Role of the TRPM4 channel in mitochondrial function, calcium release, and ROS generation in oxidative stress
Role of the TRPM4 channel in mitochondrial function, calcium release, and ROS generation in oxidative stress
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TRPM4 通道在氧化应激中线粒体功能、钙释放和 ROS 生成中的作用
DOI：
10.1016/j.bbrc.2021.03.077
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发表时间：
2021
影响因子：
3.1
通讯作者：
Takahashi Ken
中科院分区：
文献类型：
--
作者：
Wang Chen;Chen Jian;Wang Mengxue;Naruse Keiji;Takahashi Ken
缺血性心脏病是全球最常见的死亡原因之一。线粒体功能障碍、活性氧（reactive oxygen species，ROS）产生过多和钙超载是导致心肌缺血再灌注（ischemia-reperfusion，I/R）损伤后死亡的三个关键因素。TRPM 4是一种钙激活的非选择性阳离子通道，抑制TRPM 4可保护大鼠心脏免受I/R损伤，但这种作用的具体机制尚不清楚。在这项研究中，我们研究了通过TRPM 4对I/R损伤的心脏保护机制，使用过氧化氢（H2 O2），氧化应激的主要贡献者，作为I/R损伤模型。我们使用CRISPR/Cas9敲除了大鼠心肌细胞系H9 c2中的TRPM 4基因。H2 O2处理后，野生型（WT）细胞内Ca 2+水平和ROS产生增加，而TRPM 4敲除（TRPM 4KO）细胞内Ca 2+水平和ROS产生增加。通过这种处理，线粒体功能的两个指标，线粒体膜电位（Δ Km）和细胞内ATP水平，在WT中降低，但在TRPM 4KO细胞中没有降低。总之，这些发现表明，阻断TRPM 4通道可能通过维持线粒体膜电位和细胞内ATP水平，可能通过防止细胞内Ca 2+和ROS的异常增加，保护心肌免受氧化应激。
Ischemic heart disease is one of the most common causes of death worldwide. Mitochondrial dysfunction, excessive reactive oxygen species (ROS) generation, and calcium (Ca2+) overload are three key factors leading to myocardial death during ischemia-reperfusion (I/R) injury. Inhibition of TRPM4, a Ca2+-activated nonselective cation channel, protects the rat heart from I/R injury, but the specific mechanism underlying this effect is unclear. In this study, we investigated the mechanism of cardioprotection against I/R injury via TRPM4 using hydrogen peroxide (H2O2), a major contributor to oxidative stress, as an I/R injury model. We knocked out theTRPM4gene in the rat cardiomyocyte cell line H9c2 using CRISPR/Cas9. Upon H2O2treatment, intracellular Ca2+level and ROS production increased in wild type (WT) cells but not in TRPM4 knockout (TRPM4KO) cells. With this treatment, two indicators of mitochondrial function, mitochondrial membrane potential (ΔΨm) and intracellular ATP levels, decreased in WT but not in TRPM4KOcells. Taken together, these findings suggest that blockade of the TRPM4 channel might protect the myocardium from oxidative stress by maintaining the mitochondrial membrane potential and intracellular ATP levels, possibly through preventing aberrant increases in intracellular Ca2+and ROS.