Effects Of CD14 And CD16 Monocytes On Calcified Aortic Valve Disease

The first part used bioinformatics to determine the degree of infiltration of immune cells in calcified aortic valvesObjective To explore the effect of immune cells on calcification during the development of CAVD.Method Using GEO database,download immune cells infiltrating genes associated with aortic valve calcification data,isolated from patients get chip annotation file the corresponding probe matrix,using systems biology tools CIBERSORT deconvolution complex algorithm for large gene expression data,solution of convolution by microarray gene expression data analysis of immune cells.Get the calcification of aortic valve group and normal group 22 kinds of immune cells infiltration in the corresponding matrix,screening in calcified valve and normal tissue immune cells compared three groups of related data sets GSE51472,GSE12644 and GSE83453,by using Wilcoxon filtered nonparametric test of three groups of samples,obtained differentially expressed immune cells to(p< 0.05),which expression significance for screening of samples.Results Data set GSE51472 showed that plasma cells,macrophage M0,macrophage M2,regulatory T cells(Treg),T cells and natural killer activated cells all had different expressions,but the content of mononuclear cells in calcified valve tissue was lower than that in normal valve tissue.Data set GSE12644 showed that mononuclear cells,macrophage M0 and natural killer activated cells had different expressions.GSE83453 data set shows that mononuclear cells and dendritic resting cells have different expressions.At the same time,x CELL algorithm was used to identify the differential expression of immune cells,so as to show the difference(p<0.05).The results showed that mononuclear cells were expressed in three sets of data,among which two groups showed differences.The content of mononuclear cells in calcified valve tissues in the three groups was lower than that in normal valve tissues,and all showed a decreasing trend,indicating that mononuclear cells may be involved in the progression of aortic valve calcification.Conclusion It is found that immune cells play a role in the progression of valvular calcification,and the content of mononuclear cells varies in the changes of valve tissues.Mononuclear cells may provide some research basis for the progression of valvular calcification.In the second part,the results of the differential expression of mononuclear cells in the data set were verifiedObjective To collect human valve tissue and verify whether the infiltration of mononuclear cells in calcified valve tissue is consistent with the analysis results of GEO data set.Real-time PCR and immunohistochemistry were used to detect the expression of mononuclear cells in the valve tissue,and the influence of mononuclear cells on the progression of valve calcification was analyzed.Methods 7 cases of calcified valve group and 7 cases of normal tissue,the expression of the data set has the difference of mononuclear cells for validation,validation of monocyte specific subtype of CD14 monocyte and CD16 expression of monocytes in calcified valve and normal level,to detect the contents of mononuclear cells in calcified valvular tissue,using the Real-time PCR detection CD14 monocyte and CD16 mononuclear cells in the expression of calcified valve group and normal group,immunohistochemical detection of mononuclear cell staining,observed in infiltration expression of calcified valve group and normal group.It provides a theoretical basis for studying the effect of mononuclear cells on CAVD.Meanwhile,the effects of different clinical indicators on the progression of aortic valve calcification were analyzed based on the clinical information of patients.Results Quantitative analysis of valve tissue shows that the expression of CD14 monocytes in calcified valve tissue is lower than that in normal valve tissue,which is consistent with the result that the infiltration expression of monocytes in calcified group is lower than that in normal group.CD14 monocytes are mainly involved in the progress of valve calcification,and the lower the content,the more serious the degree of valve calcification.Conclusion CD14 monocytes may be related to the calcification progress of CAVD.The infiltration density of monocytes in normal valve tissue is significantly higher than that in calcified valve tissue.However,the expression of CD16 monocytes in calcified valves is not significantly different from that in normal valve tissues.