Role of Calcium Channels in the Protective Effect of Hydrogen Sulfide in Rat Cardiomyoblasts
Role of Calcium Channels in the Protective Effect of Hydrogen Sulfide in Rat Cardiomyoblasts
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DOI：
10.1159/000358690
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发表时间：
2014-01-01
影响因子：
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通讯作者：
Mancardi, Daniele
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文献类型：
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作者：
Avanzato, Daniele;Merlino, Annalisa;Mancardi, Daniele
工作背景：硫化氢有助于减轻心肌细胞氧化应激相关损伤，但其机制尚不清楚。目的：在这里，我们研究了电压操纵性钙通道（VOCCs）作为介质的H2S对培养的大鼠心肌细胞（H9 c2）的氧化应激的有益作用。方法：采用荧光活细胞成像法测定细胞内钙信号，比色法测定细胞活力。结果如下：用H2S供体（NaHS 10 μ M）或硝苯地平（10 μ M）处理可降低静息细胞内钙浓度[Ca]，表明L型VOCCs受H2S负调节。在硝苯地平存在下，H2S仍然能够降低[Ca] i，而与硝苯地平和Ni 2（+）100 μ M共孵育完全阻止H2S依赖的[Ca] i降低，表明L型和T型VOCC均被H2S抑制。此外，在相同的实验条件下，H2S引发[Ca] i的缓慢增加，其分子性质仍有待澄清。用NaHS（10 μ M）预处理H9 c2可显著阻止H2 O2诱导的细胞死亡。这种效果是模仿预处理与L型钙通道抑制剂硝苯地平（10 μ M）。结论：这些数据提供了第一个证据表明，硫化氢保护大鼠心肌细胞对氧化的挑战，通过抑制L-型钙通道。版权所有（C）2014 S. Karger AG，巴塞尔
Background: Hydrogen sulfide contributes to the reduction of oxidative stress-related injury in cardiomyocytes but the underlying mechanism is still unclear. Aims: Here we investigated the role of voltage-operated calcium channels (VOCCs) as mediators of the beneficial effect of H2S against oxidative stress in cultured rat cardiomyoblasts (H9c2). Methods: Intracellular calcium signals were measured by fluorimetric live cell imaging and cell viability by colorimetric assay. Results: Treatment with H2S donor (NaHS 10 mu M) or Nifedipine (10 mu M) decreased resting intracellular calcium concentration [Ca] , suggesting that L-type VOCCs are negatively modulated by H2S. In the presence of Nifedipine H2S was still able to lower [Ca] i, while co-incubation with Nifedipine and Ni2(+) 100 mu M completely prevented H2S-dependent [Ca] i decrease, suggesting that both L-type and T-type VOCCs are inhibited by H2S. In addition, in the same experimental conditions, H2S triggered a slow increase of [Ca] i whose molecular nature remains to be clarified. Pretreatment of H9c2 with NaHS (10 mu M) significantly prevented cell death induced by H2O2. This effect was mimicked by pretreatment with L-Type calcium channel inhibitor Nifedipine (10 mu M). Conclusions: The data provide the first evidence that H2S protects rat cardiomyoblasts against oxidative challenge through the inhibition of L-type calcium channels. Copyright (C) 2014 S. Karger AG, Basel