Mouse Mast Cell Protease 4 Deletion Protects Heart Function and Survival After Permanent Myocardial Infarction

Figures

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  FIGURE 1. Kaplan-Meier 28-day survival analysis of mouse survival (Mantel-Cox test) (A) and incidence of left ventricular free wall rupture events (B) following LAD coronary artery ligation (A). PMI: n = 15, SHAM: n = 7.

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  FIGURE 2. Representative cardiac PET imaging of mouse hearts after PMI (A). Mouse hearts were imaged 3 days following surgery with 18F-FDG, and 3-dimension images were reconstructed. Below each reconstructed image is the ECG-gated left ventricular volume, measured through eight time points of the cardiac cycle (gating frames) for the same mouse. Polar maps of the mouse hearts were calculated, with the apex in the center, for the calculation of the infarcted area (B).

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  FIGURE 3. Representative H&E staining of heart sections 3 days following LAD coronary artery ligation (A). § notes eosinophilia sites, ∞ marks inflammatory cell invasion sites. The minimal thickness at the infarct site (B), eosinophilic and inflammatory regions (C) and cardiomyocyte cross-sectional area (D) were measured from the same images and quantified. (#: PMI vs. SHAM, ∗: mMCP-4 KO vs. WT, one-way ANOVA with multiple comparisons Holms-Šidak post-test).

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  FIGURE 4. Representative active caspase-3 (A) and collagen deposition (B) of heart sections following LAD coronary artery ligation or SHAM surgery. Active caspase-3 DAB (brown) staining [quantified in (C)] is visible against the hematoxylin (blue) counterstain, while collagen deposition [quantified in (D)] is marked by the picrosirius red stain. (#: PMI vs. SHAM, ∗: mMCP-4 KO vs. WT, one-way ANOVA with multiple comparisons Holms-Šidak post-test).

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  FIGURE 5. Plasma levels of BNP 1 or 7 days after SHAM or PMI surgery as measured by ELISA. n = 6 (one-way ANOVA P > 0.05).

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  FIGURE 6. Tissue levels of IR-ET-1 1 day after surgery. The levels of immunoreactive ET-1 were measured in the lungs and left heart ventricle. #: PMI vs. SHAM, ∗: WT vs. mMCP-4 KO, n = 5–7, one-way ANOVA with multiple comparisons Holms-Šidak post-test).

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  FIGURE 7. Analysis of the in vitro hydrolysis of recombinant mouse IGF-1 (LC/MS/MS) and Big ET-1 (HPLC) by recombinant mMCP-4, with the chymase inhibitor TY-51469 (10 μM). In (A), the results of recombinant mMCP-4 activity on mouse IGF-1 are shown using trypsin-degradation products, with GFYFNKPTGYGSSIR coming from the complete IGF-1 sequence and GFYFNKPTGY coming from the mMCP-4-cleaved sequence. In (B), the area under the curve (AUC) was measured for Big ET-1 and ET-1 (1–31) using HPLC at 214 nm detection. n = 3 (one-way ANOVA with multiple comparisons Holms-Šidak post-test). In (C), the complete sequence of mouse IGF-1 is shown, with the disulfide bonds shown with brackets, the analyzed trypsin-digested product in bold and the cleavage site in increased type size.