Therapeutic silencing of microRNA-33 inhibits the progression of atherosclerosis in Ldlr-/- mice--brief report - PubMed (original) (raw)

Therapeutic silencing of microRNA-33 inhibits the progression of atherosclerosis in Ldlr-/- mice--brief report

Noemi Rotllan et al. Arterioscler Thromb Vasc Biol. 2013 Aug.

Abstract

Objective: To study the efficacy of anti-miRNA-33 therapy on the progression of atherosclerosis.

Approach and results: Ldlr(-/-) mice were injected subcutaneously with PBS, control, or anti-miR-33 oligonucleotides weekly and fed a Western diet for 12 weeks. At the end of treatment, the expression of miR-33 target genes was increased in the liver and aorta, demonstrating effective inhibition of miR-33 function. Interestingly, plasma high-density lipoprotein (HDL)-cholesterol was significantly increased in anti-miR-33-treated mice but only when they were fed a chow diet. However, HDL isolated from anti-miR-33-treated mice showed an increase cholesterol efflux capacity compared with HDL isolated from nontargeting oligonucleotide-treated mice. Analysis of atherosclerosis revealed a significant reduction of plaque size and macrophage content in mice receiving anti-miR-33. In contrast, no differences in collagen content and necrotic areas were observed among the 3 groups.

Conclusions: Long-term anti-miR-33 therapy significantly reduces the progression of atherosclerosis and improves HDL functionality. The antiatherogenic effect is independent of plasma HDL-cholesterol levels.

Keywords: ABCA1 protein; atherosclerosis; cholesterol, HDL; macrophages; miR-33.

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Figures

Figure 1

Figure 1. Lipid analysis and gene expression in _Ldlr_−/− mice injected with PBS, Ctrl ASO or miR-33 ASO during a progression study

A) Experimental outline of PBS, Ctrl ASO or miR-33 ASO treatment of _Ldlr_−/− mice fed a WD for 12 weeks. B–C) Hepatic mRNA (left panel) and protein (right panel) expression of miR-33 target genes from mice treated with PBS, Ctrl ASO and miR-33 ASO for 12 weeks and fed a WD. *Indicates p< 0.05 compared with miR-33 ASO with PBS and control ASO group. D–F) Total cholesterol (D), HDL-C (E) and triglyceride (F) levels at starting point (0 week) and at 4 and 12 weeks on WD and PBS, Ctrl ASO and miR-33 ASO treatment. All of the data represent the mean ± SEM; (PBS n=5, control ASO n=9 and miR-33 ASO n=9). G) Cholesterol lipoprotein profile from pooled plasma from _Ldlr_−/− mice injected with PBS, Ctrl ASO and miR-33 ASO after 12 weeks on WD.

Figure 2

Figure 2. Reduction in lesion area and gene expression of inflammatory, adhesion, matrix and macrophage markers from RNA of total aortas in _Ldlr_−/− mice injected with PBS, Ctrl ASO or miR-33 ASO during a progression study

A–D) Representative histological analysis of cross-sections from the aortic sinus stained with Oil-red-O (ORO) (A), hematoxylin and eosin (H&E) (B), Masson’s trichrome (MT) (C), and CD68, α-SMC actin (D). Quantification of the lesion area, fibrous cap, necrotic core and macrophage content are represented in the right panels. All of the data represents the mean ± SEM; (PBS n=5, control ASO n=9 and miR-33 ASO n=9). *Indicates p<0.05 compared to miR-33 ASO with PBS and Ctrl ASO group. E) Expression profile of atherosclerotic-related genes assessed by real-time qPCR. Five independent qPCR reactions were carried out for each condition. The fold change for each gene of mice treated with miR-33 ASO compared to mice treated with Ctrl ASO. The data represents the mean ± SEM. *Indicates p< 0.05 compared to the miR-33 ASO with the Ctrl ASO group. Bar scale = 400μm. F) Representative Western blot analysis of ABCA1 and HSP90 expression from aortic lysates of mice treated with Ctrl ASO and miR-33 ASO.

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