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. 2015 Jul:84:24-35.
doi: 10.1016/j.yjmcc.2015.04.003. Epub 2015 Apr 11.

Resolvin D1 activates the inflammation resolving response at splenic and ventricular site following myocardial infarction leading to improved ventricular function

Vasundhara Kain  1 , Kevin A Ingle  1 , Romain A Colas  2 , Jesmond Dalli  2 , Sumanth D Prabhu  1 , Charles N Serhan  2 , Medha Joshi  3 , Ganesh V Halade  4
Affiliations

Resolvin D1 activates the inflammation resolving response at splenic and ventricular site following myocardial infarction leading to improved ventricular function

Vasundhara Kain et al. J Mol Cell Cardiol. 2015 Jul.

Abstract

Unresolved inflammation is a major contributor to the development of heart failure following myocardial infarction (MI). Pro-resolving lipid mediators, such as resolvins (e.g. RvD1), are biosynthesized endogenously. The role of RvD1 in resolving post-MI inflammation has not been elucidated due to its unstable nature. Here, we have tested the role for two forms of RvD1, after incorporation into liposomes (Lipo-RvD1) and its free acid form (RvD1) in the left ventricle (LV) and splenic remodeling post-MI. 8 to 12-week old male, C57BL/6J-mice were subjected to coronary artery ligation and Lipo-RvD1 or RvD1 (3 μg/kg/day) was injected 3h post-MI for day (d)1 or until d5. No-MI mice and saline-injected MI mice served as controls. RvD1 injected groups showed improved fractional shortening post-MI; preserving transient changes in the splenic reservoir compared to MI-saline. RvD1-groups showed an early exit of neutrophils from LV and spleen at d5 post-MI with an increased expression of lipoxin A4 receptor (ALX; synonym formyl peptide receptor; FPR2) compared to the MI-saline group. The levels of pro-resolving mediators RvD1, RvD2, Maresin 1 (MaR1) and Lipoxin A4 (LXA4) were increased in spleens from RvD1 injected mice at d5 post-MI. RvD1 administration reduced macrophage density, ccr5 and cxcl5 levels at d5 post-MI compared to saline injected mice (both, p < 0.05). Increased transcripts of mrc-1, arg-1 and Ym-1 (all, p < 0.05) suggest macrophage-mediated clearance of necrotic cells in RvD1-groups. RvD1 reduced the pro-fibrotic genes (colla1, coll2a1 and tnc (all; p < 0.05)) and decreased collagen deposition, thereby reducing post-MI fibrosis and thus stabilizing the extracellular matrix. In summary, RvD1 and Lipo-RvD1 promote the resolution of acute inflammation initiated by MI, thereby delaying the onset of heart failure.

Keywords: Metabololipidomics; Myocardial infarction; Neutrophils; Resolution of inflammation; Resolvin D1; Splenic remodeling.

