Study On The Value Of Semaphorin3A In Early Diagnosis Of Chronic Kidney Disease

Objective:Semaphorin3A（Sema3A）,as a known secretory protein,is involved in angiogenesis,organ transplantation,cell migration,and cytokine release,which is associated with the occurrence of organs,immune responses and cancer.In the kidney,Sema3A is expressed in the glomeruli epithelial foot cell,distal convoluted tubule and collection tube,which is believed to play an important role in the regulation of renal development and function.Experimental studies have shown that the increased expression of Sema3A causes transient mass of proteinuria,apoptosis and endothelial injury.And Sema3A has been confirmed in a diabetes caused by glomerular disease and play a role,so far,it’s for CKD in clinical manifestations of chronic kidney disease,the role of study has not been clear.The purpose of this study was to clarify the relationship between Sema3A and CKD and explore its diagnostic value.Methods:In this study,96 patients with abnormal renal function or renal failure were selected from the general hospital of tianjin medical university and the second hospital of tianjin medical university from January 2014 to January 2016.The patients were aged between 22 and 89.In this study,each nephropathy was diagnosed according to renal biopsy and clinical manifestation and pathology report（immunofluorescence,optical microscopy and electron microscopy）.The patient’s disease was divided into thin basement membrane disease（TBM）,small lesion nephropathy（MCNS）,Ig-Anephropathy（IgA-N）,membranousnephropathy（MN）,diabet ic nephropathy（DN）,and focal segmental glomerulosclerosis（FSGS）.We collected a total of 76 biopsy specimens from five groups of patients outside the DN group.The serum and urine samples of all 96 patients were collected during their visit.The topic is divided into two parts.Part Ⅰ:The study is a retrospective analysis of multiple agencies,which assessed the serum and urine samples of 96 patients.Thirty healthy volunteers were also involved in the study.In addition,MCNS,iga-n and FSGS patients were followed up to detect the changes in the expression level of Sema3A in urine.Using enzyme-linked immunosorbent assay to determine the level of Sema3A in serum and urine,OD value was measured with microporous reader.The level of NAG in urine was measured by colorimetry.All the experimental data are expressed in standard deviation.The KW method is used for multiple comparisons.Spearman correlation coefficient is used to analyze the correlation between Sema3A and Albumin in urine.Wilcoxon signed rank test analyzed the changes in the level of Sema3A in patients with MCNS,IgA-N and MN in the onset.remission.P<0.05 was considered to be statistically significant.Part Ⅱ:This study used the kidney biopsy specimens from five groups of CKD withoutDN.The specimenswere paraffined,slicedandtreatedwith immunohistochemistry.The expression of Sema3A and NRP1 in different parts of the kidney was observed by optical microscope,and the semi-quantitative method was used to determine the positive cell staining,and the double blind method was used to score the cells.The correlation between Sema3A and NRP1 was analyzed by Spearman correlation coefficient.Results:（1）The urinary Sema3A,urine NAG,uALB,and serum Sema3A in the early CKD patients（all participating in the study）were increased compared with the control group.The urine Sema3A in each case group was compared with the control group,and the urine Sema3A level in each group was higher than that in the control group,among which the increase of MCNS group was the most obvious.Compared with the control group,the uALB group in each group was higher than the control group except TBM,and MN group was the most obvious.Compared with the control group,there was no statistical significance in TBM and MCNS group,and the NAG of IgA-N,MN,FSGS and DN was significantly increased,among which the increase of DN group was the most obvious.Compared with the control group,the serum Sema3A level of patients with MCNS and MN was higher than that in the control group,and the results were statistically significant.During the onset,the levels of urine Sema3A in patients with IgA-N,MCNS and MN were significantly higher than those in the remission period.The results were statistically significant,and there was no correlation between the other groups.In comparison,early CKD urinary Sema3A had no correlation with uALB.In the analysis of the correlation between urinary NAG and urine Sema3A,TBM group and MCNS group did not show correlation,the results no statistical significance,IgA-N,MN,FSGS group and DN groups were significantly positively correlated,the result was statistically significant,early chronic kidney disease（CKD）urine Sema3A has positive correlation with the urine NAG.The combined detection of early renal disease was higher than that of uALB and Sema3A,the area under the ROC curve was the largest.In comparison,AUC_combinedombined detection>AUCuALB,AUCcombineddetection>AUCSema3A,AUCSema3A>AUCuALB,andthe differences were statistically significant.（2）In each group,there were partial foot cells Sema3A staining and positive staining in the lumen of the distal tubules and collection tube cells in each group,while the proximal tubule was basically negative.The staining orientation of NRP1 and Sema3A was basically the same.Compared with the control group,there was statistically significant difference in the proportion of positive cells,the average staining score and the number of positive cases in the patients with nephropathy.In each group,NRP1 was compared with the control group,and the proportion of positive cells,the average color score,and the number of positive cases were statistically significant.The proportion of positive cells between Sema3A and NRP1 in each group was compared,the TBM group was 0.576,the MCNS group was 0.768,the iga-n group was 0.486,the MN group was 0.625,and the FSGS group was 0.379,which was positively correlated.Conclusions:The level of urinary Sema3A may reflect the extent of interstitial damage caused by glomerular injury.Meanwhile,urinary Sema3A may reflect the acute functional or morphological changes of the kidney in early CKD patients.Moreover,urine Sema3A may also reflect the degree of renal tubular damage in patients with iga-n,MN and FSGS.The diagnostic efficacy of Sema3A for chronic renal injury was higher than that of uALB,and the diagnostic efficacy of the combined detection was higher than that of the two.In the six early CKD studies,Sema3A and NRP1 were strongly positive in glomerulus,distal tubules and tubules.There was positive correlation between Sema3A and NRP1 in renal expression.