PDF(Pubmed)
Abstract：
Mild hypothermia (MH) is an effective measure to alleviate cerebral ischemia-reperfusion (I/R) injury. However, the underlying biological mechanisms remain unclear. This study set out to investigate dynamic changes in urinary proteome due to MH in rats with cerebral I/R injury and explore the neuroprotective mechanisms of MH. A Pulsinelli\'s four-vessel occlusion (4-VO) rat model was used to mimic global cerebral I/R injury. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed to profile the urinary proteome of rats with/without MH (32 °C) treatment after I/R injury. Representative differentially expressed proteins (DEPs) associated with MH were validated by western blotting in hippocampus. A total of 597 urinary proteins were identified, among which 119 demonstrated significant changes associated with MH. Gene Ontology (GO) annotation of the DEPs revealed that MH significantly enriched in endopeptidase activity, inflammatory response, aging, response to oxidative stress and reactive oxygen species, blood coagulation, and cell adhesion. Notably, changes in 12 DEPs were significantly reversed by MH treatment. Among them, 8 differential urinary proteins were previously reported to be closely associated with brain disease, including NP, FZD1, B2M, EPCR, ATRN, MB, CA1and VPS4A. Two representative proteins (FZD1, B2M) were further validated by western blotting in the hippocampus and the results were shown to be consistent with urinary proteomic analysis. Overall, this study strengthens the idea that urinary proteome can sensitively reflect pathophysiological changes in the brain, and appears to be the first study to explore the neuroprotective effects of MH by urinary proteomic analysis. FZD1 and B2M may be involved in the most fundamental molecular biological mechanisms of MH neuroprotection.
摘要：
亚低温（MH）是减轻脑缺血再灌注（I/R）损伤的有效措施。然而,潜在的生物学机制仍不清楚.本研究旨在研究脑I/R损伤大鼠MH引起的尿蛋白质组的动态变化，并探讨MH的神经保护机制。使用Pulsinelli的四血管闭塞（4-VO）大鼠模型来模拟全脑I/R损伤。采用液相色谱-串联质谱（LC-MS/MS）对I/R损伤后接受/未接受MH（32°C）治疗的大鼠的尿液蛋白质组进行了分析。与MH相关的代表性差异表达蛋白（DEP）在海马中通过蛋白质印迹进行验证。共鉴定出597种尿蛋白,其中119例表现出与MH相关的显著变化。DEP的基因本体论(GO)注释显示MH显著富集内肽酶活性,炎症反应，老化，对氧化应激和活性氧的反应，血液凝固，和细胞粘附。值得注意的是,12个DEP的变化被MH治疗显著逆转.其中,以前报道有8种差异尿蛋白与脑部疾病密切相关，包括NP,FZD1,B2M,EPCR,ATRN,MB,CA1和VPS4A。在海马中通过蛋白质印迹进一步验证了两种代表性蛋白质(FZD1,B2M),结果显示与尿蛋白质组分析一致。总的来说,这项研究加强了尿蛋白质组可以敏感地反映大脑病理生理变化的观点，似乎是第一个通过尿蛋白质组学分析探索MH神经保护作用的研究。FZD1和B2M可能参与了MH神经保护的最基本的分子生物学机制。
