Abstract 186: AMP-Activated Protein Kinase Mediates Cardioprotection in the Regenerative MRL Mouse Strain

Abstract

Introduction: The Murphy Roths Large (MRL/MpJ) mouse model is well-known for its healing and regenerative properties. The actual mechanisms promoting regeneration are still unknown. AMP activated protein kinase (AMPK) has been lauded for its role as a cardioprotective sensor of cellular metabolic stress, cell growth, and ATP depletion. Based on our preliminary data, we hypothesize that increased levels of AMPK in the MRL hearts is primarily responsible for its altered mitochondrial biogenesis and metabolism, reduced oxidative stress, and altered cardiac function in the MRL mice. Methods: Adult C57BL/6 (control) and MRL mice (n=5/group) were studied pre- and post-7 days of exercise, and post-exercise with 3 days injection of either 5-amino -imidazole carboxamide riboside (AICAR, AMPK stimulator, 250ug/g, IP), or Compound C (AMPK inhibitor, 20ug/g, IP). Results: Compared to control at baseline, the MRL hearts had higher AMPK protein levels (9 fold increase, p<0.001), equivalent AMPK mRNA levels by qRT-PCR, and higher AMP to ATP ratios by HPLC (3 fold increase, p>0.05, n.s.). The MRL hearts also had increased mitochondria by electron microscopy (by 65 ± 4.5%, p<0.05), reduced HW to BW ratio (48% decrease, p<0.001), reduced cellular and mitochondrial reactive oxygen species (by DHE and Mitosox stains respectively), and increased glycolysis (Glut-4 sarcolemmal translocation). Hypertrophic response to exercise was greater in the MRL mice (HW/BW increased by 40% vs. 2% in control, p<0.001). Mimicking injury, Compound C reduced active phosphorylated AMPK protein levels in control (by 26%, p<0.05), while increased it in MRL mice (by 30%, p<0.05). AICAR injection significantly increased pAMPK protein in both strains (p=0.008). Baseline MRL mice echocardiographic analysis (vs. control) indicated higher systolic function, but increased diastolic E/A and E/E’ ratios (p<0.05). In the MRL hearts, Compound C treatment reduced %EF and %FS, while AICAR reduced post-exercise hypertrophy (p=0.02). Conclusions: This study highlights the cardioprotective role AMPK plays in the MRL mice, through its modulation of mitochondrial function and reduction of oxidative stress. This can in turn be used to develop new therapeutic treatment for heart failure.