Differential Expression Of MicroRNA In Patients With Rheumatoid Arthritis Of Cold Syndrome And Heat Syndrome

Objective: This project aims to explore the nature of cold syndrome and heat syndrome of Rheumatoid Arthritis at the level of micro RNA by studying the difference of RA Gene expression profiling with cold syndrome and heat syndrome by using the second generation sequencing technology,and further excavate the susceptible genes that have been found,to provide a strong basis for improving the effect of TCM syndrome differentiation and treatment of RA.Methods: By Illumina high-throughput sequencing platform,the expression levels of miRNA in peripheral blood mononuclear cells(PBMC)were detected in 10 patients with cold syndrome(cold-dampness-arthralgia syndrome),5 patients with heat syndrome(dampness-heat-arthralgia syndrome)and 5 healthy volunteers,the miRNA differentially expressed genes between RA and healthy volunteers with cold and heat syndrome were screened by bioinformatics analysis,and 40 RA patients with cold syndrome and 33 RA patients with heat syndrome were selected,seven genes representing the difference between cold and hot syndromes of RA were selected and polymerase chain reaction by quantitative real-time polymerase chain reaction(q RT-PCR).Results: The laboratory indexes showed that the PLT,ESR,CRP,IgA,IgG of the patients with RA heat syndrome were higher than those of the patients with cold syndrome.The results of sequencing showed that 49 genes were up-regulated and 60 genes were downregulated in RA cold syndrome patients,while 15 genes were up-regulated and 32 genes were down-regulated in RA heat syndrome patients.Gene function enrichment analysis showed that the differential miRNA genes of RA cold syndrome were mainly involved in the regulation of cellular processes,the regulation of primary metabolic processes and development,and played their roles through MAPK and insulin signaling pathways The differential miRNA genes of RA heat syndrome are mainly involved in the regulation of cellular processes,developmental processes,anatomical structure development and other biological processes,and play their functions mainly through autophagy and TGF-BETA signaling pathway.q RT-PCR results showed that hsa-miR-1827 was down-regulated in cold syndrome(P = 0.004),hsa-miR-651-5p was up-regulated in cold syndrome(P=0.031),and hsa-miR-133 b,hsa-miR-133a-3p was up-regulated in heat syndrome(P = 0.005,P=0.010),hsa-miR-96-5p was down-regulated in both cold and heat syndromes(P= 0.003).Conclusion: The levels of PLT,ESR,CRP,IgA and IgG in the RA heat syndrome group were higher than those in the cold syndrome group.There were significant differences in Mirna between cold and heat syndrome and healthy people.hsa-miR-1827 and hsa-miR-651-5p could be used as biomarkers of cold syndrome of RA,and hsa-miR-133b、hsa-miR-133a-3p could be used as biomarkers of heat syndrome of RA.