| Objective: In this study,high throughput sequencing technology was used to screen the lncRNA and miRNA biomarkers in the peripheral blood of patients with rheumatoid arthritis,and GO function enrichment analysis and KEGG pathway enrichment analysis were carried out on the lncRNA and miRNA differentially expressed genes in the patients with RA treated by the three-dimensional comprehensive therapy of Zhuang Medicine.The prediction of biological processes and signaling pathways involved in the source genes of differentially expressed lncRNA and miRNA,it will provide a preliminary scientific basis for exploring the mechanism of three-dimensional comprehensive therapy of Zhuang medicine in the treatment of RA,and highlight the characteristics and advantages of Zhuang medicine in preventing and treating RA,and provide feasible ideas and methods for the biological basis research of Zhuang medicine in the prevention and treatment of major diseases.Methods: 10 patients with RA who were treated in the Department of Rheumatology of the First Affiliated Hospital of Guangxi University of Chinese Medicine from January 2020 to June 2020,and it randomly divided into control group and observation group,5 cases in each group.In addition,5healthy people who underwent physical examination in the First Affiliated Hospital of Guangxi University of Chinese Medicine during the same period were recruited as the normal group.The observation group and the control group were treated as follows:(1)methotrexate tablet once a week,10 mg each time;(2)Leflunomide tablets once a day,10 mg each time;they are as the treatment basis.The control group was given Longzuan Tongbi granules of Zhuang medicine(15g/ bag)orally,2 times a day,1 bag each time;and 4weeks as a course of treatment,a total of 2 courses of treatmen.On the basis of western medicine treatment,the observation group was treated with Zhuangyi three-dimensional comprehensive therapy(The combination therapy consists of internal administration of Longzuan Tongbi granules,and external use of zhuang medicine heat sensitive acupuncture point therapy,and cooperate to take strong medicine hair prosperous food therapy prescription),they were treated 4 weeks as a course of treatment,a total of 2 courses of treatment;The normal group received no treatment.During the treatment,5m L peripheral venous blood was extracted from patients with rheumatoid arthritis before and after treatment,and was extracted from healthy subjects on the second day of recruitment.Lnc RNA and miRNA were detected by high-throughput sequencing technology.The sequencing data were processed by R 3.6.1,Fast QC 0.11.7,Stringtie 1.3.3 and other software.Go enrichment analysis and KEGG pathway enrichment analysis were performed on the selected lncRNA and miRNA source genes with differential expression by David database.Results: 1.Screening results of lncRNA markers:(1)Compared with the normal group and the rheumatoid arthritis group,10,199 lncRNAs were differentially expressed and 1779 lncRNAs were significantly differentially expressed,including 50 lncRNAs up-regulated and 1729 lncRNAs down-regulated.(2)Compared with the observation group before and after treatment,9112 lncRNAs were differentially expressed and 249 lncRNAs were significantly differentially expressed,including 69 lncRNAs up-regulated and 180 lncRNAs down-regulated.(3)Compared with the control group before and after treatment,7692 lncRNAs were differentially expressed,and 122 lncRNAs were significantly differentially expressed,including 76 lncRNAs up-regulated and 46 lncRNAs down-regulated.(4)After treatment,8810 differentially expressed lncRNAs were found in the observation group and the control group,among which 659 were significantly differentially expressed,627 were up-regulated and 32 were down-regulated.2.Effects of three-dimensional comprehensive therapy in Zhuang Medicine on the differential expression of lncRNAs:(1)In the co-expression network of lncRNA-m RNA,there were 118 nodes and 365 edges.AC073548.1 was the m RNA with the most co-expression,becoming the core regulation module of the co-expression network and the most important lncRNA in the network.(2)A total of 1110 items were significantly enriched in GO items of target genes with differentially expressed lncRNAs,including878 items for Biological process,146 items for Cellular component and 86 items for Molecular function.(3)Hematopoietic cell lineage,NF Kappa B signaling pathway,and Osteoclast differentiation are the pathways that significantly enriched the differentially expressed lncRNA target genes in KEGG analysis.3.Results of screening of miRNA markers:(1)Compared with the normal group and the rheumatoid arthritis group,a total of 47 miRNAs were significantly differentially expressed,including 25 miRNAs target genes were up-regulated and 22 miRNAs target genes were down-regulated.(2)A total of56 miRNAs were found in the observation group before and after treatment,among which 27 were up-regulated and 29 were down-regulated.(3)Compared with the control group before and after treatment,there were 14 significant differences in miRNAs expression,among which 1 was up-regulated and 13 were down-regulated.(4)Compared with the control group,19 miRNAs were significantly changed in the observation group after treatment,including 8up-regulated and 11 down-regulated.4.The effect of three-dimensional comprehensive therapy in Zhuangming Medicine on the differential expression of miRNA:(1)A total of1651 items were obtained by GO enrichment analysis of up-regulated target miRNA genes,including 1289 items for Biological process,174 items for Cellular component and 188 items for Molecular function.The pathway of significant enrichment in the KEGG analysis involved melanoma,actin cytoskeleton regulation,pathogenic E.coli infection,MAPK signal transduction pathway,and herpes simplex virus infection,etc.(2)A total of2293 entries were obtained by GO enrichment analysis of down-regulated miRNA target genes,including 1770 entries for Biological process,335 entries for Cellular component,and 188 entries for Molecular function.Glioma,PI3K/Akt signaling pathway,endocrine antagonism,and small molecule ri RNA in tumors were the pathways that significantly enriched the differentially expressed target miRNA genes in KEGG analysis.Conclusion:(1)There are a large number of differentially expressed lncRNAs and miRNAs in the peripheral blood of patients with rheumatoid arthritis.These differentially expressed lncRNAs and miRNAs are closely related to the occurrence of RA and may become biomarkers for the diagnosis of RA.(2)Stereoscopic comprehensive therapy in Zhuang medicine may affect the inflammatory response and immune response of the body through the relevant differentially expressed lncRNAs to participate in the prevention and treatment of RA.(3)The three-dimensional comprehensive therapy of the Zhuang medicine may up-regulate the expression level of hsa-mi R-133 p,hsa-mi R-206,hsa-mi R-1-3p,and down-regulate the expression level of hsa-mi R-9-5p,hsa-mi R-124-3p,hsa-mi R-129-5p,and hsa-mi R-129-5p,and It may be associated with MAPK signaling pathway,Hippo signaling pathway,PI3K/Akt signaling pathway and other signaling pathways to play a therapeutic role. |
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