基于蛋白质组学方法探讨四氯化碳诱导的小鼠急性肝损伤的差异蛋白表达  

Exploring the Relative Differential Protein Expression of Carbon Tetrachloride-induced Acute Liver Injury in Mice Based on the Proteomics Method

作　　者：黎伟华 吴璟 金学琴 雷艳丽 LI Wei-hua;WU Jing;JIN Xue-qin;LEI Yan-li(College of Basic Medicine,Ningxia Medical University,Yinchuan 750004;College of Pharmacy,Ningxia Medical University,Yinchuan 750004)

机构地区：[1]宁夏医科大学基础医学院,银川750004 [2]宁夏医科大学药学院,银川750004

出　　处：《生物技术通报》2025年第2期331-342,共12页Biotechnology Bulletin

基　　金：宁夏自然科学基金项目(2022AAC03202)。

摘　　要：【目的】应用标记定量(TMT)蛋白质组学技术,探究化学诱导剂(carbon tetrachloride,CCl4)致小鼠急性肝损伤的作用机制。【方法】20只SPF级雄性BALB/C小鼠随机分为对照组和诱导组,每组10只,腹腔注射诱导8周(2次/周)。通过测定血液生化水平及肝组织病理切片等指标评价肝损伤的程度。通过串联质谱标记技术(TMT)进行蛋白质组学分析,对发现的差异蛋白进行基因本体论(GO)注释、KEGG通路和蛋白互作网络分析。【结果】与对照组比较,CCl4诱导组的血液生化指标谷草转氨酶(aspartate aminotransferase,AST)的值显著升高(P<0.05);谷丙转氨酶(alanine aminotransferase,ALT)、谷氨酰转肽酶(Yglutamyl transpeptadase,GGT)、总胆汁酸(total biliary acid,TBA)的值极显著增加(P<0.01),且肝组织病理切片显示CCl4诱导组小鼠肝损伤明显。利用TMT技术共鉴定了5955个蛋白,其中440个蛋白强度增加,294个蛋白强度降低,差异蛋白在蛋白表达差异倍数、生物标志物应用和参与经典信号通路(JAK-STAT、PI3K-Akt、Rap1、GnRH、AGE-RAGE)中发挥重要作用。经典信号通路中差异蛋白存在显著富集,且富集程度相对较高。【结论】CCl_(4)作为肝损伤评价的理想化学诱导剂,参与了机体免疫、细胞分裂、凋亡和自噬、炎症和肿瘤形成等多个关键过程,多方向多靶点还原了肝损伤的病理机制,为保肝药物的研发提供了坚实的理论基础和视角。【Objective】Labker quantitative(TMT)proteomics was used to explore the mechanism of chemical inducer(carbon tetrachloride,CCl4)causing acute liver injury in mice.【Method】Twenty SPF male BALB/C mice were randomly divided into control and induction groups,with 10 mice in each group,induced by intraperitoneal injection for 8 weeks(2 times/week).The degree of liver injury was evaluated by measuring the blood biochemical level and liver histopathological sections.Proteomic analysis was performed by tandem mass spectrometry labeling technology(TMT),including gene Ontology(GO)annotation,KEGG pathways,and protein interaction network analysis of the found differential proteins.【Result】Compared with the control group,blood indicator of aspartate aminotransferase(AST)was significantly higher(P<0.05);alanine aminotransferase(ALT),y-glutamyl transpeptadase(GGT),total biliary acid(TBA)(P<0.01).Moreover,liver histopathological sections showed significant liver injury in the CCl4 induction group.A total of 5955 proteins were identified using TMT,with 440 increased intensity and 294 decreased,and differential proteins played important roles in protein expression difference,biomarker application and participation in classical signaling pathways(JAK-STAT,PI3K-Akt,Rap 1,GnRH,AGE-RAGE).There was a significant enrichment of differential proteins in classical signaling pathways and a relatively high enrichment.【Conclusion】CCl_(4) as an ideal chemical inducer for liver injury evaluation,it is involved in several key processes such as immunity,cell division,apoptosis and autophagy,inflammation and tumor formation.It reduces the pathological mechanism of liver injury in multiple directions and targets and provides a solid theoretical basis and perspective for the research and development of liver protection drugs.

关 键 词：蛋白质组学 四氯化碳 肝损伤 小鼠 

分 类 号：R73[医药卫生—肿瘤]