β-adrenergic receptor stimulation transactivates protease-activated receptor 1 via matrix metalloproteinase 13 in cardiac cells.
β-adrenergic receptor stimulation transactivates protease-activated receptor 1 via matrix metalloproteinase 13 in cardiac cells.
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DOI：
10.1161/circulationaha.111.066787
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发表时间：
2012-06-19
期刊：
影响因子：
37.8
通讯作者：
Blaxall BC
中科院分区：
文献类型：
--
作者：
Jaffré F;Friedman AE;Hu Z;Mackman N;Blaxall BC
慢性β-肾上腺素能受体（β-AR）过度刺激是心力衰竭的标志，与心脏基质金属蛋白酶（MMPs）表达增加有关。MMP-1已显示在非心脏细胞中切割并激活蛋白酶激活受体1（PAR 1）。在此，我们假设β-AR刺激将导致心肌细胞中MMP依赖性PAR 1的反式激活。用碱性磷酸酶标记的PAR 1转导的异丙肾上腺素（ISO）刺激新生大鼠心室肌细胞（NRVM）或心脏成纤维细胞（CF）的β-AR引起AP-PAR 1切割的显著增加。广谱MMP抑制剂GM 6001显著降低了这种ISO依赖性裂解。重要的是，特异性MMP-13抑制剂还降低了ISO刺激的NRVM中的AP-PAR 1裂解，以及ISO刺激的CF中用条件培养基刺激的NRVM中的AP-PAR 1裂解。此外，我们发现重组MMP-13刺激在NRVM中的DPRS 42 ↓ 43 FLLRN处切割AP-PAR 1。NRVMs通过Gαq激活ERK 1/2通路，CFs通过Gαq/ErbBR通路激活ERK 1/2通路。MMP-13引起类似水平的ERK 1/2激活，但较低水平的肌醇磷酸生成，相比凝血酶。最后，我们证明了PAR 1基因消融或MMP-13的药理学抑制可预防小鼠ISO依赖性心功能不全。在这项研究中，我们证明β-AR刺激导致心脏成纤维细胞和心肌细胞中PAR 1的MMP-13反式激活，这可能有助于心脏中Gαq和ErbB受体依赖性途径的病理激活。我们认为这一机制可能是β-AR过度刺激依赖性心功能不全发生的基础。
Chronic β-adrenergic receptor (β-AR) overstimulation, a hallmark of heart failure, is associated with increased cardiac expression of matrix metalloproteinases (MMPs). MMP-1 has been shown to cleave and activate the protease-activated receptor 1 (PAR1) in non-cardiac cells. Here, we hypothesized that β-AR stimulation would result in MMP-dependent PAR1 transactivation in cardiac cells. β-AR stimulation of neonatal rat ventricular myocytes (NRVMs) or cardiac fibroblasts (CFs) with isoproterenol (ISO) transduced with an alkaline phosphatase-tagged PAR1 elicited a significant increase in AP-PAR1 cleavage. This ISO-dependent cleavage was significantly reduced by the broad-spectrum MMP inhibitor GM6001. Importantly, specific MMP-13 inhibitors also decreased AP-PAR1 cleavage in ISO stimulated NRVMs, as well as in NRVMs stimulated with conditioned-medium from ISO-stimulated CFs. Moreover, we found that recombinant MMP-13 stimulation cleaved AP-PAR1 in NRVMs at DPRS42↓43FLLRN. This also led to the activation of ERK1/2 pathway through Gαq in NRVMs and via the Gαq/ErbBR pathways in CFs. MMP-13 elicited similar levels of ERK1/2 activation, but lower levels of inositol phosphates generation, in comparison to thrombin. Finally, we demonstrated that either PAR1 genetic ablation or pharmacological inhibition of MMP-13 prevented ISO-dependent cardiac dysfunction in mice. In this study, we demonstrate that β-AR stimulation leads to MMP-13 transactivation of PAR1 in both cardiac fibroblasts and cardiomyocytes and this likely contributes to pathological activation of Gαq- and ErbB receptor-dependent pathways in the heart. We propose that this mechanism may underly the development of β-AR overstimulation-dependent cardiac dysfunction.