miR-199a-5p Relieves Obstructive Sleep Apnea Syndrome-Related Hypertension by Targeting HIF-1α

Figures

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  Figure 1. miR-199a-5p and HIF-1α expression in the animal model of OSAS with hypertension. mRNA expression of miR-199a-5p in (a) myocardial tissue and (b) aortic tissue by qRT-PCR. mRNA expression of HIF-1α in (c) myocardial tissue and (d) aortic tissue by qRT-PCR. Protein expression of HIF-1α and quantification in (e, f) myocardial tissue and (g, h) aortic tissue by western blot. CIH: chronic intermittent hypoxia. ∗∗P < 0.01, ∗∗∗P < 0.001.

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  Figure 2. Overexpressed miR-199a-5p reduces blood pressure and relieves oxidase stress and inflammation. (a) Blood pressure in rats with exposure to CIH during 56 days. (b) SOD, (c) MDA, (d) hs-CRP, and (e) sE-s in serum of rats in control, CIH, CIH+NC, and CIH+miR-199a-5p groups. CIH: chronic intermittent hypoxia. ∗P < 0.05, ∗∗P < 0.01.

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  Figure 3. Overexpressed miR-199a-5p attenuated vascular remodeling. (a) Representative H&E staining and (b) Masson staining in the heart and abdominal aorta of rats in control, CIH, CIH+NC, and CIH+miR-199a-5p groups. Scale bar = 25 μm. The fibrosis area of Masson staining in the (c) heart and (d) abdominal aorta. The collagen deposition was stained in blue. CIH: chronic intermittent hypoxia; NC: negative control. ∗∗P < 0.01, ∗∗∗P < 0.001. n = 6 per group.

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  Figure 4. Overexpressed miR-199a-5p attenuated vascular remodeling. (a) mRNA expression of miR-199a-5p in myocardial tissue by qRT-PCR. (b) Protein expression of HIF-1α, TGFβ1, and α-SMA in myocardial tissue by western blot and (c) quantification. (d) mRNA expression of miR-199a-5p in aortic tissue by qRT-PCR. (e) Protein expression of HIF-1α, TGFβ1, and α-SMA in aortic tissue by western blot and (f) quantification. CIH: chronic intermittent hypoxia; NC: negative control. ∗P < 0.05, ∗∗P < 0.01 compared with control; #P < 0.05, ##P < 0.01, and ###P < 0.001 compared with CIH+NC.

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  Figure 5. miR-199a-5p targeted HIF-1α. mRNA expression of (a) miR-199a-5p and (b) HIF-1α in A7r5 cells under normoxia and hypoxia (1% O2) by qRT-PCR. (c) Protein expression of HIF-1α in A7r5 cells by western blot and (d) quantification. (e) Luciferase reporter assay to validate the putative (f) miR-199a-5p-binding site in the 3′-UTR of HIF-1α. WT: wild type; Mut: mutant; NC: negative control. ∗P < 0.05, ∗∗P < 0.01.

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  Figure 6. miR-199a-5p/HIF-1α inhibited proliferation of vascular smooth muscle cells under hypoxia. mRNA expression of (a) miR-199a-5p and (b) HIF-1α in A7r5 cells under normoxia and hypoxia (1% O2) by qRT-PCR. (c) Protein expression of HIF-1α in A7r5 cells by western blot and (d) quantification. (e) BrdU (red) incorporation in A7r5 cells transfected with the miR-199a-5p mimic, miR-199a-5p inhibitor, and corresponding controls under hypoxia (1% O2). Scale bar = 50 μm. (f) Quantification of BrdU-positive cell number. (g) Colony formation assay in A7r5 cells transfected with the miR-199a-5p mimic, miR-199a-5p inhibitor, and corresponding controls under hypoxia (1% O2). Scale bar = 50 μm. (h) Quantification of relative number of colonies. NC: negative control. ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.