Urine Proteomic Signatures of Mild Hypothermia Treatment in Cerebral Ischemia-Reperfusion Injury in Rats.

IF 3.6 4区 医学 Q3 CELL BIOLOGY Cellular and Molecular Neurobiology Pub Date : 2024-06-05 DOI:10.1007/s10571-024-01483-4
Dandan Zhang, Dapeng Li, Xueting Wang, Yanyan Sui, Fuguo Ma, Yuting Dai, Mingshan Wang, Weiwei Qin
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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.

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轻度低温治疗大鼠脑缺血再灌注损伤的尿液蛋白质组学特征
轻度低体温(MH)是缓解脑缺血再灌注(I/R)损伤的有效措施。然而,其潜在的生物学机制仍不清楚。本研究旨在研究轻度低温导致脑缺血再灌注损伤大鼠尿蛋白质组的动态变化,并探索轻度低温的神经保护机制。研究采用普氏四血管闭塞(4-VO)大鼠模型模拟全脑I/R损伤。采用液相色谱-串联质谱(LC-MS/MS)分析了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。海马中与 MH 相关的代表性差异表达蛋白(DEPs)通过 Western 印迹进行了验证。共鉴定出 597 种尿液蛋白,其中 119 种与 MH 相关的蛋白发生了显著变化。DEPs 的基因本体(Gene Ontology,GO)注释显示,MH 在内肽酶活性、炎症反应、衰老、对氧化应激和活性氧的反应、血液凝固和细胞粘附等方面明显富集。值得注意的是,12 种 DEPs 的变化在 MH 治疗后明显逆转。其中,8种不同的尿液蛋白以前曾被报道与脑部疾病密切相关,包括NP、FZD1、B2M、EPCR、ATRN、MB、CA1和VPS4A。通过在海马中进行 Western 印迹,进一步验证了两种代表性蛋白质(FZD1、B2M),结果显示与尿液蛋白质组分析一致。总之,这项研究加强了尿液蛋白质组能灵敏反映大脑病理生理变化的观点,并且似乎是第一项通过尿液蛋白质组分析探讨 MH 神经保护作用的研究。FZD1和B2M可能参与了MH神经保护最基本的分子生物学机制。
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来源期刊
CiteScore
7.70
自引率
0.00%
发文量
137
审稿时长
4-8 weeks
期刊介绍: Cellular and Molecular Neurobiology publishes original research concerned with the analysis of neuronal and brain function at the cellular and subcellular levels. The journal offers timely, peer-reviewed articles that describe anatomic, genetic, physiologic, pharmacologic, and biochemical approaches to the study of neuronal function and the analysis of elementary mechanisms. Studies are presented on isolated mammalian tissues and intact animals, with investigations aimed at the molecular mechanisms or neuronal responses at the level of single cells. Cellular and Molecular Neurobiology also presents studies of the effects of neurons on other organ systems, such as analysis of the electrical or biochemical response to neurotransmitters or neurohormones on smooth muscle or gland cells.
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