BACKGROUND: The protective role of M(3)-mAChR against apoptosis has been identified previously. However, the underlying mechanisms remain unclear. This study was performed to clarify the signaling pathways of the anti-apoptotic effect mediated by activation of M(3)-mAChR in cultured cardiac H9c2 cells. METHODS: Both H9c2 rat ventricular cells and H9c2 cells with stable expression of M(3)-mAChR were used. RESULTS: Activation of M(3)-mAChR by cabarchol produced protective effect on etoposide-induced apoptosis in H9c2 cells. Forced overexpression of M(3)-mAChR in H9c2 cells further enhanced this effect. Application of 4-diphenyl-acetoxy-N-methyl-piperidine methiodide (inhibitor of M(3)-mAChR), YC-1 [inhibitor of hypoxia-inducible factor 1, (HIF-1], or ZnPP (inhibitor of heme oxygenase-1)abrogated carbacol-induced cardioprotection, respectively. Moreover, the expression of HIF-1α, HO-1, and vascular endothelial growth factor (VEGF) were enhanced after the activation of M3-mAChR, and the induction of HO-1 and VEGF was reversed by HIF-1α inhibitor YC-1. CONCLUSIONS: These findings indicated that M(3)-mAChR upregulates HO-1 and VEGF expression likely through induction of HIF-1α, which at least partly underlies the cytoprotection of M(3)-mAChR activation in H9c2 cells.
BACKGROUND: The protective role of M(3)-mAChR against apoptosis has been identified previously. However, the underlying mechanisms remain unclear. This study was performed to clarify the signaling pathways of the anti-apoptotic effect mediated by activation of M(3)-mAChR in cultured cardiac H9c2 cells. METHODS: Both H9c2 rat ventricular cells and H9c2 cells with stable expression of M(3)-mAChR were used. RESULTS: Activation of M(3)-mAChR by cabarchol produced protective effect on etoposide-induced apoptosis in H9c2 cells. Forced overexpression of M(3)-mAChR in H9c2 cells further enhanced this effect. Application of 4-diphenyl-acetoxy-N-methyl-piperidine methiodide (inhibitor of M(3)-mAChR), YC-1 [inhibitor of hypoxia-inducible factor 1, (HIF-1], or ZnPP (inhibitor of heme oxygenase-1)abrogated carbacol-induced cardioprotection, respectively. Moreover, the expression of HIF-1α, HO-1, and vascular endothelial growth factor (VEGF) were enhanced after the activation of M3-mAChR, and the induction of HO-1 and VEGF was reversed by HIF-1α inhibitor YC-1. CONCLUSIONS: These findings indicated that M(3)-mAChR upregulates HO-1 and VEGF expression likely through induction of HIF-1α, which at least partly underlies the cytoprotection of M(3)-mAChR activation in H9c2 cells.
Authors: Panagiotis Laspas; Jan J Sniatecki; Christoph Brochhausen; Andreas Steege; Evgeny Goloborodko; Marcin L Kordasz; Franz H Grus; Norbert Pfeiffer; Adrian Gericke Journal: J Mol Neurosci Date: 2015-02-28 Impact factor: 3.444