Study On Differential Expression Of MiRNA And MRNA Related To Tibetan SLE In Diqing Area Of Yunnan Province

Objective:This study explored differentially expressed miRNAs and mRNAs in peripheral blood mononuclear cells of Tibetan patients with systemic lupus erythematosus in Diqing area of Yunnan Province.Based on bio informatics analysis methods,a miRNA-mRNA regulatory network was constructed,miRNAs and target genes of interest were screened,and their mechanisms and disease-related biomarkers involved in the development of lupus were explored in order to guide the diagnosis and treatment of SLE in clinical practice.This study is expected to fill a gap in SLE-related research of the Tibetan population in Yunnan Province.At the same time,it may provide a theoretical basis for further research on the pathogenesis and rational prevention and treatment of environmental and genetic factors in SLE.Method:This study collected 20 ml of fasting peripheral venous blood from 10 Tibetan SLE patients(SLE group)and 10 Tibetan healthy controls(NC group).PBMC was isolated and RNA was extracted.In the SLE group and the NC group,5 patients were randomly selected from each group for high-throughput sequencing to obtain miRNA and mRNA expression profiles differentially expressed in SLE patients compared with healthy controls.After further verifying the authenticity of the sequencing results by fluorescent quantitative PCR,GO enrichment analysis and KEGG pathway enrichment analysis of differential genes,construction of PPI network,and screening of core genes were performed.Subsequently,the target genes of the miRNAs that have been screened and the differential mRNAs are intersected to obtain the intersected genes.And it was analyzed by GO enrichment analysis and KEGG pathway enrichment analysis,and finally the miRNA-mRNA regulatory network was constructed.Screening differential miRNAs and differential genes that play an important role from the miRNA-mRNA regulatory network.Results:1.Second-generation high-throughput seq uencing screened a total of 45 significantly differentially expressed miRNAs,including 13 up-regulated and 32 down-regulated;469 significantly differentially expressed mRNAs,including 356 up-regulated and 113 down-regulated.2.RT-qPCR detected the expression of miRNA in PBMC of 10 Tibetan SLE patients and 10 Tibetan healthy controls.The results showed that:compared with the healthy control group,miRNA-3942-3p and miRNA-3614-3p in SLE patient group the expression level of miRNA in PBMC was up-regulated,and the difference was statistically significant;miRNA-4512 and miRNA-4434 were down-regulated in the PBMC of SLE patient group,and the difference was statistically significant The results of the four miRNA RT-qPCR are basically consistent with the high-throughput sequencing results,indicating that the sequencing results are credible.3.The GO analysis of differential genes is enriched in biological processes that may be related to lupus,such as immune response,inflammatory response,cell chemotaxis,lymphocyte activation,Toll-like receptor signaling pathway regulation,chemokine secretion and regulation.KEGG pathway analysis obtained lupus-related chemokine signaling pathway,IL-17 signaling pathway and other pathways.15 core genes were selected from the constructed PPI network.4.In the miRNA-mRNA regulatory network,the largest contribution is miR-4512,and multiple differential miRN As and mRNAs that play an important role are screened out,including 2 miRNAs(miR-125b-5p,miR-31)and 4 mRNA(LPAR1,CXCL5,CXCL13,and CXCL16)are currently reported to be related to lupus.Conclusion:1.There are multiple microRNAs and mRNAs expression differences in PBMC of Tibetan SLE patients in Diqing area of Yunnan Province compared with normal Tibetans.2.The onset of SLE is a complex biological process,and there are many types of genes and pathways involved in its occurrence and development.3.In the miRNA-mRNA regulatory network,the most important contribution is miR-4512,and multiple differential miRNAs and mRNAs selected from it(miR-125b-5p,miR-31,LPAR1,CXCL5,CXCL13 and CXCL16 Etc.).They may participate in the occurrence and development of SLE,and provide new ideas for further in-depth exploration of the pathogenesis of Tibetan SLE in high-altitude areas,and for finding potential biomarkers and targeted treatments for diseases.