Puromycin aminonucleoside induces glomerular epithelial cell apoptosis

Exp Mol Pathol. 2001 Feb;70(1):54-64. doi: 10.1006/exmp.2000.2345.

Abstract

Glomerular epithelial cell (GEC) injury has been considered to play an important role in puromycin aminonucleoside (PAN)-induced nephrosis. We studied the effect of PAN on rat as well as human GEC apoptosis. Morphogic evaluation of GEC apoptosis and necrosis was carried out by staining with H-33342 and propidium iodide. GEC apoptosis was further confirmed by DNA fragmentation assay (by both agarose gel electrophoresis and end-labeling). To determine the dose- and time-response effect of PAN, GECs were treated with variable concentrations of PAN (10 to 500 microg/ml) for variable time periods (6 to 48 h). To determine the role of gene synthesis, we studied the effect of actinomycin D (a transcriptional inhibitor) on PAN-induced GEC apoptosis. To determine the role of free radicals, we evaluated the effect of superoxide dismutase (SOD), dimethylthiourea (DMTU), and catalase on PAN-induced GEC apoptosis. PAN induced GEC apoptosis in a dose- and time-dependent manner. PAN at a high concentration (PAN, 100 microg/ml) also induced a moderate degree of GEC necrosis. In DNA fragmentation assays PAN-treated GECs showed the classic ladder pattern. PAN-induced GEC apoptosis was partly attenuated with free radical scavengers, such as SOD, DMTU, and catalase. In addition, actinomycin D attenuated PAN-induced GEC apoptosis. PAN induces GEC apoptosis, which may be mediated through the generation of reactive oxygen species.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Catalase / pharmacology
  • Cells, Cultured
  • DNA / drug effects
  • DNA Fragmentation / drug effects
  • Dactinomycin / pharmacology
  • Disease Models, Animal
  • Dose-Response Relationship, Drug
  • Electrophoresis, Agar Gel
  • Epithelial Cells / drug effects
  • Epithelial Cells / pathology
  • Free Radical Scavengers / pharmacology
  • Humans
  • In Situ Nick-End Labeling
  • Kidney Glomerulus / drug effects*
  • Kidney Glomerulus / pathology
  • Puromycin Aminonucleoside / toxicity*
  • Rats
  • Rats, Sprague-Dawley
  • Superoxide Dismutase / pharmacology
  • Thiourea / analogs & derivatives*
  • Thiourea / pharmacology

Substances

  • Free Radical Scavengers
  • Dactinomycin
  • Puromycin Aminonucleoside
  • 1,3-dimethylthiourea
  • DNA
  • Catalase
  • Superoxide Dismutase
  • Thiourea