A comparison of the effects of monotherapy with rosuvastatin, atorvastatin or ezetimibe versus combination treatment with rosuvastatin-ezetimibe and atorvastatin-ezetimibe on the integrity of vascular endothelial cells damaged by oxidized cholesterol

PLoS One. 2021 Sep 7;16(9):e0256996. doi: 10.1371/journal.pone.0256996. eCollection 2021.

Abstract

Dyslipidemia, atherosclerosis, and cardiovascular events can be prevented, or treated, using statins, alone or in combination with ezetimibe. The aim of the study was to compare the direct pleiotropic effects of two commonly-used statins (atorvastatin, rosuvastatin), ezetimibe and their combinations on endothelial cells damaged by oxidized cholesterol. HUVEC cultures were stimulated for 20 hours with atorvastatin (5 μM; 2793 ng/mL), rosuvastatin (10 μM; 4815 ng/mL), ezetimibe (1.22 μM; 500 ng/mL), atorvastatin plus ezetimibe (5 μM + 1.22 μM; 2793 ng/mL + 500 ng/mL) and rosuvastatin plus ezetimibe (10 μM + 1.22 μM; 4815 ng/mL + 500ng/mL) in separate groups, with or without 25-hydroxycholesterol pre-incubation (24.83 μM; 10 μg/mL; four hours then washout). HUVEC integrity was measured in the RTCA-DP xCELLigence system. The mRNA expression and protein levels of ZO-1, OCLN, ICAM-1 were analyzed by real-time PCR and ELISA. Pre-incubation with 25-OHC resulted in decreased endothelial cell integrity (p<0.001), decreased expression of ZO-1 mRNA (p<0.05) and protein levels (p<0.05), OCLN mRNA (p<0.05) and protein levels (p<0.05) and increased ICAM-1 mRNA (p<0.001) and protein levels (p<0.001) compared to the control group. Incubation with rosuvastatin (12h p<0.01; 24h p<0.001) and atorvastatin (only 12h p<0.05) restored HUVEC integrity. Subsequent incubation with rosuvastatin increased ZO-1 mRNA (p<0.001) and protein (p<0.001) levels. Subsequent addition of ezetimibe increased ZO-1 mRNA level (p<0.001) but not protein level. Furthermore, only incubation with rosuvastatin increased OCLN mRNA (p<0.05) and protein (p<0.05) levels. In each drug-stimulated group, both ICAM-1 mRNA and protein levels were reduced after initial incubation with oxysterol (p<0.05). 25-hydroxycholesterol disrupts endothelial integrity, decreases the mRNA and protein levels of tight junction, and increases those of intercellular adhesion molecules. Both rosuvastatin and atorvastatin can improve endothelial integrity, but only rosuvastatin can completely abolish the effect of oxysterol. The combination of statins with ezetimibe has less direct effect on the endothelial barrier than the statins alone.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Atherosclerosis / drug therapy
  • Atherosclerosis / genetics
  • Atherosclerosis / pathology
  • Atherosclerosis / prevention & control*
  • Atorvastatin / pharmacology*
  • Cholesterol / blood
  • Cholesterol, LDL / blood
  • Drug Therapy, Combination
  • Dyslipidemias / drug therapy
  • Dyslipidemias / genetics
  • Dyslipidemias / pathology
  • Dyslipidemias / prevention & control*
  • Ezetimibe / pharmacology*
  • Gene Expression Regulation / drug effects
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors / pharmacology
  • Intercellular Adhesion Molecule-1 / genetics
  • Occludin / genetics
  • Risk Factors
  • Rosuvastatin Calcium / administration & dosage*
  • Zonula Occludens-1 Protein / genetics

Substances

  • Cholesterol, LDL
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors
  • ICAM1 protein, human
  • OCLN protein, human
  • Occludin
  • TJP1 protein, human
  • Zonula Occludens-1 Protein
  • Intercellular Adhesion Molecule-1
  • Rosuvastatin Calcium
  • Cholesterol
  • Atorvastatin
  • Ezetimibe

Grants and funding

This research was funded by a statutory research granted for the Department of Internal Diseases and Clinical Pharmacology, Medical University of Lodz (number 503/5-165-01/503-51-001-19-00) and by the Medical University of Lodz in cooperation with the European Union (Number 514/6-077-01/514-01-030-02). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.