Table 1.
Morphological parameters obtained by coronary angiography and optical coherence tomography.
Fig 1.
Images of optical coherence tomography (OCT).
OCT images are shown for each group. Scale bars = 1 mm. ARB, angiotensin II receptor blocker.
Fig 2.
Morphologic changes by histological analysis.
(A) Tissues were stained with hematoxylin & eosin (H&E) and Masson’s trichrome. Alpha-smooth muscle actin (α-SMA) content of the plaques is shown by immunohistochemical staining of α-SMA, and collagen content of the plaques is represented by Sirius red staining visualized under polarized light. (B) Intima/plaque percentage for vessels in each rabbit group. (C) Lipid deposition content of plaques analyzed by Oil Red O staining. Higher magnification of regions within the black boxes in (A). (D) Relative intensity of Oil Red O staining (lipid deposition percentage) for each group. Scale bars = 100 μm. Data are mean±SEM. * p<0.05 vs. positive control group and † p<0.05 vs. statin group. ARB, angiotensin II receptor blocker.
Fig 3.
Immunohistochemical staining of macrophage markers.
(A) Tissues immunologically stained with RAM11 and percentage area. (B) Tissues immunologically stained with TNF-α and percentage area. (C) Tissues immunologically stained with CD163 and percentage area. (D) Tissues immunologically stained with CD206 and percentage area. Scale bars = 100 μm. Data are mean±SEM. * p<0.05 vs. positive control group. ARB, angiotensin II receptor blocker; CD163, cluster of differentiation 163; CD206, cluster of differentiation 206; TNF-α, tumor necrosis factor-α.
Fig 4.
Macrophage polarization assessed by confocal immunofluorescence microscopy.
(A) Confocal immunofluorescence microscopy showing CD68 (green), iNOS (red), and Arg-1 (white) localization in the same vessel. Blue represents 4′,6-diamidino-2-phenylindole staining of nuclei. (B) Relative area of CD68–iNOS staining in each group. (C) Relative area of CD68–Arg-1 staining in each group. Data are mean±SEM. * p<0.05 vs. positive control group, † p<0.05 vs. statin group, and ‡ p<0.05 vs. ARB group. ARB, angiotensin II receptor blocker; Arg-1, arginase-1; iNOS, inducible nitric oxide synthase.
Fig 5.
Reverse transcription–polymerase chain reaction (RT-PCR) and Western blot analysis in rabbit vessels.
(A) RT-PCR expression of TNF-α, RAGE, HMG CoA reductase, iNOS, AGTR1, and Arg-1. (B-G) Comparisons of relative mRNA expression, normalized to expression of GAPDH as the housekeeping gene. (H) Western blot expression of TNF-α, RAGE, iNOS, and Arg-1. (I-L) Representative data showing protein expression, normalized to expression of GAPDH as the housekeeping gene. Data in the bar graphs are quantified ratios of the signal for TNF-α, RAGE, HMG CoA reductase, AGTR1, iNOS, and Arg-1 relative to the signal for GAPDH, presented as fold increases. Data are mean±SEM. * p<0.05 vs. positive control group, † p<0.05 vs. statin group, and ‡ p<0.05 vs. ARB group. AGTR1, angiotensin II receptor type 1; ARB, angiotensin II receptor blocker; Arg-1, arginase-1; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HMG CoA, 5-hydroxy-3-methylglutaryl–coenzyme A; iNOS, inducible nitric oxide synthase; RAGE, receptor for advanced glycation end products; TNF-α, tumor necrosis factor-α.
Fig 6.
Effects of statin and ARB on the macrophage RAW 264.7 cell line, with cytokine mRNA and protein expressions.
RAW 264.7 cells were stimulated with 0.1 μg LPS for 4 h, followed by 2 mM ATP for 2 h. PBS was used as the positive control. After stimulation, cells were treated for 6 h (A, C) or 24 h (B, D) with 10 μM statin, 10 μM ARB, or the combination (10 μM statin + 10 μM ARB). Data in bar graphs are quantified ratios presented as fold increases. Data represent n = 6. * p<0.05 vs. LPS group and † p<0.05 vs. LPS+statin group. ARB, angiotensin II receptor blocker; Arg-1, arginase-1; combi, combination of statin and ARB; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; IL-1β, interleukin 1 beta; IL-6, interleukin 6; iNOS, inducible nitric oxide synthase; LPS, lipopolysaccharide; mIL-1β, mature IL-1β; PBS, phosphate-buffered saline; pIL-1β, pro IL-1β; TNF-α, tumor necrosis factor-α.
Fig 7.
Schematic diagram showing synergistic protective effects of statin and ARB on favorable macrophage polarization.