Proteomic analysis of extracellular vesicles released from heat-stroked hepatocytes reveals promotion of programmed cell death pathway
Proteomic analysis of extracellular vesicles released from heat-stroked hepatocytes reveals promotion of programmed cell death pathway
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对中暑肝细胞释放的细胞外囊泡的蛋白质组学分析揭示了程序性细胞死亡途径的促进
DOI：
10.1016/j.biopha.2020.110489
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发表时间：
2020-09-01
影响因子：
7.5
通讯作者：
Li, Xu
中科院分区：
文献类型：
--
作者：
Li, Yue;Zhu, Xintao;Li, Xu
肝损伤是严重中暑（HS）的常见并发症。细胞外囊泡（EV）是介导细胞间通讯的新途径的一部分。EV是否参与HS诱导的肝损伤的发病机制仍不清楚。在此，我们探讨了肝细胞EV在HS诱导的肝损伤中的作用及其在HS诱导后的蛋白调节模式。同位素标记相对和绝对定量技术（iTRAQ）和液相色谱-串联质谱分析确定了肝细胞来源的热中风EV的蛋白质组学特征的变化，我们确定了53个上调蛋白质。生物信息学分析证实，程序性细胞死亡的调节是最重要的改变途径。为了阐明HS肝细胞衍生的EV在诱导肝细胞程序性死亡和损伤中的作用，将它们添加到受体肝细胞中并注射到小鼠中。这种处理显著诱导了细胞凋亡的合成（caspase-3/8）和坏死性凋亡相关蛋白[受体相互作用蛋白1（RIP 1）、RIP 3和混合谱系激酶结构域样蛋白];肝细胞和肝组织中凋亡和坏死细胞数量增加，肝损伤生化标志物水平升高（丙氨酸转氨酶、天冬氨酸转氨酶和乳酸脱氢酶）。我们的研究是第一次全面分析肝细胞来源的热休克EV蛋白质组确认诱导肝损伤的EV。我们提供了一个新的解释HS诱导的肝损伤的病理机制。
Liver injury is a common complication of severe heat stroke (HS). Extracellular vesicles (EVs) are part of a novel pathway mediating intercellular communication. Whether EVs are involved in the pathogenesis underlying HS-induced liver injury remains unknown. Here, we explored the role of hepatocyte EVs in HS-induced liver injury and their protein regulation patterns after HS induction. Isobaric tags for relative and absolute quantification technology (iTRAQ) and liquid chromatography-tandem mass spectrometry analysis identified changes in the proteomic profiles of hepatocyte-derived heat-stroked EVs, and we identified 53 up-regulated proteins. Bioinformatics analysis verified that the regulation of programmed cell death was the most significant altered pathway. To clarify the effect of HS hepatocyte-derived EVs in inducing hepatocyte-programmed death and injury, they were added to recipient hepatocytes and injected into mice. This treatment significantly induced the synthesis of apoptosis (caspase-3/8) and necroptosis-associated proteins [receptor-interacting protein 1 (RIP1), RIP3, and mixed lineage kinase domain-like protein]; moreover, it increased the numbers of apoptotic and necroptotic cells in hepatocytes and liver tissues and increased the levels of biochemical liver injury markers (alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase). Our study is the first comprehensive analysis of the hepatocyte-derived heat-stroked EV proteome confirming the induction of liver injury by Evs. We provide a novel explanation for the pathological mechanism underlying HS-induced liver injury.