Sun Zewei, Han Jie, Zhao Wenting, Zhang Yuanyuan, Wang Shuai, Ye Lifang, Liu Tingting, Zheng Liangrong
Department of Cardiology, the First Affiliated Hospital, College of Medicine, Zhejiang University, No. 79 Qingchun Road, Hangzhou 310003, China.
Int J Mol Sci. 2014 Oct 13;15(10):18362-80. doi: 10.3390/ijms151018362.
Transient potential receptor vanilloid 1 (TRPV1) channels, which are expressed on sensory neurons, elicit cardioprotective effects during ischemia reperfusion injury by stimulating the release of neuropeptides, namely calcitonin gene-related peptide (CGRP) and substance P (SP). Recent studies show that TRPV1 channels are also expressed on cardiomyocytes and can exacerbate air pollutant-induced apoptosis. However, whether these channels present on cardiomyocytes directly modulate cell death and survival pathways during hypoxia/reoxygenation (H/R) injury remains unclear. In the present study, we investigated the role of TRPV1 in H/R induced apoptosis of H9C2 cardiomyocytes. We demonstrated that TRPV1 was indeed expressed in H9C2 cells, and activated by H/R injury. Although neuropeptide release caused by TRPV1 activation on sensory neurons elicits a cardioprotective effect, we found that capsaicin (CAP; a TRPV1 agonist) treatment of H9C2 cells paradoxically enhanced the level of apoptosis by increasing intracellular calcium and mitochondrial superoxide levels, attenuating mitochondrial membrane potential, and inhibiting mitochondrial biogenesis (measured by the expression of ATP synthase β). In contrast, treatment of cells with capsazepine (CPZ; a TRPV1 antagonist) or TRPV1 siRNA attenuated H/R induced-apoptosis. Furthermore, CAP and CPZ treatment revealed a similar effect on cell viability and mitochondrial superoxide production in primary cardiomyocytes. Finally, using both CGRP(8-37) (a CGRP receptor antagonist) and RP67580 (a SP receptor antagonist) to exclude the confounding effects of neuropeptides, we confirmed aforementioned detrimental effects as TRPV1(-/-) mouse hearts exhibited improved cardiac function during ischemia/reperfusion. In summary, direct activation of TRPV1 in myocytes exacerbates H/R-induced apoptosis, likely through calcium overload and associated mitochondrial dysfunction. Our study provides a novel understanding of the role of myocyte TRPV1 channels in ischemia/reperfusion injury that sharply contrasts with its known extracardiac neuronal effects.
瞬时受体电位香草酸受体1(TRPV1)通道表达于感觉神经元上,通过刺激神经肽(即降钙素基因相关肽(CGRP)和P物质(SP))的释放,在缺血再灌注损伤期间发挥心脏保护作用。最近的研究表明,TRPV1通道也表达于心肌细胞上,并且会加剧空气污染物诱导的细胞凋亡。然而,这些存在于心肌细胞上的通道在缺氧/复氧(H/R)损伤期间是否直接调节细胞死亡和存活途径仍不清楚。在本研究中,我们调查了TRPV1在H/R诱导的H9C2心肌细胞凋亡中的作用。我们证明TRPV1确实在H9C2细胞中表达,并被H/R损伤激活。尽管TRPV1在感觉神经元上激活引起的神经肽释放具有心脏保护作用,但我们发现辣椒素(CAP;一种TRPV1激动剂)处理H9C2细胞反而通过增加细胞内钙和线粒体超氧化物水平、减弱线粒体膜电位以及抑制线粒体生物发生(通过ATP合酶β的表达来衡量),增强了细胞凋亡水平。相反,用辣椒平(CPZ;一种TRPV1拮抗剂)或TRPV1 siRNA处理细胞可减轻H/R诱导的凋亡。此外,CAP和CPZ处理对原代心肌细胞的细胞活力和线粒体超氧化物产生显示出相似的影响。最后,使用CGRP(8 - 37)(一种CGRP受体拮抗剂)和RP67580(一种SP受体拮抗剂)来排除神经肽的混杂效应,我们证实了上述有害作用,因为TRPV1基因敲除小鼠心脏在缺血/再灌注期间心脏功能得到改善。总之,心肌细胞中TRPV1的直接激活会加剧H/R诱导的凋亡,可能是通过钙超载和相关的线粒体功能障碍。我们的研究为心肌细胞TRPV1通道在缺血/再灌注损伤中的作用提供了新的认识,这与其已知的心脏外神经元效应形成了鲜明对比。