Hepatic ischemia/reperfusion (I/R) injury represents a major risk factor for liver transplantation and is related to graft dysfunction and acute/chronic rejection. However, a significant part of these processes remain poorly characterized. To reveal differences in the proteome during liver I/R injury, we collected human liver biopsy samples during hepatectomy before and after the Pringle maneuver and conducted a TMT-based proteomic analysis through quantitative high-throughput mass spectrometry. We used a fold-change threshold of 1.3 and a t-test p-value < 0.05 as the criteria to identify 5,257 total quantifiable proteins. The levels of 142 proteins were increased, while the levels of 103 proteins were decreased in response to hepatic I/R treatment. Bioinformatic analysis further revealed that these differentially expressed proteins are mainly involved in multiple biological functions and enzyme-regulated metabolic pathways. Most proteins whose expression was changed are related to the defense, immune and inflammatory responses as well as lipid and steroid metabolic processes. Based on this finding, we developed a panel for targeted proteomic analysis and used the parallel reaction monitoring (PRM) method, qPCR and western blotting experiments to validate alterations in the expression of some of the identified proteins. The upregulated proteins were found to be involved in immunity and inflammatory responses, and downregulated proteins were enriched in metabolic pathways. This study therefore may provide a research direction for the design of new therapeutic strategies for hepatic ischemia/reperfusion injury.

中文翻译：

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缺血/再灌注治疗后人类肝脏的整体蛋白质组分析

肝缺血/再灌注（I / R）损伤是肝移植的主要危险因素，并且与移植物功能障碍和急性/慢性排斥反应有关。但是，这些过程中有很大一部分仍缺乏良好的特性。为了揭示肝脏I / R损伤过程中蛋白质组的差异，我们在普林格尔（Pringle）手术前后在肝切除术期间收集了人类肝脏活检样品，并通过定量高通量质谱进行了基于TMT的蛋白质组学分析。我们使用1.3的倍数变化阈值和<0.05的t检验p值作为识别5257种可定量蛋白质的标准。响应肝脏I / R处理，增加了142种蛋白质的水平，而减少了103种蛋白质的水平。生物信息学分析进一步表明，这些差异表达的蛋白质主要参与多种生物学功能和酶调节的代谢途径。大多数表达改变的蛋白质与防御，免疫和炎性反应以及脂质和类固醇代谢过程有关。基于这一发现，我们开发了针对性蛋白质组分析的专家组，并使用了平行反应监测（PRM）方法，qPCR和Western blotting实验来验证某些已鉴定蛋白表达的变化。发现上调的蛋白与免疫和炎症反应有关，而下调的蛋白则富含代谢途径。