Research On Serum Differential Proteomics Of Patients With Hepatic Echinococcosis Based On DIA Technique

Objective: to screen differential proteins in serum of patients with AE(Alveolar Echinocococosis)、 CE(Cystic Echinococcosis)and CON(Control group),by DIA proteomics,and to analyze the biological process,cell composition,molecular function and signal pathway involved in differentially expressed proteins by bioinformatics analysis.The specific markers of serum proteomics of hepatic echinococcosis were preliminarily screened to provide a basis for the diagnosis,molecular mechanism and therapeutic targets of hepatic echinococcosis.Methods: 10 patients with AE,CE and 10 healthy outpatients were collected from Qinghai Provincial people’s Hospital from June 2019 to October 2019.The total protein in serum was extracted,some of which were determined by protein concentration determination and SDS-PAGE detection,and the other part was hydrolyzed by trypsin.After desalination of the hydrolyzed peptide,the sample was identified by LC-MS/MS.LC-MS/MS identification is divided into two main steps.In the first step,the traditional DDA method is used to establish the protein map database.In the second step,the DIA technology is used to collect the mass spectrum data of each sample,and the spectrum matching,quantitative information extraction and subsequent statistical analysis are carried out on the basis of DDA database.Trusted proteins and differential proteins were screened by data independent acquisition(Data independent acquisition,DIA)technology,and then the functional analysis of differential proteins was carried out,including GO analysis,KEGGPathway analysis,protein interaction analysis and correlation analysis.Combined with the differential expression of proteins and the results of bioinformation analysis,the potential biomarkers were preliminarily screened out.According to the data,key proteins and their functions and pathways were selected for follow-up key research and verification direction.Results:(1)A total of 402 proteins and 7133 peptides were identified in this DIA test,and 210 differential proteins were screened,of which 99 differential proteins were screened between the AE group and the control group,of which 41 proteins were up-regulated and 58 proteins were down-regulated.56 differential proteins were screened between the CE group and the control group,including 27 up-regulated proteins and 29 down-regulated proteins.55 differential proteins were screened between AE group and CE group,including 25 up-regulated proteins and 30down-regulated proteins.(2)the results of GO analysis showed that there were 23 biological processes,20 cell components and 21 molecular functions in AE-CON group,25 biological processes,9 cell components and 8 molecular functions in CE-CON group,and 25 biological processes,10 cell components and 8 molecular functions in ICR group DEPs.Through venn analysis of multi-group data,it was found that one kind of differential protein appeared in the three groups at the same time,38 kinds of differential protein only existed in AE group and control group,13 kinds of differential protein existed only in CE group and control group,and 16 kinds of differential protein existed in AE group and CE group.Conclusion: DIA can screen the differential proteins in the serum of patients with hepatic echinococcosis,provide a research basis for exploring the molecular mechanism of hepatic echinococcosis,and explore a new method for early diagnosis of hepatic echinococcosis.