Zhu Y, Zhang S, Yang C, Xue W, Zhang J, Li J, Zhao J, Xu J, Huang W
Department of Infectious Diseases, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2021 Aug 31;41(9):1381-1387. doi: 10.12122/j.issn.1673-4254.2021.09.13.
To screen differentially expressed proteins (DSPs) in the liver tissues of patients with nonalcoholic steatohepatitis (NASH) using proteomic technologies to identify potential therapeutic targets of NASH.
Liver tissue specimens were obtained from 3 patients with pathologically confirmed NASH and 3 normal control subjects. The total proteins were extracted from the specimens, and iTRAQ reagent was used to label the peptides for liquid chromatography tandem mass spectrometry (LC-MS/MS) detection. The DSPs were identified by comparing the data against UniProt protein database using Mascot2.3.02 software and were annotated and enriched using GO database; KEGG database was used for enrichment of the pathways involving these proteins. Real-time fluorescent quantitative PCR (qPCR) was performed to detect the mRNA expressions of the significant DSPs in NASH.
By the criteria that a DSP has >1.2 or < 0.8 fold difference between NASH group and the control group and with < 0.05 as the threshold, a total of 648 significant DSPs in NASH were identified, including 246 up-regulated and 402 down-regulated proteins. GO functional enrichment analysis showed that the DSPs were involved mainly in small molecule metabolism, organic acid metabolism, oxygen acid metabolism and other biological processes, and were enriched in KEGG pathways including the metabolic pathways, complement coagulation cascades, and ribosomes. Among the 25 DEPs with a fold difference >2.0 or < 0.5 ( < 0.05), 6 proteins showed consistent results between qPCR verification and proteomic analysis, including 5 down-regulated proteins: Jumonji protein (JARID2), Lebasillinlike protein (LCA5L), synaptophysin 1 (SYN1) and collagen α-1 (XIII) chain (COL13A1), FYVE, RhoGEF and PH domain protein 5 (FGD5), and 1 upregulated protein glutathione S-transferase Mu 4 (GSTM4).
We identified 648 DEPs inthe liver tissue of patients NASH using iTRAQ technology and bioinformatics methods, and among them JARID2, SYN1, COL13A1, FGD5, and GSTM4 may serve as the key target proteins of NASH.
利用蛋白质组学技术筛选非酒精性脂肪性肝炎（NASH）患者肝组织中的差异表达蛋白（DSPs），以确定NASH潜在的治疗靶点。
从3例经病理证实的NASH患者和3例正常对照者获取肝组织标本。从标本中提取总蛋白，并用iTRAQ试剂标记肽段用于液相色谱串联质谱（LC-MS/MS）检测。通过使用Mascot2.3.02软件将数据与UniProt蛋白质数据库进行比对来鉴定DSPs，并使用GO数据库进行注释和富集；KEGG数据库用于富集涉及这些蛋白质的通路。进行实时荧光定量PCR（qPCR）以检测NASH中显著DSPs的mRNA表达。
以DSP在NASH组与对照组之间差异倍数>1.2或<0.8且以<0.05为阈值的标准，共鉴定出NASH中648个显著的DSPs，包括246个上调蛋白和402个下调蛋白。GO功能富集分析表明，DSPs主要参与小分子代谢、有机酸代谢、含氧酸代谢等生物学过程，并在KEGG通路中富集，包括代谢通路、补体凝血级联反应和核糖体。在差异倍数>2.0或<0.5（<0.05）的25个差异表达蛋白（DEPs）中，6种蛋白质在qPCR验证和蛋白质组学分析之间显示出一致的结果，包括5种下调蛋白：Jumonji蛋白（JARID2）、Lebasillin样蛋白（LCA5L）、突触素1（SYN1）和胶原蛋白α-1（XIII）链（COL13A1）、FYVE、RhoGEF和PH结构域蛋白5（FGD5），以及1种上调蛋白谷胱甘肽S-转移酶Mu 4（GSTM4）。
我们使用iTRAQ技术和生物信息学方法在NASH患者肝组织中鉴定出648个DEPs，其中JARID2、SYN1、COL13A1、FGD5和GSTM4可能作为NASH的关键靶蛋白。
