| Objective Rheumatoid arthritis(RA)is a chronic systemic autoimmune disease.The main pathological changes include synovitis,bone and cartilage destruction,pannus formation,resulting to progressive joint injury.Fibroblast-like synovitis(FLSs)are the most important functional cells which are characterized with abnormally proliferated and activated.In recent years,more and more studies have found that exosome-derived circ RNAs are stably expressed in body fluids such as plasma and urine,and have a variety of biological functions.They are important regulatory factors of epigenetic gene expression in inflammation and autoimmune regulation.They are closely related to the occurrence and development of rheumatoid arthritis and can be used as biomarkers.The purpose of this study was to investigate whether the expression of plasma exosomederived circ RNAs in RA patients was different from that in healthy controls,and its relationship with clinical indicators and medication in RA patients.In terms of basic research,we preliminatively explored the effect of circ RNA significantly expressed in RA plasma exosomes on proliferation and migration of RA FLSs.Methods A total of 60 inpatients and outpatients was diagnosed with RA in the rheumatology department of Affiliated Hospital of Nantong University from September2020 to December 2022 were collected,and 34 healthy controls underwent health examinations in this hospital during this period.The plasma sample,clinical test results as blood routine,biochemistry,procalcitonin(PCT),erythrocyte sedimentation(ESR),Creactive protein(CRP),rheumatoid factor(RF),anti-cyclic citrullinated peptide antibody(anti-CCP antibody)in RA patientsbasic were collected.The drug use,gender,age and other basic information of RA patients during treatment were recorded.Plasma exosomes were extracted by gradient ultra-high-speed centrifugation.The extracted plasma exosomes were identified by using Zataview particle size tracking analyzer to analyze the particle size distribution of exosomes and using Western-blot to verify the expression of exosome characteristic protein.Gene sequencing and biogenetic analysis showed that has＿circ＿0003914 was significantly expressed in RA plasma exosomes.The sample size was further expanded and q RT-PCR was used to verify its expression in plasma.By transfection overexpressed plasmid/silencing si RNA into RA-FLSs,the expression of hsa＿circ＿0003914 was overexpressed/inhibited.Subsequently,CCK-8 and scratch tests were performed to investigate the effects of plasma exosomes-derived hsa＿circ＿0003914on proliferation and migration of RA-FLSs.SPSS was used to analyze the correlation between the expression level of has＿circ＿0003914 and clinical data of RA patients.Independent sample t test,Mann-Whitney U test and Pearson/Spearman test were used for statistical analysis.Results Gene sequencing and biogenic analysis showed that hsa＿circ＿0003914 was specifically highly expressed in plasma exosomes of RA patients.The expression of hsa＿circ＿0003914 in RA plasma was significantly higher than that in healthy control group(P=0.033).The expression level of hsa＿circ＿0003914 was correlated with age,VAS,Hb,Plt,PCT and complement C3C4 of RA patients(P<0.05),and it was no correlation with disease course,DAS28,ESR,CRP,RF and related-antibodies.And It was negatively associated with glucocorticoid and tripterygium wilfordii use(P<0.05).The results of cell experiments showed that the proliferation and migration of RA-FLSs could be regulated by regulating the expression level of hsa＿circ＿0003914.Conclusions Studies have shown that hsa＿circ＿0003914 has different expression in plasma exosomes and plasma between RA patients and healthy controls,which can be used as a potential marker of RA and has certain clinical application value.Basic studies have found that regulating the expression level of hsa＿circ＿0003914 can regulate the proliferation and migration ability of RA-FLSs,but the specific mechanism needs to be further studied. |
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