Journal: Journal of Diabetes Research

Article Title: Sweet Taste Receptors Mediated ROS-NLRP3 Inflammasome Signaling Activation: Implications for Diabetic Nephropathy

doi: 10.1155/2018/7078214

Figure Lengend Snippet: High glucose inhibited the expression of STRs and activated NLRP3 inflammasome. (a) GMCs (SV-40 MES 13) and human proximal tubular cells (HK-2) were treated with 30 mmol/L high glucose for 6, 12, and 24 h, and Western blot or qRT-PCR was performed to detect T1R2 and T1R3 protein or mRNA expression levels. (b) Cells were treated with the indicated concentrations of glucose or mannitol for 24 h. Western blot or qRT-PCR was performed to detect T1R2 and T1R3 protein or mRNA expression levels. (c) Western blotting analysis of NLRP3, cleaved caspase-1, and cleaved IL-1 β in GMCs treated with an indicated concentration of glucose or mannitol for 24 h. (d) IL-1 β protein level in the GMC culture supernatant was determined by ELISA. Data are expressed as mean ± SD. ∗ p < 0.05 versus NC group.

Article Snippet: Mouse GMCs (SV40 MES 13) and human proximal tubular cells (HK-2) were purchased from the China Center for Type Culture Collection (CCTCC) and were cultured in Dulbecco's modified Eagle's medium (DMEM, Glbco) containing 5.6 mmol/L glucose and 10% fetal bovine serum (FBS, Glbco) at 37°C and 5% CO 2.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Diabetes Research

Article Title: Sweet Taste Receptors Mediated ROS-NLRP3 Inflammasome Signaling Activation: Implications for Diabetic Nephropathy

doi: 10.1155/2018/7078214

Figure Lengend Snippet: High glucose-induced downregulation of STRs and associated signaling components was reversed by lactisole. Mouse GMCs (SV-40 MES 13) was firstly incubated with 3, 5, and 10 mM lactisole; then 30 mM high glucose (HG) alone was intervened by the indicated concentrations of lactisole following 30 mM high glucose for 24 h. (a) Cell viability was determined by MTT analysis. (b) The protein and mRNA expression of T1R2 and T1R3 in GMCs were detected by Western blotting and qRT-PCR. (c) The protein expression of T1R2 and T1R3 in human proximal tubular cells (HK-2) was detected by Western blot. (d) STR-associated signaling components ( α -gustducin, PLC β 2, and TRPM5) were detected by qRT-PCR. (e) GMCs were stimulated with 30 mmol/L glucose in the presence (blue), and absence (red) of 5 mmol/L lactisole, and the Ca 2+ influx changes in Ca 2+ dye fluorescence was monitored. Data are expressed as mean ± SD. ∗ p < 0.05 versus NC group. # p < 0.05 versus 30 mM glucose (HG) stimulation group.

Article Snippet: Mouse GMCs (SV40 MES 13) and human proximal tubular cells (HK-2) were purchased from the China Center for Type Culture Collection (CCTCC) and were cultured in Dulbecco's modified Eagle's medium (DMEM, Glbco) containing 5.6 mmol/L glucose and 10% fetal bovine serum (FBS, Glbco) at 37°C and 5% CO 2.

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

Journal: Oncotarget

Article Title: A novel pan-Nox inhibitor, APX-115, protects kidney injury in streptozotocin-induced diabetic mice: possible role of peroxisomal and mitochondrial biogenesis

doi: 10.18632/oncotarget.18540

Figure Lengend Snippet: Diabetes was induced in mice by intraperitoneal injection of STZ (50 mg/kg). Then APX-115 (60 mg/kg/day) or losartan (1.5 mg/kg/day) was administered orally for 12 weeks to diabetic mice. After 12 weeks, urine and blood samples were collected for analysis of ( A ) urinary albumin excretion, ( B ) albumin/creatinine ratio, ( C ) creatinine clearance rate, and ( D ) plasma cystatin C. ( E ) Kidneys were fixed in paraffin and cut into 3 μm sections that were subsequently stained with PAS reagent. Scale bar: 10 μm; original magnification: 630×. After PAS staining, ( F ) glomerular volume, ( G ) mesangial area, and ( H ) tuft area were analyzed using Image-Pro Plus 4.5.1. DM, STZ-induced diabetic mice. Data are presented as means ± SE of 10–12 mice/group; * p < 0.05 vs. control, † p < 0.05 vs. DM.

Article Snippet: A murine mesangial cell line (MES-13, cloned from mice transgenic for the early region of SV-40 virus, passage 25) was obtained from American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Injection, Clinical Proteomics, Staining, Control

Journal: Oncotarget

Article Title: A novel pan-Nox inhibitor, APX-115, protects kidney injury in streptozotocin-induced diabetic mice: possible role of peroxisomal and mitochondrial biogenesis

doi: 10.18632/oncotarget.18540

Figure Lengend Snippet: ( A ) Plasma LPO, ( B ) urinary LPO, ( C ) kidney tissue LPO, ( D ) Nox1, ( E ) Nox2, and ( F ) Nox4 mRNA expression levels in kidneys were measured using real-time PCR. ( G and H ) Frozen kidney sections were stained with DHE at 5 µM (original magnification: 400×; scale bar: 20 μm). (A–H) Data are presented as means ± SE of 10–12 mice/group; * p < 0.05 vs. control, † p < 0.05 vs. DM. ( I ) Mesangial cells were incubated with or without APX-115 (1 µM) for 30 min and stimulated with or without 30 mM high glucose (HG) for 24 h followed by angII for 30 min. After that cells were incubated with 10 µM DCF-DA for 10 min and the fluorescence intensity was measured with a Zeiss vision system. Data are presented as means ± SE of at least 2 independent experiments; * p < 0.05 vs. control, † p < 0.05 vs. angII or angII+HG in DMSO.

Article Snippet: A murine mesangial cell line (MES-13, cloned from mice transgenic for the early region of SV-40 virus, passage 25) was obtained from American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Clinical Proteomics, Expressing, Real-time Polymerase Chain Reaction, Staining, Control, Incubation, Fluorescence