Fig 1.
FGF-9 Inhibits Infarct and Fibrosis in the Post-MI Diabetic Myocardium.
A-F: Representative images of Masson’s trichrome stained sections are shown depicting infarct (A-C) and interstitial fibrosis (D-F) for control and experimental groups 2 weeks post-MI. Single arrows represent infarcted myocardium. Double arrows represent interstitial fibrosis. G: Histogram of quantitative infarct size analysis suggests a significant decrease in infarct size in the MI+FGF-9 group compared to the MI group. H: Histogram of mean left ventricular cardiac myocyte cross sectional interstitial fibrotic (IF) area (mm2). n = 5–8 animals/group. *p<0.05 vs. sham and #p<0.05 vs. MI.
Fig 2.
FGF-9 Blunts Monocyte Infiltration in Infarcted Diabetic Myocardium.
A-O: Representative photomicrographs illustrate CD14 positive monocytes in red (A, F, and K), cardiac myocytes in green (B, G, and L), total nuclei in blue (C, H, and M), merged images (D, I, and N), and enhanced merged images (E, J, and O) for all control and experimental groups. White arrows are used to show the areas enhanced in D, I, and N. Scale bar = 75μm. P: Histogram of quantitative monocyte infiltration data 2 weeks post-MI. n = 5 animals/group. *p<0.05 vs. sham and #p<0.05 vs. MI.
Fig 3.
Monocyte to M1 Macrophage Polarization is Blunted Following FGF-9 Treatment.
A-O: Representative photomicrographs demonstrate iNOS positive M1 macrophages in red (A, F, and K), cardiac myocytes in green (B, G, and L), total nuclei in blue (C, H, and M), merged images (D, I, and N), and enhanced merged images (E, J, and O) for all control and experimental groups. White arrows are used to show the areas enhanced in D, I, and N. Scale bar = 75μm. P: 2 weeks post-MI, M1 macrophage popoluations are significantly decreased in the MI+FGF-9 group. n = 5 animals/group. Q: Pro-inflammatory TNF-α mRNA expression was evaluated by RT-PCR and data provided suggest a significant increase post-MI. R: Proinflammatory IL-6 mRNA expression was significantly decreased following FGF-9 treatement in the post-MI diabetic heart. *p<0.05 vs. sham and #p<0.05 vs. MI.
Fig 4.
M2 Macrophages are Significantly Enhanced Following FGF-9 Treatment.
A-O: Representative photomicrographs demonstrate CD206 positive M2 macrophages in red (A, F, and K), cardiac myocytes in green (B, G, and L), total nuclei in blue (C, H, and M), merged images (D, I, and N), and enhanced merged images (E, J, and O) for all control and experimental groups. White arrows are used to show the areas enhanced in D, I, and N. Scale bar = 75μm. P: Quantitative data suggest that M2 macrophage concentrations are significantly greater in the MI+FGF-9 group relative to sham and MI groups. n = 5 animals/group. Q: Anti-inflammatory IL-10 mRNA expression, evaluated by RT-PCR, was significantly upregulated post-FGF-9 treatment. *p<0.05 vs. sham and #p<0.05 vs. MI.
Fig 5.
FGF-9 Treatment Augments Pro- and Anti-Inflammatory Cytokine Secretion.
2 weeks following MI, heart homogenates were used to quantify pro- and anti-inflammatory cytokine secretion via ELISA analysis. Pro-inflammatory cytokine expression, including that of TNF-α, MCP-1, and IL-6, are shown in A, B, and C, respectively whereas anti-inflammatory IL-10 and IL-1RA cytokine expression are depicted in D and E, respectively. n = 5 animals/group. *p<0.05 vs. sham and #p<0.05 vs. MI.
Fig 6.
Exogenous FGF-9 Treatment Improves Cardiac Function Following MI.
2 weeks post MI, 2D transthoracic echocardiography was performed on control and experimental animals. All mean data for each quantified measurement are presented in A. Quantitative analyses are shown for left ventricular internal dimension-diastole (LVIDd) (B), left ventricular internal dimension-systole (LVIDs) (C), fractional shortening (FS) (D), left ventricular volume at end diastole (EDV) (E), left ventricular volume at end systole (ESV) (F), and ejection fraction (EF) (G). n = 5 animals/group. *p<0.05 vs. sham and #p<0.05 vs. MI.