Effects of progranulin on the pathological conditions in experimental myocardial infarction model

Figures

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  Figure 1. Progranulin expression in the heart after myocardial infarction in mice. (A) Representative images show progranulin (58–68 kD) in the ischemic area 6 h and 1, 3, 5, and 7 days after permanent occlusion of left coronary artery (LCA) by Western blotting. (B) Quantitative analysis of progranulin normalized to GAPDH in the ischemic area. (C) Representative images show progranulin (58–68 kD) in non-ischemic area 6 h and 1, 3, 5, and 7 days after permanent occlusion of LCA by Western blotting. (D) Quantitative analysis of progranulin normalized to GAPDH in non-ischemic area. The expression of progranulin increased in the ischemic area 1, 3, and 5 days after permanent occlusion of LCA. Data are the means ± SEM. (n = 5–7) *p < 0.05, **p < 0.01 versus sham-operated group (one-way ANOVA followed by Dunnett's test).

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  Figure 2. Localization of upregulated progranulin and exploration of progranulin expressed cells after myocardial infarction in mice. (A) Representative images show immunostaining for progranulin in sham-operated group, and at infarct, border and remote areas 1 day after permanent occlusion of LCA. Nuclei were stained with Hoechst 33342. Scale bar: 500 μm. (B) Scheme of heart section indicating the infarct, border, and remote areas after permanent occlusion of LCA. (C) Quantitative analysis of fluorescence intensity of progranulin. Progranulin expression significantly increased in the border area 1 day after permanent occlusion of LCA. (D) Representative images at ×60 and ×600 magnification show immunostaining for progranulin and NIMP-R14 at the border area 1 day after permanent occlusion of LCA. Scale bar at the ×60 and ×600 magnification: 50 μm and 5 μm representatively. Data are the means ± SEM. (n = 5–6) *p < 0.05 versus sham-operated group (one-way ANOVA followed by Dunnett's test).

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  Figure 3. Administration of recombinant progranulin decreased infarct size after myocardial infarction in mice. (A) Representative photographs show the heart sections stained by 2, 3, 5-Triphenyl tetrazolium chloride (TTC) solution, 24 h after permanent occlusion of LCA. (B) Quantitative analysis of infarct size 24 h after permanent occlusion of LCA. The administration of recombinant progranulin significantly decreased myocardial infarct size compared with vehicle-treated group. Data are the means ± SEM. (n = 7–9) *p < 0.05 vs. vehicle-treated group (two-tailed Student's t-test).

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  Figure 4. Administration of recombinant progranulin suppressed the infiltrating neutrophils after myocardial infarction in mice. (A) Reparative images show immunostaining for NIMP-R14 at the infarct and border areas 1 day after permanent occlusion of LCA. Nuclei were stained with Hoechst 33342. (B) Number of neutrophils per mm2 at infarct area after permanent occlusion of LCA. (C) Number of neutrophils per mm2 at border area after permanent occlusion of LCA. The administration of recombinant progranulin reduced the number of infiltrating neutrophils at the infarct area compared with vehicle-treated group. Scale bar: 50 μm. Data are the means ± SEM. (n = 5–6) *p < 0.05, **p < 0.01 versus sham-operated group (two-tailed Student's t-test), †p < 0.05 vs. vehicle-treated group (one-tailed Student's t-test).

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  Figure 5. Administration of recombinant progranulin decreased deterioration of cardiac dysfunction after myocardial ischemia–reperfusion injury in rabbits. (A) The positive values of the first derivative in left ventricular pressure (dP/dt) and (B) the negative dP/dt were determined by cardiac catheterization 2 weeks after myocardial ischemia/reperfusion (I/R). (C) Left ventricular internal diameter at diastole (LVIDd), (D) left ventricular internal diameter at systole (LVIDs), (E) ejection fraction (EF), and (F) fractional shortening (FS) were measured by echocardiography analysis. The administration of recombinant progranulin significantly ameliorated the deterioration of + dP/dt, LVIDd, LVIDs, EF and FS compared with vehicle-treated group. Data are the means ± SEM. (n = 3–7) *p < 0.05 versus vehicle-treated group (A, B; two-tailed Student's t-test, C–F; one-tailed Student's t-test).

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  Figure 6. Administration of recombinant progranulin decreased myocardial fibrosis size after myocardial ischemia–reperfusion in rabbits. (A) Representative photographs show LV cross-sections stained by Masson’s trichrome solutions 2 weeks after myocardial I/R. (B) Quantitative analysis of fibrosis size 2 weeks after myocardial I/R. The administration of recombinant progranulin significantly reduced fibrosis size compared with vehicle-treated group. Scale bar: 2.5 mm. Data are the means ± SEM. (n = 3–7) *p < 0.05 versus vehicle-treated group (one-tailed Student's t-test).