Association Study Of RNA-binding Protein For N6-Methyladenosine With Systemic Lupus Erythematosus

ObjectivePeripheral blood mononuclear lymphocytes(PBMC)were collected from SLE and healthy controls.The expression level was measured at the m RNA level.We analyzed the differences in expression levels of m6 A binding protein in SLE patients and healthy controls,and discussed the relationship between m6 A binding protein expression levels and the clinical characteristics of SLE,the expression levels of SLE related cell secretory cytokine.Otherwise,we discussed the diagnostic value of m6 A binding protein for SLE.MethodsIn this study,a case-control study design was adopted to collect peripheral venous blood from SLE patients and healthy control population,and monocytes and plasma samples from peripheral blood samples were extracted within 4 hours for subassembly.The total RNA reverse transcription was extracted from monocytes as c DNA library,and the m RNA expression levels of the proteins involved in M6 A modification recognition were detected by quantitative PCR assay.ELISA assay was made to detect the expression levels of specific cell secretory cytokine in plasma samples in SLE patients and healthy control.(1)The difference in expression level of m6 A binding protein between SLE patients and healthy controls was analyzed;(2)To investigate the correlation between the expression level of m6 A binding protein and its clinical situations in patients with SLE;(3)To study the correlation between the abnormally expressed m6 A binding protein and the expression levels of related plasma secretory factors,and to provide certain theoretical basis for the pathogenesis and related treatment of SLE patients.Results(1)PCR assay showed that m RNA expression levels of the m6 A binding proteins YTHDF1 and HNRNPA2B1 were different between SLE patients and healthy controls,(Z=-2.221,P=0.026;Z=-2.427,P<0.001).There was no significant difference in other m6 A binding proteins between the patients and the healthy control group.YTHDF1 was correlated with the expression levels of YTHDF2 and YTHDF3.There was correlation between YTHDF2 and YTHDC1 and YTHDC2.YTHDF3 was correlated with YTHDC2 and HNRNPA2B1.There was also a correlation between YTHDC2 and HNRNPA2B1.(2)The expression level of YTHDF1 was correlated with arthritis,low complement,prothrombin time,and YTHDF2 was correlated with immune protein Ig G,Ig M,rheumatic index erythrocyte sedimentation rate(ESR),antistreptococcal hemolysin(ASO),and absolute CD8.YTHDC1 was positively correlated with immune protein Ig M and rheumatic index ESR.YTHDF3 was positively correlated with INR and FIB.The binding protein was correlated with the level analysis of some clinical biochemical indexes in patients with SLE.The diagnostic value of the binding protein HNRNPA2B1 as a potential marker for the diagnosis of SLE disease was 0.790(95%CI: 0.704-0.876).(3)The differences in the expression levels of cytokines IL-2,IFN-a and IP-10 between the patients and the healthy control group were analyzed,and only the differences of IP-10 between the patients and the control group were statistically significant.according to clinical manifestations related to clinical disease activity,there was no significant difference in the expression levels of cytokines between the two groups.The expression level of plasma factor IFN-a in anti RNP antibody(r RNP)positive and negative groups was significantly different.Cytokine IL-2 was associated with positive anti-nucleosome antibodies(Anu A)and immunoprotein Ig A levels.Cytokine IP-10 was positively correlated with rheumatic index hypersensitive C-reactive protein(hs CRP).(4)The expression level of YTHDF1 was positively correlated with the expression level of IFN-a(rs=0.337,P=0.048),and negatively correlated with the expression level of IP-10(rs=-0.307,P=0.048).The m6 A binding protein YTHDF3 was positively correlated with the expression level of cytokine IL-2(rs=0.443,P=0.007).The expression level of m6 A binding protein YTHDF3 was negatively correlated with cytokine IP-10(rs=-0.442,P=0.003).Conclusions(1)The m RNA expression levels of m6 A binding protein YTHDF1 in peripheral blood monocytes and lymphocytes of SLE patients were lower than those in healthy controls and HNRNPA2B1 were higher,the difference was statistically significant.(2)The m6 A binding protein was correlated with some clinical situations of SLE.The m6 A binding protein HNRNPA2B1 has high diagnostic value for SLE disease and can be used as a potential biomarker for SLE disease.(3)The correlation between some m6 A binding protein and the expression levels of IL-2,IFN-a and IP-10 involved in immune regulation of autoimmune diseases was statistically different,and the mechanism of their involvement in immune regulation needs to be further explored.