation and apoptosis via suppression of the TLR4/MyD88/NF-κB/MAPK signal axis activation

doi: 10.3892/mmr.2025.13586

Figure Lengend Snippet: MyD88 knockdown suppresses apoptosis and MAPK signaling pathway activation in H 2 O 2 -treated NRK52E cells. (A) MyD88 expression in NRK52E cells. (B) MyD88, Bax, c-caspase3, Bcl-2, JNK, caspase-3 p-ERK, p-JNK, ERK, p-P38 and P38 expression in NRK52E cells treated with H 2 O 2 . *P<0.05, **P<0.01. c-, cleaved; p-, phosphorylated; ns, not significant; si, small interfering.

Article Snippet: Rat proximal tubular epithelial NRK52E cells were obtained from American Type Culture Collection.

Techniques: Knockdown, Activation Assay, Expressing

Journal: iScience

Article Title: Injured tubular epithelia-derived CCN1 promotes the mobilization of fibroblasts toward injury sites after kidney injury

doi: 10.1016/j.isci.2025.112176

Figure Lengend Snippet: PT-specific transcriptomics reveal Ccn1 up-regulation in the early phase after cisplatin-induced kidney injury in vivo (A) Experimental scheme. Bigenic mice (SLC34a1GCE × R26tdTomato) received intraperitoneal injections of cisplatin (15 mg/kg) and tamoxifen (3 mg/kg at each time point) as indicated. (B) Changes in BUN after the administration of cisplatin. n = 4–5 per group. (C) Isolation of tdTomato+ tubular epithelial cells using FACS. (D) Histological analysis of the kidney after the cisplatin injection. PAS staining of kidney sections and immunostaining of LTL and KIM1. The scale bars indicate 50 μm in PAS staining, 100 μm in low-power field pictures and 20 μm in high-power field pictures in immunostaining. (E) Time-course analyses of qPCR of RNA from the whole kidney for the representative markers of tubular injury ( Havcr1 ), profibrotic factors ( Ccn1 , Tgfb1 , Ctgf , and Pdgfb ), myofibroblast ( Acta2 ), mesenchymal cell ( Vim ), and extracellular matrix ( Col1a1 and Fn1 ). n = 4–5 per group. (F) Time-course analyses of qPCR of RNA from the isolated tdTomato+ tubular epithelial cells. n = 4–5 per group. For all groups, data are means ± SEM. Statistical analyses were performed by unpaired t test for comparisons to day 0 (B, E, F). ∗ p < 0.05 vs. day 0.

Article Snippet: Rat kidney epithelial cells (NRK52E cells) , The JCRB Cell Bank , N/A.

Techniques: In Vivo, Isolation, Injection, Staining, Immunostaining

Journal: iScience

Article Title: Injured tubular epithelia-derived CCN1 promotes the mobilization of fibroblasts toward injury sites after kidney injury

doi: 10.1016/j.isci.2025.112176

Figure Lengend Snippet: The PT-specific knockout of CCN1 inhibits the accumulation of fibroblasts at injured tubules in cisplatin nephrotoxicity in vivo (A) Experimental scheme. Tubular-specific CCN1 knockout mice (CCN1Flox/Flox SLC34a1GCE x R26tdTomato: CCN-KO) received intraperitoneal injections of cisplatin (15 mg/kg) and tamoxifen (3 mg/kg at each time point) as indicated. (B) Histological analysis of the kidney after the cisplatin injection by immunostaining of KIM1. (C) RT-PCR of the Ccn1 and Havcr1 genes in sorted tdTomato+ tubular epithelial cells. (D) Changes in BUN after the administration of cisplatin. n = 7–8 per group. (E) Representative co-immunofluorescence images of KIM1 and pFAK, PDGFRβ and pFAK, and PDGFRβ and γH2AX at 4 days after cisplatin injection. (F) Representative images of Sirius red staining at 14 days after cisplatin injection. (G) Semi-quantitative fibrosis scores. n = 9 WT Cis (−), n = 8 WT Cis (+), n = 7 CCN1-KO Cis (+). (H) qPCR of RNA from whole kidneys as representative markers of myofibroblasts ( Acta2 ), fibroblasts ( Pdgfrb ), profibrotic factor ( Tgfb1 ), macrophage ( Cd68 ), tubular injury ( Havcr1 ), tubular integrity ( Lrp2 ), and extracellular matrix ( Col1a1 and Fn1 ). n = 5 WT Cis (−), n = 8 WT Cis (+), n = 7 CCN1-KO Cis (+). The scale bars indicate 100 μm in (B), 50 μm in low-power field pictures, and 20 μm in high-power field pictures in (E). The scale bars indicate 50 μm in (F). For all groups, data are means ± SEM. Statistical analyses were performed by unpaired t test for comparisons of two variables and by ANOVA and Dunnett’s post hoc test for comparisons of multiple variables. ∗ p < 0.05.

