Fig 1.
Rosuvastatin inhibits the formation of atherosclerotic plaques in ApoE-/- mice.
(A) Representative en face images of Sudan IV-stained aortas from ApoE-/- mice treated with PBS or rosuvastatin (5 mg/kg/2 days) followed by a HFD for 12 weeks. The mean aortic lesion area was quantified. Bar = 2 mm. (n ≥ 6 mice per group). ****P < 0.0001 by unpaired Student’s t test. (B-C) Representative Oil Red O-stained aortic root sections with quantification of lesion size. Scale bar = 100 μm. **P < 0.01 by unpaired Student’s t test. (D) Body weight of each group. *P < 0.05, ***P < 0.001 by unpaired Student’s t test. (E-F) Serum TC and LDL-C levels in each group of mice. *P < 0.05 by unpaired Student’s t test.
Fig 2.
Rosuvastatin prevents the impairment of endothelial cell activity induced by ox-LDL.
(A) Effects of different concentrations of rosuvastatin (0.1, 1, 5 and 10 µM) on HUVEC viability for 24 h. (B) Effects of treatment with different concentrations of ox-LDL (50, 100 and 200 µg/mL) for 24 h on HUVEC viability. (C) HUVECs were treated with different concentrations of rosuvastatin and ox-LDL (200 µg/ml) for 24 h. *P < 0.05, ***P < 0.001 by one-way ANOVA. (D) Bar chart showing the signaling pathways enriched with DEGs in the RNA-Seq dataset (GSE206927) of ox-LDL-treated HUVECs according to GO analysis. (E) Bubble chart showing the signaling pathways enriched with DEGs according to KEGG analysis. (F-G) HUVECs stimulated with ox-LDL and different concentrations of rosuvastatin were stained with DAPI (blue) and Ki67 (purple). Ki67-positive cells were quantified, bar = 100 μm, *P < 0.05, ****P < 0.0001 by one-way ANOVA.
Fig 3.
Rosuvastatin inhibits ox-LDL-induced oxidative stress in endothelial cells.
HUVECs were treated with ox-LDL in the presence or absence of different concentrations of rosuvastatin (0.1, 1, 5 and 10 µM) for 24 h. (A) NO production in HUVECs. (B) eNOS mRNA expression in HUVECs. (C) A microplate reader was used to measure the fluorescence intensity of the ROS at an excitation wavelength of 488 nm and an absorption wavelength of 525 nm via a fluorescent probe DCFH-DA kit, and Rosup was used as a positive control. (D) The mean intracellular fluorescence intensity was analyzed via fluorescence microscopy. The data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, **P < 0.0001 by one-way ANOVA.
Fig 4.
Rosuvastatin inhibits ox-LDL-induced apoptosis of endothelial cells.
(A-B) Bcl2 and Bax mRNA expression in HUVECs treated with different concentrations of rosuvastatin and ox-LDL (200 µg/ml) for 24 h. **P < 0.01, ***P < 0.001, ****P < 0.0001 by one-way ANOVA. (C-D) BCL-2 and Bax protein expression in HUVECs treated with different concentrations of rosuvastatin and ox-LDL (200 µg/ml) for 24 h. The data are presented as the means ± SEMs. *P < 0.05 by one-way ANOVA. (E) HUVECs stimulated with ox-LDL and different concentrations of rosuvastatin were stained with DAPI (blue), Bax (green) and mitochondria (red); scale bar = 20 μm. (F) Early and late apoptosis of HUVECs treated with 100 µg/mL ox-LDL and 10 μmol/L rosuvastatin for 24 h. The quantification results are shown on the right (n = 5). ***P < 0.001, ****P < 0.0001 by one-way ANOVA.
Fig 5.
Rosuvastatin inhibits the NF-κB signaling pathway in endothelial cells.
(A-D) Protein levels of IkBα, p-IkBα, P65 and p-P65 in HUVECs treated with or without 10 µM rosuvastatin and treated with 100 µg/mL ox-LDL for 24 h. The data are presented as the means ± SEMs. *P < 0.05, **P < 0.01 by one-way ANOVA. (E) Schematic diagram illustrating the role of rosuvastatin in ox-LDL-induced endothelial cell dysfunction. Rosuvastatin regulates oxidative stress and apoptosis-related gene transcription in endothelial cells by inhibiting ox-LDL-induced IKBα and P65 activation in endothelial cells.