Overexpression of PHGPx inhibits hydroperoxide-induced oxidation, NFkappaB activation and apoptosis and affects oxLDL-mediated proliferation of rabbit aortic smooth muscle cells

Atherosclerosis. 2000 Oct;152(2):307-16. doi: 10.1016/s0021-9150(99)00486-4.

Abstract

Rabbit abdominal aortic smooth muscle cells (SMC) were stably transfected with the cDNA of porcine phospholipid hydroperoxide glutathione peroxidase (PHGPx) by means of a retroviral gene transfer technique, to create a model for studying cellular processes relevant to atherogenesis. The transfected cells (SMC/PHGPx) had approximately 4-fold higher PHGPx activity when cultured in the presence of selenite whereas the parental cells did not show any significant increase in PHGPx or total GPx activity upon selenium supplementation. In situ functionality of PHGPx was validated by inhibition of linoleic acid hydroperoxide-induced toxicity, dihydrorhodamine oxidation, NFkappaB activation and apoptosis. SMC grown in 1% FCS responded to oxidized LDL (oxLDL) with a marked proliferation, as measured by [3H]thymidine incorporation, irrespective of selenium supplementation. In SMC/PHGPx grown with or without selenite under control conditions or exposed to native LDL, thymidine incorporation was generally depressed. Also, oxLDL-induced proliferation was lower in SMC/PHGPx compared to untransfected SMC up to 24 h of incubation. After 40 h, however, selenite supplementation restored maximum proliferation response to oxLDL in SMC/PHGPx. The results suggest a proliferative effect of endogenous hydroperoxides in SMC. They further reveal that hydroperoxy lipids of oxLDL contribute to the induction of proliferation, but also suggest involvement of hydroxy lipids in the response to oxLDL.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aorta, Abdominal / cytology*
  • Aorta, Abdominal / enzymology*
  • Apoptosis / drug effects
  • Cell Division / drug effects
  • Cells, Cultured
  • Gene Transfer Techniques
  • Glutathione Peroxidase / biosynthesis*
  • Glutathione Peroxidase / metabolism
  • Hydrogen Peroxide / pharmacology
  • Lipoproteins, LDL / pharmacology*
  • Muscle, Smooth, Vascular / cytology*
  • Muscle, Smooth, Vascular / enzymology*
  • NF-kappa B / metabolism*
  • Oxidants / pharmacology
  • Oxidation-Reduction
  • Phospholipid Hydroperoxide Glutathione Peroxidase
  • Rabbits
  • Selenium / pharmacology

Substances

  • Lipoproteins, LDL
  • NF-kappa B
  • Oxidants
  • oxidized low density lipoprotein
  • Hydrogen Peroxide
  • Phospholipid Hydroperoxide Glutathione Peroxidase
  • Glutathione Peroxidase
  • Selenium