Inhibition of apoptotic signalling in spermine-treated vascular smooth muscle cells by a novel glutathione precursor

Cell Biol Int. 2010 Apr 1;34(5):503-11. doi: 10.1042/CBI20090349.

Abstract

CKD (chronic kidney disease) is a public health problem, mediated by haemodynamic and non-haemodynamic events including oxidative stress. We investigated the effect of two GSH (glutathione) precursors, NAC (N-acetylcysteine) and cystine as the physiological carrier of cysteine in GSH with added selenomethionine (F1) in preventing spermine (uraemic toxin)-induced apoptosis in cultured human aortic VSMC (vascular smooth muscle cells). VSMCs exposed to spermine (15 microM) with or without antioxidants (doses 50, 100, 200 and 500 microg/ml) were assessed for apoptosis, JNK (c-Jun-NH2-terminal kinase) activation and iNOS (inducible nitric oxide synthase) induction and activation of intrinsic pathway signalling. Spermine exposure resulted in activation of JNK and iNOS induction and apoptosis. NAC and F1 (dose range 50-500 microg/ml) attenuated spermine-induced acceleration of VSMC apoptosis but only F1 (at 200 and 500 microg/ml) maintained spermine-induced apoptosis at control levels. Spermine-induced JNK activation was prevented by 200 microg/ml of both NAC and F1, while iNOS induction was blocked only by F1. Notably, the adverse effects of spermine on BAX/BCL-2 ratio, cytochrome c release and caspase activation was fully attenuated by F1. In conclusion, F1 was more effective than NAC in preventing spermine-induced apoptosis and downstream changes in related signal transduction pathways in VSMCs. Further studies are needed to examine the effect of these compounds in preventing CKD-associated vascular disease.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Acetylcysteine / metabolism
  • Acetylcysteine / pharmacology*
  • Antioxidants / metabolism
  • Antioxidants / pharmacology
  • Apoptosis / drug effects*
  • Cystine / metabolism
  • Cystine / pharmacology*
  • Enzyme Activation
  • Glutathione / analogs & derivatives*
  • Glutathione / metabolism
  • Humans
  • In Situ Nick-End Labeling
  • JNK Mitogen-Activated Protein Kinases / metabolism
  • Muscle, Smooth, Vascular / cytology
  • Myocytes, Smooth Muscle / cytology
  • Myocytes, Smooth Muscle / drug effects*
  • Myocytes, Smooth Muscle / physiology
  • Nitric Oxide Synthase Type II / metabolism
  • Oxidative Stress
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Renal Insufficiency, Chronic / complications
  • Renal Insufficiency, Chronic / metabolism
  • Renal Insufficiency, Chronic / pathology
  • Signal Transduction / drug effects*
  • Spermine / pharmacology*
  • bcl-2-Associated X Protein / metabolism

Substances

  • Antioxidants
  • Proto-Oncogene Proteins c-bcl-2
  • bcl-2-Associated X Protein
  • Spermine
  • Cystine
  • Nitric Oxide Synthase Type II
  • JNK Mitogen-Activated Protein Kinases
  • Glutathione
  • Acetylcysteine