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基于iTRAQ的乙型肝炎病毒所致慢加急性肝衰竭患者肝组织蛋白质组学分析

iTRAQ-based proteomic analysis of hepatic tissues from patients with hepatitis B virus-induced acute-on-chronic liver failure.

作者信息

Peng Liang, Liu Jing, Li Yang-Mei, Huang Zhan-Lian, Wang Pei-Pei, Zheng Yu-Bao, Hua Yun-Peng, Gao Zhi-Liang

机构信息

Department of Infectious Diseases, Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou, Guangdong 510630, P.R. China.

Department of Traditional Chinese Medicine, Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou, Guangdong 510630, P.R. China.

出版信息

Exp Ther Med. 2015 Nov;10(5):1732-1742. doi: 10.3892/etm.2015.2727. Epub 2015 Sep 4.

Abstract

The pathogenesis of hepatitis B virus (HBV)-induced acute-on-chronic liver failure (ACLF), a serious and prevalent medical condition, is not clear, particularly with regard to which proteins are expressed in the course of the disease. The aim of the present study was to identify the differences in hepatic tissue protein expression between normal human subjects and patients with ACLF using isobaric tags for relative and absolute quantification (iTRAQ)-based proteomic analysis and to verify the results using western blot analysis. The iTRAQ method was used to analyze the protein contents of hepatic tissue samples from 3 patients with HBV-induced ACLF and 3 normal healthy subjects. The results were verified by subjecting the hepatic tissues from 2 patients with HBV-induced ACLF and 4 healthy subjects to western blot analysis. In total, 57 proteins with ≥1.5-fold differences between patients with HBV-induced ACLF and healthy subjects were identified using iTRAQ. Among these 57 proteins, 4 with the most marked differences in their expression and the most significant association with liver disease were selected to be verified through western blot analysis: Keratin, type-I cytoskeletal 19; α-1-acid glycoprotein 1 (α1-AGP); carbonic anhydrase-1; and serpin peptidase inhibitor and clade A (α-1 anti proteinase, antitrypsin) member 1 (SERPINA1). The results of the western blot analyses were nearly identical to the iTRAQ results. Identifying the differences in liver protein expression in patients with HBV-induced ACLF may provide a basis for studies on the pathogenesis of ACLF. Future studies should focus particularly on α1-AGP, carbonic anhydrase 1 and SERPINA1.

摘要

乙型肝炎病毒(HBV)引发的慢加急性肝衰竭(ACLF)是一种严重且常见的病症,其发病机制尚不清楚,尤其是在疾病过程中哪些蛋白质会表达。本研究的目的是使用基于相对和绝对定量的等压标签(iTRAQ)蛋白质组学分析来确定正常人类受试者与ACLF患者肝组织蛋白质表达的差异,并通过蛋白质印迹分析来验证结果。采用iTRAQ方法分析了3例HBV诱导的ACLF患者和3例正常健康受试者的肝组织样本中的蛋白质含量。通过对2例HBV诱导的ACLF患者和4例健康受试者的肝组织进行蛋白质印迹分析来验证结果。使用iTRAQ共鉴定出57种在HBV诱导的ACLF患者与健康受试者之间差异≥1.5倍的蛋白质。在这57种蛋白质中,选择了4种表达差异最显著且与肝病关联最密切的蛋白质通过蛋白质印迹分析进行验证:角蛋白,I型细胞骨架19;α-1-酸性糖蛋白1(α1-AGP);碳酸酐酶-1;以及丝氨酸蛋白酶抑制剂A族(α-1抗蛋白酶,抗胰蛋白酶)成员1(SERPINA1)。蛋白质印迹分析结果与iTRAQ结果几乎相同。确定HBV诱导的ACLF患者肝脏蛋白质表达的差异可能为ACLF发病机制的研究提供依据。未来的研究应特别关注α1-AGP、碳酸酐酶1和SERPINA1。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c34/4665371/0706ff501825/etm-10-05-1732-g00.jpg

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