Scavengers of reactive oxygen species, paracalcitol, RhoA, and Rac-1 inhibitors and tacrolimus inhibit angiotensin II-induced actions on glomerular permeability

Am J Physiol Renal Physiol. 2013 Aug 1;305(3):F237-43. doi: 10.1152/ajprenal.00154.2013. Epub 2013 May 8.

Abstract

Systemic infusions of ANG II rapidly induce large, dynamic increases in the permeability of the glomerular filtration barrier (GFB) in rats. After binding to its receptor(s), ANG II generates reactive oxygen species (ROS) and produces Ca²⁺ influx into cells, leading to activation of a plethora of signaling cascades, including, e.g., calcineurin and small GTPases, such as Rac-1 and RhoA. In the present study we sought to interact with some of these cascades to test potential novel antiproteinuric agents. In anesthetized Wistar rats, the left urether was cannulated for urine collection, and blood access was achieved. Rats were infused with ANG II (16 ng·kg⁻¹·min⁻¹) alone, or together with the ROS scavengers tempol or dimethylthiourea (DMTU) or the D-vitamin analog paracalcitol, the RhoA-kinase inhibitor Y-27632, the Rac-1 inhibitor NSC-23766, or the calcineurin inhibitor tacrolimus. FITC-Ficoll-70/400 (mol.radius 10-80 Å) and ⁵¹Cr-EDTA were infused throughout the experiment. Plasma and urine samples were taken during baseline and at 5 and 15 min after the start of the infusions and analyzed by high-performance size-exclusion chromatography for determination of glomerular sieving coefficients (θ) for Ficoll10-80Å. ANG II infusion into rats caused marked increases in glomerular permeability to large Ficoll molecules (Ficoll50-80Å), which were abrogated by the ROS scavenger tempol and partly by DMTU. Paracalcitol, RhoA, and Rac-1 inhibition, and, to some extent tacrolimus, but not prostacyclin, could also inhibit the glomerular permeability actions of ANG II. Our data suggest that cellular ROS generation and active Ca²⁺ signaling are involved in ANG II-induced increases in glomerular permeability.

Keywords: Ficoll; capillary permeability; glomerular sieving coefficient; microalbuminuria; two-pore model.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiotensin II / physiology*
  • Angiotensin Receptor Antagonists*
  • Animals
  • Calcineurin Inhibitors
  • Chromatography, Gel
  • Cyclic N-Oxides / pharmacology
  • Epoprostenol / pharmacology
  • Ergocalciferols / pharmacology*
  • Fluorescein-5-isothiocyanate
  • Free Radical Scavengers / pharmacology*
  • GTP Phosphohydrolases / antagonists & inhibitors
  • GTP Phosphohydrolases / metabolism
  • Glomerular Filtration Rate / drug effects
  • Immunosuppressive Agents / pharmacology*
  • Kidney Glomerulus / drug effects*
  • Kidney Glomerulus / metabolism*
  • Male
  • Permeability / drug effects
  • Rats
  • Rats, Wistar
  • Reactive Oxygen Species / metabolism*
  • Spin Labels
  • Tacrolimus / pharmacology*
  • Thiourea / analogs & derivatives
  • Thiourea / pharmacology
  • rac1 GTP-Binding Protein / antagonists & inhibitors*
  • rho-Associated Kinases / antagonists & inhibitors*
  • rho-Associated Kinases / metabolism*

Substances

  • Angiotensin Receptor Antagonists
  • Calcineurin Inhibitors
  • Cyclic N-Oxides
  • Ergocalciferols
  • Free Radical Scavengers
  • Immunosuppressive Agents
  • Reactive Oxygen Species
  • Spin Labels
  • Angiotensin II
  • paricalcitol
  • 1,3-dimethylthiourea
  • Epoprostenol
  • rho-Associated Kinases
  • GTP Phosphohydrolases
  • Rac1 protein, rat
  • rac1 GTP-Binding Protein
  • Thiourea
  • Fluorescein-5-isothiocyanate
  • tempol
  • Tacrolimus