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.
Highlights
Cardiovascular disease (CVD) is the most common cause of deaths in high-income countries
Zhang et al (2020) [21] report that palmitate decrease ZO-1 and VE-cadherin protein levels in Human umbilical vascular endothelial cells (HUVEC). Another important finding in our study reveals that rosuvastatin (10 μM; 4815 ng/mL; 20 hours) has a positive effect on tight junctions by increasing the expression of the ZO-1 and OCLN genes previously lowered by pre-incubation with 25-OHC (24.83 μM; 10 μg/mL; four hours followed by washout)
Our study indicates that atorvastatin (5 μM; 2793 ng/mL; 20 hours) lowers the level of Intercellular adhesion molecule 1 (ICAM-1) mRNA and protein level in HUVEC prestimulated with 25-OHC (24.83 μM; 10 μg/mL; 4 hours)
Summary
Cardiovascular disease (CVD) is the most common cause of deaths in high-income countries. The onset of atherosclerosis is provoked by a high concentration of oxidized low-density lipoproteins (oxLDLs), which causes a complex inflammatory process, including increased endothelial permeability, monocyte migration, macrophage activation and vascular smooth muscle cell (VSMC) migration into the intima [1]. Gold et al [7] (2012) showed that the inhibition of 25-OHC synthesis attenuates the formation of macrophage foam cells and inhibits atherosclerosis. These findings confirm that 25-OHC is a key element in initiating atherosclerosis and show how important it is to properly understand the effects of 25-OHC on the vascular wall
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
