Transient receptor potential melastatin 4 inhibition prevents lipopolysaccharide-induced endothelial cell death
Transient receptor potential melastatin 4 inhibition prevents lipopolysaccharide-induced endothelial cell death
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DOI：
10.1093/cvr/cvr135
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发表时间：
2011-09-01
影响因子：
10.8
通讯作者：
Simon, Felipe
中科院分区：
文献类型：
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作者：
Becerra, Alvaro;Echeverria, Cesar;Simon, Felipe
目的内皮功能障碍在心血管疾病的进展中起决定性作用。脂多糖（LPS）诱导的活性氧（ROS）介导的内皮细胞死亡是继发于内毒素血症的炎症中观察到的主要特征，是重症监护室危重患者死亡的主要原因。然而，LPS诱导的内皮细胞死亡的分子机制还不清楚。瞬时受体蛋白melastatin 4（TRPM 4）是一种与细胞死亡相关的离子通道，在内皮细胞中表达并受ROS调节。在这里，我们调查TRPM 4在LPS诱导的内皮细胞死亡的作用，测试是否抑制TRPM 4的表达赋予内皮细胞抵抗LPS challenge.Methods和结果使用原代培养人脐静脉内皮细胞（HUVEC），我们证明，TRPM 4是关键参与LPS诱导的内皮细胞死亡。暴露于LPS的HUVEC导致Na+依赖性细胞死亡。TRPM 4与9-菲咯或格列本脲的药理学抑制作用可保护内皮免受LPS暴露48小时的影响。此外，TRPM 4样电流在LPS预处理的细胞中增加，并被格列本脲抑制。值得注意的是，抑制TRPM 4表达的siRNA或抑制其活性的显性负突变体是有效的减少LPS诱导的内皮细胞死亡时，细胞暴露于LPS为24-30小时。结论TRPM 4是关键参与LPS诱导的内皮细胞死亡。这些结果表明，TRPM 4的药理学抑制、TRPM 4表达的抑制或TRPM 4活性的抑制能够保护内皮免受LPS损伤。这些结果有助于脓毒症药物的设计和脓毒症治疗新策略的开发。
Aims Endothelial dysfunction is decisive in the progression of cardiovascular diseases. Lipopolysaccharide (LPS)-induced reactive oxygen species (ROS)-mediated endothelial cell death is a main feature observed in inflammation secondary to endotoxaemia, emerging as a leading cause of death among critically ill patients in intensive care units. However, the molecular mechanism underlying LPS-induced endothelial cell death is not well understood. Transient receptor protein melastatin 4 (TRPM4) is an ion channel associated with cell death that is expressed in endothelium and modulated by ROS. Here, we investigate the role of TRPM4 in LPS-induced endothelial cell death, testing whether suppression of the expression of TRPM4 confers endothelial cell resistance to LPS challenge.Methods and results Using primary cultures of human umbilical vein endothelial cells (HUVEC), we demonstrate that TRPM4 is critically involved in LPS-induced endothelial cell death. HUVEC exposed to LPS results in Na+-dependent cell death. Pharmacological inhibition of TRPM4 with 9-phenanthrol or glibenclamide protects endothelium against LPS exposure for 48 h. Furthermore, TRPM4-like currents increase in cells pre-treated with LPS and inhibited with glibenclamide. Of note, suppression of TRPM4 expression by siRNA or suppression of its activity in a dominant negative mutant is effective in decreasing LPS-induced endothelial cell death when cells are exposed to LPS for 24-30 h.Conclusion TRPM4 is critically involved in LPS-induced endothelial cell death. These results demonstrate that either pharmacological inhibition of TRPM4, suppression of TRPM4 expression, or inhibition of TRPM4 activity are able to protect endothelium against LPS injury. These results are useful in sepsis drug design and development of new strategies for sepsis therapy.