Screening And Validation Of Genes Differentially Expressed In Peripheral Blood Mononuclear Cells Of Maintenance Hemodialysis Patients

Objectives:The aim of this study was to analyze the changes of m RNA expression profiles in peripheral blood mononuclear cells(PBMC)of maintenance hemodialysis(MHD)patients and healthy people based on RNA sequencing(RNA SEQ),and to screen the corresponding differentially expressed genes(DEGs)by enrichment analysis,Objective to explore the potential key genes and biological signal pathways in the occurrence and development of end-stage renal disease(ESRD),and to provide theoretical basis for the molecular mechanism,diagnosis and treatment of ESRD.Methods:Ten ESRD patients who were MHD and qualified for inclusion were selected as experimental group from May 2020 to June 2020,and 12 healthy control groups were selected to collect PBMC samples respectively.The difference of genes was analyzed by edger,and the significance of go and KEGG was analyzed for DEGs.Then,the protein protein interaction(PPI)network was constructed by string online platform,and the visualization was carried out by the software of cyberscape.The key molecular modules were constructed by using mcode plug-in,and the differential genes in the modules were enriched and analyzed to obtain the biological functions represented by each module,and the key genes were screened by the cytohubba plug-in.Finally,the quantitative real time PCR(qPCR)was used to verify 33 clinical samples,and the receiver operating characteristic(ROC)curve was drawn to evaluate the diagnostic value of three key genes combined prediction for ESRD.Results:(1)There were significant differences in gene expression profiles between the two groups.A total of 528 DEGs were screened.Compared with the healthy group,289 genes were up-regulated and 239 genes were down regulated in MHD group.(2)DEGs mainly focus on oxidative stress,immune response,inflammation,fibrosis and other biological functions and signaling pathways,and participate in the occurrence and development of ESRD.(3)The analysis of key molecular modules showed that the modules with the highest score were mainly related to biological functions or structures such as immune response,proteolysis,redox reaction,inflammatory response and extracellular matrix.(4)The three core genes CHI3L1、PGLYRP-1、CEACAM8 may be the potential key genes in the occurrence and development of ESRD.Further verification by qPCR showed that the expression levels of these three genes in MHD patients were significantly higher than those in healthy group,and the difference was statistically significant(P < 0.05).(5)The area under ROC curve of combined prediction was 0.859(95% CI: 0.7269-0.9916,P <0.001),the sensitivity was 83.3%,and the specificity was 80.0%.Conclusions:CHI3L1、PGLYRP-1、CEACAM8 participate in the occurrence and development of ESRD through inflammatory mechanism,which may be potential biomarkers of ESRD and provide new ideas for the diagnosis and treatment of ESRD.