Multi-Analyte Profiling Reveals Matrix Metalloproteinase-9 and Monocyte Chemotactic Protein-1 as Plasma Biomarkers of Cardiac Aging

Abstract

Background— We have previously shown that cardiac sarcopenia occurs with age in C57/BL6J mice. However, underlying mechanisms and plasma biomarkers of cardiac aging have not been identified. Accordingly, the objective of this study was to identify and evaluate plasma biomarkers that reflect cardiac aging phenotypes. Methods and Results— Plasma from adult (7.5±0.5 months old, n=27) and senescent (31.7±0.5 months old, n=25) C57/BL6J mice was collected, and levels of 69 markers were measured by multi-analyte profiling. Of these, 26 analytes were significantly increased and 3 were significantly decreased in the senescent group compared with the adult group. The majority of analytes that increased in the senescent group were inflammatory markers associated with macrophage functions, including matrix metalloproteinase-9 (MMP-9) and monocyte chemotactic protein-1 (MCP-1/CCL-2). Immunoblotting (n=12/group) showed higher MMP-9 and MCP-1 levels in the left ventricle (LV) of senescent mice ( P <0.05), and their expression levels in the LV correlated with plasma levels (ρ=0.50 for MMP-9 and ρ =0.62 for MCP1, P <0.05). Further, increased plasma MCP-1 and MMP-9 levels correlated with the increase in end-diastolic dimensions that occurs with senescence. Immunohistochemistry (n=3/group) for Mac-3, a macrophage marker, showed increased macrophage densities in the senescent LV, and dual-labeling immunohistochemistry of Mac-3 and MMP-9 revealed robust colocalization of MMP-9 to the macrophages in the senescent LV sections, indicating that the macrophage is a major contributor of MMP-9 in the senescent LV. Conclusions— Our results suggest that MCP-1 and MMP-9 are potential plasma markers for cardiac aging and that augmented MCP-1 and MMP-9 levels and macrophage content in the LV could provide an underlying inflammatory mechanism of cardiac aging.