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Figures

Figure 1
Figure 1. RvD1 reduced LV neutrophil density and activated ALX/FPR2 expression at d5 post-MI
(A) Representative neutrophil IHC images of LV transverse section for MI-saline, Lipo-RvD1 and RvD1 injected group at d1 and d5 post-MI (Magnification 40X, 4–6 images/slide were quantified/mouse, 3–5 mice/group, LV middle section at 1.25x, scale bar =50 µm). The arrow indicate the neutrophils at d5 post-MI in MI-saline, Lipo-RvD1 and RvD1 injected group (B) Chemical structure of RvD1, (C) Quantification of percentage stained area of neutrophils in the LV IHC at d1 and d5 post-MI in MI-saline, Lipo-RvD1 and RvD1 injected groups, the absence of quantifiable neutrophils in the LV are presented as “undetermined”. (D) mRNA expression of ALX/FPR2 in infarcted area of MI-saline, Lipo-RvD1 and RvD1 injected groups. *p<0.05 vs d0 no-MI control, $p<0.05 vs MI-saline at respective day, n=3–5 mice/group/day.
Figure 2
Figure 2. RvD1 reduced spleen neutrophils density and activated FPR2/ALX expression at d5 post-MI
(A) H& E images of the spleen of d0 controls and at d1 and d5 post-MI in MI-saline, Lipo-RvD1 and RvD1 injected group; scale bar= 50 µm, magnification=40X, 4–6 images/slide were quantified/mouse, 3–5 mice/group. (B) Quantification of the percentage stained area of neutrophils in the spleen IHC at d1 and d5 post-MI in MI-saline, Lipo-RvD1 and RvD1 injected groups. (C) mRNA expression of ALX/FPR2 in spleen in MI-saline, Lipo-RvD1 and RvD1 treated groups in control and at d1 and d5 post-MI. *p<0.05 vs d0 no-MI control, $p<0.05 vs MI-saline at respective day; n=3-5/group/day.
Figure 3
Figure 3. Mass spectrometry profiling of SPM (specialized pro-resolving lipid mediators) from DHA, EPA and arachidonic acid bioactive metabolomes in spleen at d5 post-MI
(A) Representative Multiple Reaction Monitoring (MRM) traces for the identified lipid mediators accompanying MS/MS spectra used for identification of LXA4 and MaR1. (B) Accompanying MS/MS spectra used for identification of LXA4 and MaR1.
Figure 4
Figure 4. RvD1 reduced macrophage density at d5 post-MI
(A) Representative Mac-3 IHC images of LV transverse section at d1 and d5 post-MI in MI-saline, Lipo-RvD1 and RvD1 respectively. (Magnification 40X, 4–6 images/slide were quantified/mouse, 3–5 mice/group, LV middle section at 1.25x, scale =50 µm), *p<0.05 vs MI-saline at d5 post-MI for Lipo-RvD1 and RvD1 treated groups (n=3–4 mice/group/day); (B) Macrophages (mac-3) stained area in the LV IHC at d5 post-MI in MI-saline, Lipo-RvD1 and RvD1 treated groups. (C) Venn diagram representing post-MI inflammatory gene expression in LVI regions of MI-saline, Lipo-RvD1 and RvD1 mice at d1 post-MI. (D) Venn diagram representing post-MI inflammatory gene expression in infarcted area of MI-saline, Lipo-RvD1 and RvD1 mice at d1 and d5 post-MI. Red indicates ted genes expression and green indicates led genes. Black indicates no change in genes. All the genes are normalized to no-MI control.
Figure 5
Figure 5. RvD1 promoted alternative macrophages polarization without altering pro-inflammatory response
Gene expression of (A) Classical M1 markers, (B) Alternative M2 markers in MI-saline, Lipo-RvD1 and RvD1 treated groups. *p<0.05 vs d0 control, $p<0.05 vs MI-saline at respective day; n=4 mice/group/day, Lowest gene expression values of IL-6, Arg-1 and Ym-1 at d0 control are presented as “undetermined”.
Figure 6
Figure 6. RvD1 activated 5-LOX, ALX/FPR2 and modulated COX-1 and COX-2
(A) Immunoblot representing 5-LOX expression (B) Densitometric analysis and mRNA expression of 5-LOX (C) Immunoblot representing ALX/FPR2 expression (D) Densitometric analysis and mRNA expression of ALX/FPR2 (E) Immunoblot representing COX-1 expression (F) Densitometric analysis and mRNA expression of COX-1 (G) Immunoblot representing COX-2 expression. (H) Densitometric analysis and mRNA expression of COX-2. The protein and mRNA expression is measured in LV infarct of MI-saline, Lipo-RvD1 and RvD1 injected mice at d1 and d5 post-MI. *p<0.05 vs d0 no-MI control, $p<0.05 vs MI-saline at respective day; n=3-4/group/day.
Figure 7
Figure 7. RvD1 reduced post-MI LV collagen content and extracellular matrix transcripts
(A) Representative images of PSR staining in LV infarct of MI-saline, Lipo-RvD1 and RvD1 injected mice at d1 and d5 post-MI (Magnification 40X, 4–6 images/slide were quantified/mouse, 3–5 mice/group, LV middle section at 1.25x, scale =50 µm). (B) Collagen percentage stained area in the LV at d5 post-MI in MI-saline, Lipo-RvD1 and RvD1 injected groups. *p<0.05 vs d5 MI-saline; n=3-4/group/day (C) Venn diagram representing post-MI ECM remodeling genes in LV infarct of MI-saline, Lipo-RvD1 and RvD1 injected mice at d1 post-MI. (D) Venn diagram representing post-MI ECM remodeling genes in LV infarct of MI-Saline, Lipo-RvD1 and RvD1 injected mice at d5 post-MI. Post-MI gene expression are normalized to respective d0 control. Red indicates ↑ed and green indicates ↓ed gene expression. Black indicates no change in genes.
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References

    1. Reina-Couto M, Carvalho J, Valente MJ, Vale L, Afonso J, Carvalho F, et al. Impaired resolution of inflammation in human chronic heart failure. Eur J Clin Invest. 2014;44:527–538. - PubMed
    1. Kain V, Prabhu SD, Halade GV. Inflammation revisited: inflammation versus resolution of inflammation following myocardial infarction. Basic Res Cardiol. 2014;109:444. - PubMed
    1. Serhan CN. Pro-resolving lipid mediators are leads for resolution physiology. Nature. 2014;510:92–101. - PMC - PubMed
    1. Saito T, Rodger IW, Hu F, Robinson R, Huynh T, Giaid A. Inhibition of COX pathway in experimental myocardial infarction. J Mol Cell Cardiol. 2004;37:71–77. - PubMed
    1. Kimmel SE, Berlin JA, Reilly M, Jaskowiak J, Kishel L, Chittams J, et al. Patients exposed to rofecoxib and celecoxib have different odds of nonfatal myocardial infarction. Ann Intern Med. 2005;142:157–164. - PubMed

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