Interleukin-12p35 deficiency enhances mitochondrial dysfunction and aggravates cardiac remodeling in aging mice

Figures

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  Figure 1. Effects of IL-12p35 KO on cardiac function. (A). Representative M-mode echocardiograms, (B). LVPWT, (C). IVSD, (D). LVESD, (E). LVEF and (F). FS for each group. * p<0.05 vs. the young WT group; # p<0.05 vs. the aged WT group; n=11-12 for each group.

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  Figure 2. Regulatory role of IL-12p35 KO in cardiac remodeling. (A–C). Body weights (BW), heart weights (HW) and HW/BW ratios were measured in the four groups; n=10 for each group. (D). The cross-sectional areas (CSA) of cardiomyocytes and cardiac fibrotic areas were measured by WGA staining and Masson staining, respectively (200x). * p<0.05 vs. the young WT group; # p<0.05 vs. the aged WT group; n=5 for each group.

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  Figure 3. Effects of IL-12p35 KO on the expression of aging marker-related proteins. (A) The left ventricular p16, p21, p53 and Sirt1 expression levels in each group were investigated by Western blotting analysis; n=10-11 for each group. (B) Cardiac p53 expression was detected by immunofluorescence staining (200x); n=5 for each group. * p<0.05 vs. the young WT group; # p<0.05 vs. the aged WT group.

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  Figure 4. Mitochondrial fluorescence intensity of calcium and ROS and membrane potentials were detected. (A). The mitochondrial ROS fluorescence intensity was investigated by the Flow Cytometry analyses. (B). The MitoSOX Red Mitochondrial Superoxide Indicator probe and the Rhod-2 AM probe were used to mark mitochondrial calcium ions and ROS, respectively (400x). (C). The mitochondrial membrane potential was detected using the JC-1 probe (400x). N=5-6 for each group.

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  Figure 5. Effects of IL-12p35 KO on mitochondrial dysfunction. (A, B). Cardiac 8-OHdg expression (200x) and mitochondrial 8-OHdg levels were detected. (C). The activity of mitochondrial enzyme complexes I, II, III and IV in the four groups was measured. (D). The mitochondrial ATP levels in each group were detected. * p<0.05 vs. the young WT group; # p<0.05 vs. the aged WT group; n=4-5 for each group.

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  Figure 6. Effects of IL-12p35 KO on cardiomyocyte apoptosis. (A) Mitochondrial AIF expression and nuclear AIF and Cle-PARP expression were measured by Western blotting analysis. (B) Cardiac AIF expression was detected by immunofluorescence staining (200x). (C) TUNEL staining was performed to mark the apoptotic cells and the positive cell numbers in each group (200x). * p<0.05 vs. the young WT group; # p<0.05 vs. the aged WT group; n=5 for each group.