Article Snippet: Rat kidney epithelial cells (NRK52E cells) , The JCRB Cell Bank , N/A.

Techniques: Knock-Out, In Vivo, Injection, Immunostaining, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Staining

Journal: iScience

Article Title: Injured tubular epithelia-derived CCN1 promotes the mobilization of fibroblasts toward injury sites after kidney injury

doi: 10.1016/j.isci.2025.112176

Figure Lengend Snippet: The PT-specific knockout of CCN1 inhibits the accumulation of fibroblasts at injured tubules and subsequent tissue fibrosis in IRI in vivo (A) Experimental scheme. Tubular-specific CCN1 knockout mice (CCN1Flox/Flox SLC34a1GCE x R26tdTomato: CCN-KO) received IRI (35 min) and tamoxifen (3 mg/kg at each time point) as indicated. (B) Histological analysis of the kidney at 1 day after IRI by immunostaining of KIM1. (C) RT-PCR of the Ccn1 and Havcr1 genes in sorted tdTomato+ tubular epithelial cells. (D) Representative co-immunofluorescence images of KIM1 and pFAK, and PDGFRβ and pFAK at 1 day after IRI. (E) Representative images of Sirius red staining. (F) Semi-quantitative fibrosis scores at 14 days after IRI. n = 9 WT IRI (−), n = 8 WT IRI (+), n = 9 CCN1-KO IRI (−), n = 8 CCN1-KO IRI (+). (G) qPCR of RNA from whole kidneys as representative markers of tubular injury ( Havcr1 ), tubular integrity ( Lrp2 ), chemokine ( Ccl2 ), macrophage ( Cd68 ), fibroblasts ( Pdgfrb ), myofibroblasts ( Acta2 ), extracellular matrix ( Col1a1 and Fn1 ), and profibrotic factor ( Tgfb1 ). n = 9 WT IRI (−), n = 8 WT IRI (+), n = 8 CCN1-KO IRI (−), n = 7 CCN1-KO IRI (+). The scale bars indicate 100 μm in (B), 50 μm in low-power field pictures, and 20 μm in high-power field pictures in (D). The scale bars indicate 50 μm in (E). For all groups, data are means ± SEM. Statistical analyses were performed by paired t test for comparisons of IRI kidney and contralateral kidney (CLK) and by ANOVA and Dunnett’s post hoc test for comparisons of multiple variables. ∗ p < 0.05.

Article Snippet: Rat kidney epithelial cells (NRK52E cells) , The JCRB Cell Bank , N/A.

Techniques: Knock-Out, In Vivo, Immunostaining, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Staining

Journal: iScience

Article Title: Injured tubular epithelia-derived CCN1 promotes the mobilization of fibroblasts toward injury sites after kidney injury

doi: 10.1016/j.isci.2025.112176

Figure Lengend Snippet:

Article Snippet: Rat kidney epithelial cells (NRK52E cells) , The JCRB Cell Bank , N/A.

Techniques: Plasmid Preparation, Virus, CRISPR, Recombinant, Inhibition, Negative Control, Saline, Western Blot, Proliferation Assay, Sample Prep, Software, Microscopy