Department of Anesthesiology, Qingdao Municipal Hospital, Qingdao University, Qingdao, 266071, China.
Department of Anesthesiology, Qingdao Municipal Hospital, Dalian Medical University, Dalian, 116044, China.
J Mol Neurosci. 2022 Sep;72(9):2020-2029. doi: 10.1007/s12031-022-02055-1. Epub 2022 Aug 3.
Cerebral ischemia-reperfusion (I/R) injury is the leading cause of death in severe hypotension caused by cardiac arrest, drowning, and excessive blood loss. Urine can sensitively reflect pathophysiological changes in the brain even at an early stage. In this study, a rat model of global cerebral I/R injury was established via Pulsinelli's four-vessel occlusion (4-VO) method. Overall, 164 urinary proteins significantly changed in the 4-VO rat urine samples compared to the control samples by data-independent acquisition (DIA) proteomics technique (1.5-fold change, p < 0.05). Gene Ontology annotation showed that the acute-phase response, the ERK1 and ERK2 cascade, endopeptidase activity, blood coagulation, and angiogenesis were overrepresented. After parallel reaction monitoring (PRM) validation, 15 differential proteins having human orthologs were verified as the potential urinary markers associated with cerebral I/R injury. Of these potential biomarkers, 8 proteins were reported to be closely associated with cerebral I/R injury. Nine differential proteins changed even when there were no clinical manifestations or histopathological cerebral damage, including FGG, COMP, TFF2, HG2A, KNG1, CATZ, PTGDS, PRVA, and HEPC. These 9 proteins are potential biomarkers for early screening of cerebral I/R injury to prevent the development of cerebral injury. KNG1, CATZ, PTGDS, PRVA, and HEPC showed an overall trend of upregulation or downregulation at 12 and 48 h after I/R injury, reflecting the progression of cerebral I/R injury. These 5 proteins may serve as potential biomarkers for prognostic evaluation of cerebral I/R injury. These findings provide important clues to inform the monitoring of cerebral I/R injury and further the current understanding of its molecular biological mechanisms.
脑缺血再灌注（I/R）损伤是心脏骤停、溺水和大量失血导致严重低血压的主要死亡原因。尿液甚至在早期阶段就能敏感地反映脑的病理生理变化。在这项研究中，通过 Pulsinelli 的四血管闭塞（4-VO）方法建立了全脑 I/R 损伤大鼠模型。通过无标记定量（DIA）蛋白质组学技术，与对照样本相比，4-VO 大鼠尿液样本中有 164 种蛋白质显著变化（1.5 倍变化，p<0.05）。基因本体论注释显示，急性期反应、ERK1 和 ERK2 级联、内肽酶活性、凝血和血管生成过度表达。经过平行反应监测（PRM）验证，有 15 种具有人类同源物的差异蛋白被验证为与脑 I/R 损伤相关的潜在尿标志物。在这些潜在的生物标志物中，有 8 种蛋白被报道与脑 I/R 损伤密切相关。即使在没有临床表现或组织病理学脑损伤的情况下，也有 9 种差异蛋白发生变化，包括 FGG、COMP、TFF2、HG2A、KNG1、CATZ、PTGDS、PRVA 和 HEPC。这 9 种蛋白是早期筛选脑 I/R 损伤以预防脑损伤的潜在生物标志物。KNG1、CATZ、PTGDS、PRVA 和 HEPC 在 I/R 损伤后 12 和 48 小时呈整体上调或下调趋势，反映了脑 I/R 损伤的进展。这 5 种蛋白可能作为脑 I/R 损伤预后评估的潜在生物标志物。这些发现为脑 I/R 损伤的监测提供了重要线索，并进一步加深了对其分子生物学机制的理解。
