Relaxin Antagonizes Hypertrophy and Apoptosis in Neonatal Rat Cardiomyocytes

Abstract

The pregnancy hormone relaxin has recently been shown to be cardio-protective. Despite its well-established antifibrotic actions in the heart, the effects of relaxin on cardiomyocytes (CM) remain to be determined. We investigated effects of isoform 2 of the human relaxin (H2-relaxin) on CM hypertrophy and apoptosis. In cultured neonatal rat CM, phenylephrine (50 microM) and cardiac fibroblast-conditioned medium were used respectively to induce CM hypertrophy. The degree of hypertrophy was indicated by increased cell size, protein synthesis and gene expression of atrial natriuretic peptide. Although H2-relaxin (16.7 nM) alone failed to suppress hypertrophy induced by phenylephrine, it repressed the cardiac fibroblast-conditioned medium-induced increase in protein synthesis by 24% (P<0.05) and reversed the increase in cell size (P<0.001) and atrial natriuretic peptide expression (P<0.01). We further studied the effect of H2-relaxin on CM apoptosis induced by H2O2 (200 microM). Studies of DNA laddering and nuclear staining demonstrated that H2-relaxin treatment reduced H2O2-induced DNA fragmentation. Real-time PCR and Western blot analysis revealed a significant increase in the Bcl2/Bax ratio in H2-relaxin-treated CM. Further analysis showed that activation of Akt (1.8-fold, P<0.001) and ERK (2.0-fold, P<0.01) were involved in the antiapoptotic action of H2-relaxin in CM, and that Gi/o coupling of relaxin receptors was associated with the H2-relaxin-induced Akt activation in CM. In conclusion, these results extend our current knowledge of the cardiac actions of relaxin by demonstrating that H2-relaxin indirectly inhibits CM hypertrophy and directly protects CM from apoptosis.