P6297Genetic mutation of galectin-3 altered the temporal evolution of macrophage polarization and healing affecting the post myocardial infarct remodeling in mice
Authors: Gonzalez G , Cassaglia P D , Penas F , Martinez N L , Bettazza C , Wilensky L , Fontana Estevez F , Noli Truant S , Miksztowicz V , Cevey A , Cicale E , Berg G , Fernandez M , Goren N , Morales C 
Published: October 2019

Background

Myocardial infarction (MI) is a dynamic process that leads to ventricular remodeling (VR) and largely to heart failure (HF). Previous studies established that Galectin-3 (Gal-3) is highly increased in the infarct zone from the beginning of MI and also that it is a prognostic marker of HF.

Purpose

We aimed to study the effects of genetic deletion of Gal-3 on macrophage (MΦ) infiltration, cytokines expression, fibrosis and MMP-2 activity as well as VR and function after MI in mice.

Methods

Male C57BL/6J and Gal-3 KO mice were subjected to permanent coronary ligature or sham. At 1 week post-MI LV function and VR were studied by echocardiography. We also studied in the infarct zone: 1) F4/80+ MΦ infiltration by flow cytometry; 2) M2 macrophage polarization by detection of mannose receptor (MR) and chitinase-3-like protein-3 (YM1) phenotype markers by rt-qPCR; 3) mRNA expression of TNF-α, IL-6, IL-10 and TGF-β; 4) MMP-2 activity by zymography and 5) fibrosis by histology.

Results

Results are expressed as X±SEM; *p<0.05 C57 MI vs Gal-3 KO MI. After 1 week post-MI, the pulmonary congestion assessed by the lung weight/body weight ratio (mg/g) was 9±0.4, 8±0.5 and 9±0.4 in C57 sham, Gal-3 KO sham and C57 MI, respectively, nevertheless it was severely increased to 15±1.2* in infarcted Gal-3 KO mice. MΦ infiltration, cytokine expression and MMP-2 activity in the infarct zone are shown in the table.

Table 1 Groups F4/80+ MR YM1 TNF-α IL-6 IL-10 TGF-β MMP-2 MΦ (%) (A.U.) (A.U.) (A.U.) (A.U.) (A.U.) (A.U.) (A.U.) C57 MI 5.6±0.9 (8) 0.5±0.2 (3) 1.7±0.4 (3) 26±0.3 (3) 1.7±0.5 (3) 0.7±0.03 (3) 1.8±0.5 (3) 1±0.1 (5) Gal-3 KO MI 2.6±0.4* (8) 1.8±0.4*(8) 4.2±0.4* (8) 0.4±0.5* (8) 6.7±0.9 * (8) 2.8±0.3 * (8) 0.5±0.2 * (8) 2±0.4* (6) A.U.: Arbitrary units. In MR, YM1, TNF-α, IL-6, IL-10 and TGF-β represent mRNA expression. In MMP-2 represent gelatinolytic activity. Number of samples is shown between parentheses.

After 1 week post-MI, LV end diastolic dimension was increased from 4.4±0.1 to 4.8±0.2* at the same time that ejection fraction (%) was significantly reduced from 47±2 to 38±3* in C57+MI (13) and Gal-3 KO+MI (16) respectively. Collagen concentration in the infarct zone was significantly reduced from 30±1.2% (6) to 17±0.5* % (8) in C57 and Gal-3 KO respectively.

Conclusion(s)

Gal-3 is an essential regulatory factor for the early wound healing since it regulates the dynamics of the reparative process through the phenotypic profile of MΦ, the pro- and anti-inflammatory cytokines expression and fibrosis along the temporal evolution of MI in mice. The deficit of Gal-3 diminished the infiltration of MΦ altering its phenotypic polarization and consequently, the dynamics of the wound healing as well as aggravating the functional and structural evolution of cardiac remodeling.

Acknowledgement/Funding

Argentine Agency for Promotion of Science and Technology (PICT 2014-2320), University of Buenos Aires (UBACyT 20020170100619BA)