title: High glucose modifies transient receptor potential canonical type 6 channels via increased oxidative stress and syndecan-4 in human podocytes.
abstract: Transient receptor potential canonical (TRPC) channels type 6 play an important role in the function of human podocytes. Diabetic nephropathy is characterized by altered TRPC6 expression and functions of podocytes. Thus, we hypothesized that high glucose modifies TRPC6 channels via increased oxidative stress and syndecan-4 (SDC-4) in human podocytes. Human podocytes were exposed to control conditions (5.6 mmol/L D-glucose), high glucose (30 mmol/L D-glucose or L-glucose), 100 μmol/L peroxynitrite, or high glucose and the superoxide dismutase mimetic tempol (100 μmol/L). TRPC6 and SDC-4 transcripts and protein expression were measured using RT-PCR and in-cell Western assay. Intracellular reactive oxygen species (ROS) and cytosolic calcium were measured using fluorescent dye techniques. High D-glucose increased TRPC6 transcripts to 8.66±4.08 (p<0.05) and TRPC6 protein expression to 1.44±0.07 (p<0.05) without altering SDC-4 transcripts or protein expression. The D-glucose induced increase of TRPC6 expression was blocked by tempol. Increased oxidative stress using peroxynitrite significantly increased TRPC6 transcripts to 4.29±1.26 (p<0.05) and TRPC6 protein expression to 1.28±0.05 (p<0.05) without altering SDC-4 transcripts or protein expression. In human podocytes transfected with scrambled siRNA, high D-glucose increased ROS after 90 min to 3.55±0.08 arbitrary units while 5.6 mmol/L D-glucose increased ROS to 2.49±0.09 (p<0.001) only. The increase in ROS was inhibited by tempol and by SDC-4 knockdown. High glucose modifies TRPC6 channels and ROS production via SDC-4 in human podocytes.
authors.affiliation: Experimental and Clinical Research Center, Charité, Berlin, Germany; Charité Berlin, Shear Stress and Vascular Biology Research Group, Institute of Physiology, Berlin, Germany. Electronic address: Florian.Thilo@charite.de.
authors.forename: Florian
authors.initials: F
authors.lastname: Thilo
authors.affiliation: Experimental and Clinical Research Center, Charité, Berlin, Germany; Charité Berlin, Shear Stress and Vascular Biology Research Group, Institute of Physiology, Berlin, Germany.
authors.forename: Marlene
authors.initials: M
authors.lastname: Lee
authors.affiliation: Odense University Hospital, Department of Nephrology, University of Southern Denmark, Institute for Molecular Medicine, Cardiovascular and Renal Research, Institute of Clinical Research, Odense, Denmark; Department of Urology, Tenth People's Hospital, Tongji University of Shanghai, PR China.
authors.forename: Shengqiang
authors.initials: S
authors.lastname: Xia
authors.affiliation: Charité Berlin, Shear Stress and Vascular Biology Research Group, Institute of Physiology, Berlin, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Zakrzewicz
authors.affiliation: Odense University Hospital, Department of Nephrology, University of Southern Denmark, Institute for Molecular Medicine, Cardiovascular and Renal Research, Institute of Clinical Research, Odense, Denmark.
authors.forename: Martin
authors.initials: M
authors.lastname: Tepel
mesh_terms: D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005947:Glucose / Q000008:administration & dosage*; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology*; D010100:Oxygen / Q000378:metabolism*; D050199:Podocytes / Q000187:drug effects / Q000378:metabolism*; D053671:Syndecan-4 / Q000378:metabolism*; D050052:TRPC Cation Channels / Q000187:drug effects / Q000378:metabolism*; D000074287:TRPC6 Cation Channel
chemical_list: C508304:SDC4 protein, human; D053671:Syndecan-4; D050052:TRPC Cation Channels; D000074287:TRPC6 Cation Channel; C117032:TRPC6 protein, human; D005947:Glucose; D010100:Oxygen
keywords: Diabetic nephropathy; High glucose; Oxidative stress; Podocytes; Syndecan-4; Transient receptor potential channels