DJ-1 administration exerts cardioprotection in a mouse model of acute myocardial infarction

FIGURE 1. Effects of systemic DJ-1 administration on infarct size in a mouse model of MI. (A). Representative images of infarcted heart sections. Infarct area is outlined in black. (B). Infarct size quantification after ischemia and ischemia/reperfusion, in the presence and the absence of a systemic administration of DJ-1. Infarct measures are expressed as percentage of left ventricle area. (C). Representative immunohistochemical acquisitions of DJ-1 in the myocardium after ischemia and ischemia/reperfusion, in the presence and the absence of a systemic administration of DJ-1. A sham-operated group was included as baseline. (D). DJ-1 myocardium content quantification across groups. Scale bar: 100 µm; *p < 0.05, Kruskal-Wallis; †p < 0.05 vs. ischemia without DJ-1, ‡p < 0.05 vs. ischemia + DJ-1, #p < 0.05 vs. I/R without DJ-1, Dunn’s test. LV, Left Ventricle; Isch., Ischemia; I/R, Ischemia/Reperfusion.

FIGURE 2. Differential gene expression profile after ischemia/reperfusion, in the presence and the absence of a systemic administration of DJ-1. (A). Heat-map representation of differentially expressed genes. (B). Top ten up- and down-regulated genes after ischemia/reperfusion in the presence of DJ-1 systemic administration. (C). Gprc5a transcript analysis by qPCR. (D). Gene-set enrichment analysis result. Gene-set collections from Wikipathways, Molecular Signature, and Panther databases were considered. *p < 0.05, Mann-Whitney. I/R, Ischemia/Reperfusion; logFC, log-Fold Change.

FIGURE 3. Effects of DJ-1 systemic administration on myocardial leukocyte infiltration after ischemia and ischemia/reperfusion. (A). Representative immunohistochemical acquisitions of myocardial neutrophils (upper panel) and macrophages (lower panel) after ischemia and ischemia/reperfusion in the presence and the absence of a systemic administration of DJ-1. A sham-operated group was included as baseline. (B) and (C) quantification of infiltrated leukocytes. Scale bar: 100 µm; *p < 0.05, Kruskal-Wallis; §p < 0.05 vs. sham, †p < 0.05 vs. ischemia without DJ-1, ‡p < 0.05 vs. ischemia + DJ-1, #p < 0.05 vs. I/R without DJ-1, Dunn’s test. Isch., Ischemia; I/R, Ischemia/Reperfusion.

FIGURE 4. Effects of DJ-1 systemic administration on myocardial apoptosis after ischemia/reperfusion. (A) Casp3 transcript analysis by qPCR. (B) Representative acquisitions of myocardial cleaved Casp-3 immunohistochemistry (upper panel) and TUNEL assay (lower panel) after ischemia/reperfusion. (C) myocardial cleaved Casp-3 immunohistochemistry and TUNEL positive cells quantification. Scale bar: 100 µm; *p < 0.05, Mann-Whitney. I/R, Ischemia/Reperfusion.

FIGURE 5. Systemic DJ-1 administration effects on oxidative stress after ischemia and ischemia/reperfusion. (A) Inos transcript analysis by qPCR. (B) Representative immunohistochemical acquisitions of myocardial iNOS (upper panel) and 8-OHdG (lower panel) after ischemia and ischemia/reperfusion in the presence and the absence of a systemic administration of DJ-1. A sham-operated group was included as baseline. (C) Myocardial iNOS content quantification. (D) Myocardial 8-OHdG content quantification. (E–G) Correlation analyses between iNOS and DJ-1 (E), 8-OHdG and DJ-1 (F), and iNOS and 8-OHdG (G) staining signals. Scale bar: 100 µm; *p < 0.05, Kruskal-Wallis; §p < 0.05 vs. sham, †p < 0.05 vs. ischemia without DJ-1, ‡p < 0.05 vs. ischemia + DJ-1, #p < 0.05 vs. I/R without DJ-1, Dunn’s test; rho, Spearman rank correlation coefficient. Isch., Ischemia; I/R, Ischemia/Reperfusion.