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Related Experiment Videos

Oxysterol-induced endothelial cell dysfunction in culture.

S Ramasamy1, G A Boissonneault, B Hennig

  • 1Department of Nutrition and Food Science, University of Kentucky, Lexington 40506-0054.

Journal of the American College of Nutrition
|October 1, 1992
PubMed
Summary

Cholesterol oxidation products like Triol impair vascular endothelial barrier function. This study shows Triol alters membrane enzyme activity, specifically increasing Na+, K+, Mg(++)-ATPase and Ca(++)-ATPase, contributing to endothelial dysfunction.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Cardiovascular Research

Background:

  • Cholesterol oxidation products (oxysterols) are implicated in atherosclerosis.
  • Oxysterols, such as cholestan-3 beta,5 alpha,6 beta-triol (Triol), may promote atherosclerosis by disrupting vascular endothelial barrier function.
  • Endothelial dysfunction can arise from altered membrane characteristics and enzyme activities.

Purpose of the Study:

  • To investigate the impact of Triol on endothelial cell membrane-associated enzyme activities.
  • To determine the time- and concentration-dependent effects of Triol on endothelial barrier function and enzyme activity.

Main Methods:

  • Endothelial cell monolayers were incubated with varying concentrations of Triol.
  • Activities of membrane enzymes including Ca(++)-ATPase, Na+, K+, Mg(++)-ATPase, 5'-nucleotidase, and ACE were measured over 24 hours.

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  • Cellular toxicity was assessed by [3H]adenine release.

    • Main Results:

    • Triol exposure increased transendothelial albumin transfer, indicating decreased barrier function.
    • Activities of Ca(++)-ATPase and Na+, K+, Mg(++)-ATPase were significantly elevated after short incubation periods (2-4 hours).
    • 5'-nucleotidase activity increased after 24 hours, while ACE activity remained unchanged. Significant toxicity was observed at 24 hours.

    Conclusions:

    • Triol alters the activity of specific membrane-bound enzymes in endothelial cells.
    • Increased Ca(++)-ATPase and Na+, K+, Mg(++)-ATPase activities are early responses to Triol exposure.
    • These enzymatic alterations may contribute to the observed endothelial dysfunction and atherogenic potential of Triol.