Expression And Correlation Of Interleukin(IL)-35 In Rheumatic Mitral Stenosis

Objective:From the morphological point of view,in the patients with rheumatic valvular heart diseasewhichwas diagnosed as RHVD for mitral stenosis as the main lesion,the expression of IL-35 in the mitral valve with different degrees of stenosis,the expression of IL-35 in different parts of the same valve tissue;To explore the changes ofthe proportion between the Treg cells and Breg cellsof the peripheral blood venous mononuclear lymphocytes(PBMC)which can expresse IL-35 in patients with rheumatic valvular heart disease,and Further analysis that there is a correlation between mitral stenosis and IL-35.To explore the role of anti-inflammatory factor IL-35 in the progression of rheumatic mitral stenosis,and to provide reference for the early treatment of IL-35 as a prophylactic valvular heart disease.Methods:In the morphological experiment,we selected rheumatic mitral stenosis as the main lesion in patients with rheumatic valvular heart disease as the experimental group,according to the degree of mitral stenosis into two groups of moderate and severe mitral stenosis,respectively 19 patients and 24 patients,and 6 patients with congenital mitral valve prolapse were included in the control group.All patients underwent mitral valve replacement or mitral valvuloplasty.The diseased valves replaced by the patients were collected,and the valve tissues of four different sites were taken according to different parts,labeled as P1,P2,P3 and P4,respectively.The semi-quantitative and immunofluorescence co-localization methods were used to analyze the immunohistochemistry.The expression of IL-35 in the valve tissue of the experimental group and the control group.In addition,peripheral venous blood was collected from healthy volunteers(n=6)and patients(n=8),and the trend of the ratio of Treg and Breg in the peripheral venous blood of the experimental group and the normal group was compared by flow cytometry.Results:In immunohistochemical staining of patients with rheumatic heart disease,cells expressing two subunits of IL-35,EBI3 and p35,showed varying degrees of sepia coloration in the cytoplasm.In the rheumatic mitral mid-range(n=5),severe stenosis(n=4)and prolapse control group(n=4),there was no significant difference in the expression levels of EBI3 and p35 between different parts of the valve(P1,P2,P3,P4).P>0.05,n=5).Furthermore,the same site(PI)of all valve tissues was selected and found to be in patients with severe rheumatic mitral stenosis(n=24),patients with moderate mitral stenosis(n=19),and patients with congenital prolapse(n=6).In the valve tissue,the positive rates of the EBI3 subunit were 10.24±0.5755(%),7.076 ± 0.7798(%),and 5.784 ± 0.6455(%),respectively,in the severe stenosis group and the moderate stenosis group and the prolapse group,respectively.The comparison was significantly increased,p values were pi=0.0018<0.05,p2<0.0001,and there was a statistical difference.However,compared with the prolapsed group,p=0.2433>0.05,no statistical difference;In severe and moderate stenosis,Spearman's grade correlation analysis was performed in three groups of degree stenosis and prolapse group,p<0.001,and correlated.The positive rates of p35 subunits were 10.90:±0.8605(%),6.630±0.4736(%),and 6.129 ± 0.5345(%),respectively.Similarly,the severe stenosis group was significantly higher than the moderate stenosis group and the prolapse group.The p values were pi=0.0002<0.05,p2=0.0005<0.05,and there was a statistical difference.In the moderate and prolapsed groups,p=0.4942>0.055 no statistical difference;in severe stenosis,moderate stenosis and off Spearman's grade correlation analysis was performed in the three groups of the vertical group,p<0.001,with a correlation.Flow cytometry revealed that the proportion of Treg cells and Breg cells in peripheral blood PBMC of RHD patients was higher than that of normal group,pi=0.0043<0.05(n=8),p2=0.0007<0.05(n=6),all have statistical differences;and the increase in Breg cells compared with Treg cells was found to be significant(p<0.0001).Conclusion:In patients with rheumatic mitral stenosis as the main lesion,there was no difference in expression between four different sites(P1,P2,P3,P4)in the same valve tissue;however,in different mitral stenosis groups,The expression levels of EBI3 and p35 protein in the two subunits of IL-35 in the mitral stenosis were significantly higher than those in the moderately stenotic and prolapsed valve tissues,and their expression levels were positively correlated with the degree of lesions.In peripheral blood PBMC of patients with rheumatic mitral stenosis,the proportion of Treg and Breg cells was higher than that of the normal control group,and the proportion of Breg cells increased significantly compared with the proportion of Treg cells.