Relationship Between Interleukin-35 And Rheumatic Mitral Stenosis Disease

Objective:we aim to explore the changes of IL-35 in peripheral venous blood,the relative expression of mRNA ofp35 in peripheral venous blood mononuclear cells and the P35 protein concentration in diseased heart valves in patients with rheumatic mitral stenosis and whether the expression of P35 at different sites of the diseased valve is different Further analysis of the relationship exists between rheumatic mitral stenosis with different lesion levels and IL-35.We try to find the role of IL-35 in the progression of rheumatic mitral stenosis,to eplore whether IL-35 can become a new direction for the treatment of rheumatic valvular heart disease.Methods:In this study,we selected mitral stenosis-based rheumatic valvular heart disease patients and classified them into moderate and severe mitral stenosis groups based on the degree of mitral stenosis,including 20 cases of moderate mitral stenosis patients,20 patients with severe mitral stenosis,20 healthy controls,and 15 patients with mitral valve prolapse.Among them,all patients underwent mitral valve replacerment.Prospective collection of peripheral venous blood and four sites of mitral valve tissue,labeled as P1,p2,p3,p4,from all enrolled subjects was performed.Enzyme-linked immunosorbent assays were used to determine the amount of IL-35 in peripheral vein plasma and Western blotting was used to determine the relative expression of P35 subunits in the valve tissue.Quantitative fluorescent polymerase chain reaction was used to detect the relative expression of mRNA of P35 in peripheral blood mononuclear cells.Results:In the plasma of peripheral venous blood,the levels of IL-35 in the normal control group,the moderate and severe mitral stenosis group were 60.46±9.83,100.1±16.34 and 110.4±14.64(pg/ml),respectively.Importantly,significant increase of IL-35 cytokine was observed in moderate mitral stenosis patients(p<0.05)and severe mitral stenosis patients(p<0.01)as compared healthy subjects(60.46±9.830 pg/ml),and there was no statistical difference between the two groups of moderate and severe mitral stenosis.In addition,we also comepared healthy subjects and RMS patients,and the result showed that IL-35 cytokine in RMS patients was(105.2±10.86 pg/ml,n=40,P= 0.0103<0.05)higher when compared healthy subjects(60.46 ± 9.830pg/ml n=20);In this study we took mitral valves from prolapse patients as contral because healthy mitral valves are unattainable.Among all the RMS patients(n=15),high amounts of P35 were detected in the mitral valve tissue(P1 locus)specimens compared with the contral patients(P1 locus)(p=0.0109<0.05,n=15).Further analysis p35 in both moderate mitral stenosis patients(P=0.0476<0.05,n=7)and severe mitral stenosis patients(p=0.0483<0.05,n=8,),were higher than in contral patients,but there is no significance between mitral stenosis patients and severe mitral stenosis patients(p=0.6587);In the valve tissue of rheumatic mitral stenosis,the expression of P35 in the valve tissues of different sites(P1,P2,P3,P4)was not statistically different(P>0.05,n=4).;Compared with the normal control group(n=20),the relative expression ofp35 mRN A in peripheral blood mononuclear cells in the patients with moderate mitral stenosis(n=20)and severe mitral stenosis(n=20)were increased by about 2-fold and 2.4-fold respectively,with both p-values<0.001,and There was no statistical difference between the moderate and severe groups,(p=0.1780).The p35 mRNA expression was increased in patients with mitral stenosis group(n=40)conpared with the normal control group(n=20)(p<0.01);According to Spearman’s correlation analysis,IL-35 concentration in plasma,and the expression of p35 mRNA in blood mononuclear cells was positively correlated with the degree of mitral stenosis(r=0.453,p<0.01,n=60 and r=0.549,p<0.01,n=60).Conclusion:In patients with rheumatic mitral stenosis,the plasma levels of IL-35 and perpheral blood mononuclear cells in patients with mitral stenosis were increased,and the increase was positively correlated with the degree of mitral stenosis.The expression of p35 protein in valve tissue of patients with rheumatic mitral stenosis was higher than that of patients with mitral valve prolapse.