Expression And Significance Of MicroRNAs In Plasma Exosomes Of Patients With Rheumatoid Arthritis

Background:Rheumatoid arthritis?RA?is a common autoimmune disease with symmetric polyarthritis as the main clinical manifestation,and the pathological changes are mainly synovitis.The etiology and pathogenesis of RA are complex,which is not clear at present.In recent years,some studies have confirmed that exosomes can participate in the pathogenesis of RA by delivering Micro RNA?miRNA?.Objective:By detecting the expression of miR-16-5p,miR-146a-5p,miR-155-5p and miR-223-3p in plasma exosomes of RA patients and healthy control group,to explore their significance in the diagnosis of RA diseases,and analyze the relationship between plasma exosomes miRNAs and clinical indicators of RA,so as to further understand their clinical value in RA diseases.Methods:A total of 56 RA inpatients from the Department of Rheumatology and Immunology of the Second Affiliated Hospital of Anhui Medical University from October 2017 to October 2019 were collected,including 28 in the active disease group and 28 in the stable disease group.At the same time,24 cases of healthy physical examination in the same period were collected as the control group.The exosomes of RA patients and healthy people were extracted by Qiagen exosomes extraction kit.Mi R-451a was used as internal control.The expression of miR-16-5p,miR-146a-5p,miR-155-5p and miR-223-3p in exosomes were detected by reverse transcription quantitative polymerase chain reaction technology.The data that conform to the normal distribution are expressed by`X±S,and others are represented by M(P₂₅,P₇₅).The t-test or rank sum test was used for comparison between the two groups of measurement data,and the chi-square test was used for comparison between the two groups of count data.ROC curves were used to analyze the diagnostic value of exosomal miRNAs for RA diseases.Results:?1?There was no significant difference in sex and age between RA group and healthy control group,RA disease activity group and stable group?P>0.05?.?2?Under the transmission electron microscope,the exosomes extracted from the plasma of healthy people and RA patients were round like vesicles with different sizes.The outer lipid bilayers were stained deeper,the inner low electron density materials were stained shallower,and the particle size was mostly distributed between 20-100nm.?3?There was no significant difference between RA group[660.381?581.212,807.308?]and healthy control group[675.897?559.328,752.316?],RA active group[634.963?561.095,756.107?]and RA stable group[676.374?589.167,898.333?)]?P>0.05?.?4?The expression of plasma exosomes miR-16-5p in the RA group[1.852?1.084,2.532?]was higher than that in the healthy control group[0.995?0.572,1.798?],the difference was statistically significant?P<0.05?.But the expression level of miR-16-5p in RA active group[1.427?0.950,2.314?]was comparable to that in RA stable group[1.970?1.248,2.857?],with no significant difference?P>0.05?.The ROC curve showed that AUC of miR-16-5p was 0.664?95%CI:0.536?0.792,P<0.05?.?5?The expression of plasma exosomes miR-146a-5p in the RA group[13.662?6.058,30.801?]was higher than that in the healthy control group[5.261?2.453,12.103?]?P <0.05?,and its expression in RA active group[27.542?13.905,87.017?]was higher than that in RA stable group[7.217?5.218,13.697?],and the difference was statistically significant?P<0.05?.The ROC curve showed that AUC of miR-146a-5p was 0.758?95%CI:0.649?0.867,P<0.05?.?6?The expression level of plasma exosomes miR-155-5p in the RA group[33.354?15.087,275.309?]was significantly higher than that in the healthy control group[11.684?1.551,17.221?],and the expression in RA active group[155.110?35.600,437.745?was also significantly higher than that in RA stable group[15.526?10.628,24.504?],and the differences were statistically significant?P<0.05?.The ROC curve showed that AUC of miR-155-5p was 0.806?95%CI:0.708?0.903,P<0.05?.?7?The expression level of plasma exosomes miR-223-3p in the RA group[106.611 ?46.813,223.841?]was significantly higher than that in the healthy control group [47.586?8.115,101.347?]?P<0.05?.Its relative expression level in RA active group[199.615?104.944,305.090?]was also significantly higher than RA stable group [53.993?21.524,107.707?],and the difference was statistically significant?P<0.05?.The ROC curve showed that AUC of miR-223-3p was 0.702?95%CI:0.581?0.823,P<0.05?.?8?There was no correlation between the relative expression levels of these four miRNAs in plasma exosomes of RA patients and age,sex,RF,anti-CCP antibody titer,and ESR?P>0.05?.The relative expression levels of miR-16-5p,miR-146a-5p and miR-223-5p had no correlation with CRP,while the relative expression levels of miR-155-5p had a certain correlation with CRP?P<0.05?.It was also found that the relative expression levels of miR-146a-5p,miR-155-5p,miR-223-5p were positively correlated with DAS28-CRP,DAS28-ESR,TJC,and SJC?P<0.05?.Conclusions:The abnormal expression of plasma exosomes miR-16-5p,miR-146a-5p,miR-155-5p and miR-223-3p in RA patients suggests that they are involved in the pathogenesis of RA and have the potential to become a new biomarker of RA.