MCP-1 mediates ischemia/reperfusion-induced cardiomyocyte apoptosis via MCPIP1 and CaSR.

Monocyte chemotactic protein-1 (MCP-1) plays a crucial role in ischemia-reperfusion (I/R) injury; however, the detailed mechanism of MCP-1 in I/R injury-induced cardiomyocyte apoptosis remains unclear. In this study, we explored the cascade downstream of I/R-induced MCP-1 that modulates cell apoptosis and determined whether Ca2+-sensing receptors (CaSRs) are involved in the process. Protein levels were detected in a cardiac muscle cell line (HL-1) and primary cultured neonatal mouse ventricular cardiomyocytes using Western blotting and immunocytochemistry. Released MCP-1 was detected using ELISA. Both Hoechst staining and flow cytometry methods were used to measure cell apoptosis. Specific pharmacological inhibitors of CC chemokine receptor 2 (RS-102895) and CaSR (NPS-2143) as well as a CaSR activator (evocalcet) were applied to confirm the roles of these factors in I/R-induced cell apoptosis. I/R inhibited cell viability and upregulated cell apoptosis. Moreover, I/R induced the release of MCP-1 from both HL-1 cells and primary cardiomyocytes. Further research confirmed that CaSR acted as an upstream effector of monocyte chemotactic protein-1-induced protein-1 (MCPIP1) and coordinately regulated cell apoptosis, which was verified by addition of an inhibitor or activator of CaSR. Moreover, MCPIP1 induced cell apoptosis through endoplasmic reticulum (ER) stress but not autophagy induced by I/R. Based on these findings, I/R-induced MCP-1 release regulates cardiomyocyte apoptosis via the MCPIP1 and CaSR pathways, suggesting a new therapeutic strategy for I/R injury. NEW & NOTEWORTHY Ischemia-reperfusion (I/R)-induced monocyte chemotactic protein-1 release regulates cardiomyocyte apoptosis via the monocyte chemotactic protein-1-induced protein-1 (MCPIP1) and Ca2+-sensing receptor pathway. The functional changes mediated by MCPIP1 involve the activation of endoplasmic reticulum stress, but not the autophagy pathway, after I/R injury.