miR‐1929‐3p Overexpression Alleviates Murine Cytomegalovirus‐Induced Hypertensive Myocardial Remodeling by Suppressing Ednra/NLRP3 Inflammasome Activation

Figures

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  Figure 1. MCMV infection raised BP and decreased the expression of miR-1929-3p in C57BL/6 mice. (a) Schematic of intervention here and in Figure 2 in the experimental group. (b) PCR analysis of the MCMV IE gene in the myocardium of C57BL/6 mice. (c) ELISA detection of IgG concentration in plasma. (d) ELISA detection of IgM concentration in plasma. (e) qRT-PCR analysis of miR-1929-3p expression in the myocardium. The value was normalized to the value observed with the control groups, which was set to 1: (f) SBP; (g) DBP; (h) MAP. The data are expressed as the means ± SEM (n = 5), ∗P < 0.05, the MCMV groups vs. the age-matched control groups. #P < 0.05, the MCMV 9-month groups vs. the control 7-month groups.

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  Figure 2. MCMV infection induced myocardial remodeling in mice. (a) Determination of HW/BW%. (b) Determination of LVW/BW%. (c) Representative images of H&E staining in the heart. (d) Quantitative analysis of the cell size (μm2) of cardiomyocytes in all groups. (e) Representative images of Masson's trichrome stain in the heart. (f) Quantitative analysis of fibrosis in all groups. The data are expressed as the means ± SEM (n = 5), ∗P < 0.05, the MCMV groups vs. the age-matched control groups. HW/BW%: heart-to-body weight ratio; LVW/BW%: left ventricle-to-body weight ratio. Images were magnified to ×400 power, scale bars = 20 μm.

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  Figure 3. miR-1929-3p overexpression relieved MCMV-induced myocardial remodeling in mice. (a) Schematic of intervention here and in Figures 4–7 in the experimental group. (b) The expression of miR-1929-3p in the myocardium. The value was normalized to the respective control groups, which was set to 1: (c) SBP; (d) DBP; (e) MAP; (f) HW/BW%; (g) LVW/BW%. (h) Representative images of H&E staining. (i) Quantitative analysis of the cell size (μm2) of cardiomyocytes in all groups. (j, k) qRT-PCR analysis of cardiac-specific fetal genes BNP and β-MHC. The value was normalized to the respective control groups, which was set to 1. (l) Representative images of Masson's trichrome stain. (m) Quantitative analysis of fibrosis in all groups. The data are expressed as the means ± SEM (n = 5), ∗P < 0.05 vs. the age-matched control groups. #P < 0.05, the MCMV+miR-1929-3p groups vs. the age-matched MCMV groups. Images were magnified to ×400 power; scale bars = 20 μm.

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  Figure 4. miR-1929-3p overexpression downregulated Ednra and alleviated MCMV-induced cardiac dysfunction. (a) Western blotting analysis of Ednra protein. (b) Densitometric analysis of (a). (c) qRT-PCR analysis of Ednra mRNA. The value was normalized to the control groups, which was set to 1. (d) Images of the M-mode of LV. (e–i) LVPWd, LVIDs, LVIDd, EF%, and FS%. The data are expressed as the means ± SEM (n = 5), ∗P < 0.05 vs. the age-matched control groups. #P < 0.05, the MCMV+miR-1929-3p groups vs. the MCMV groups. LVPWd: left ventricular end-diastolic posterior wall dimension; LVIDs: left ventricular end-systolic diameter; LVIDd: left ventricular end-diastolic diameter; EF%: ejection fraction; FS%: fractional shortening.

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  Figure 5. miR-1929-3p overexpression reduced MCMV-induced expression of the NLRP3 inflammasome of the myocardium in morphology. (a, c, e) Immunohistochemical analysis of NLRP3, ASC, and caspase-1. (b, d, f) Densitometric analysis of (a), (c), and (e). The data are expressed as the means ± SEM (n = 5), ∗P < 0.05 vs. the age-matched control groups. #P < 0.05, the MCMV+miR-1929-3p groups vs. the age-matched MCMV groups. Images were magnified to ×400 power; scale bars = 20 μm.

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  Figure 6. miR-1929-3p overexpression suppressed MCMV-induced expression of the NLRP3 inflammasome and secretion of IL-1β in the myocardium after MCMV infection. (a) Western blotting analysis of NLRP3, ASC, pro-caspase-1, caspase-1, pro-IL-1β, IL-1β, and IL-18 protein. (b–j) Densitometric analysis of (a). The data are expressed as the means ± SEM (n = 5), ∗P < 0.05 vs. the age-matched control groups. #P < 0.05, the MCMV+miR-1929-3p groups vs. the age-matched MCMV groups.

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  Figure 7. miR-1929-3p overexpression decreased ET-1 and IL-1β activation in MCMV-infected mice. (a, c, e) ELISA detection of ET-1, IL-1β, and IL-18 concentration in plasma. (b, d, f) ELISA detection of ET-1, IL-1β, and IL-18 concentration in cardiac tissues. The data are expressed as the means ± SEM (n = 5), ∗P < 0.05 vs. the age-matched control groups. #P < 0.05, the MCMV+miR-1929-3p groups vs. the age-matched MCMV groups. (g) A schematic model showing the essential role of miR-1929-3p and Ednra in MCMV-induced hypertensive cardiac remodeling.