| Objective: Membranous nephropathy(MN)is one of the most common pathological types of adult nephrotic syndrome.Membrane nephropathy may be divided into idiopathic membranous nephropathy(IMN)and secondary membranous nephropathy(SMN)according to whether it has a clear cause.At present,idiopathic membranous nephropathy is associated with podocyte autoimmune diseases,under certain conditions leads to podocyte injury and apoptosis,resulting in a large number of proteinuria.Type 1 thrombospondin type-1 domain-containing 7A,THSD7 A was originally thought to be expressed in placental vascular endothelial cells.Tom Tomas et al found that62 patients with idiopathic membranous nephropathy in 15 patients with serum in the presence of a new different from the human phospholipase A2receptor(PLA2R)circulating antibody,named Type 1 thrombospondin type-1domain-containing 7A,and is the second antigen involved in the onset of adult IMN.The aim of this study was to investigate the expression of THSD7 A in renal tissue of IMN patients,to detect the expression of THSD7 A in blood samples of IMN patients,and to explore the mechanism of THSD7 A in the pathogenesis of IMN.Methods:1 Object of studyA total of 60 patients with glomerular disease were enrolled in our hospital from October 2014 to June 2015.Among them,33 males and 27 females had an average age of(42.6 ± 15.8)years.(2)There were 10 cases of SMN group,5 cases of male and 5 cases of female,the average age(36.1 ±14.0 years),and the average age was(41.6 ± 11.2)),7 cases of hepatitis B virus-associated membranous nephropathy(HBV-MN),4 males and 3 females,mean age(43.6 ± 11.1)years;3 cases of V-type lupus nephritis(LN-V),male1(MCD)was selected in 6 cases,6 males and 6 females were randomly divided into two groups: the control group(n = 6)and the control group(n =6).The mean age(38.9 ± 18.2)Female 4 cases,the average age(30.2 ± 16.5)years old,as the research group of the study group,all selected candidates have signed informed consent,and by the hospital ethics committee to discuss the adoption.2 Collect general information Including patient sex,age,height,weight,blood pressure,current history,past history and so on.3 Collect specimens Collected celiac day fasting venous blood 5ml,3500 r / min centrifugal10 min(centrifugal radius of 13.5cm),take the supernatant stored at-80 ?for inspection.B-guided renal biopsy,renal tissue by formaldehyde-acetic acid-ethanol fixative(FAA)fixed 24 h,gradient ethanol dehydration,xylene transparent,impregnated with wax.4 Detection of THSD7 A in renal tissueAll tissues were sacrificed by immunohistochemistry for THSD7 A staining.The kit was purchased from Beijing Boao Sen Biotechnology Co.Ltd.The immunohistochemical staining of the immune tissue is performed according to the experimental procedure provided by the kit.The results showed that dark brown granules were positive along the glomerular basement membrane.Negative control: PBS instead of primary antibody as a negative control,only the addition of PBS negative and add a positive anti-positive,was judged to be positive.5 Detection of serum THSD7 A antibody(THSD7A-AB)Using enzyme-linked immunosorbent assay,kit purchased from Shanghai Xitang Biotechnology Co.,Ltd.,according to the instructions for the steps.6 Detection of serum THSD7 A antibody(THSD7A-AB)Serum albumin,serum creatinine,urea nitrogen,uric acid,total chole-sterol,autoantibody,hepatitis B and 24 h urinary protein were measured by automatic biochemical analyzer in our hospital.7 Statistical methods The data were analyzed by SPSS18.0 software.The experimental data were expressed as mean ± standard deviation(sx ±),count data rate or composition ratio,and the mean distribution of multiple data was analyzed by variance analysis,correlation test Linear correlation analysis,with P <0.05 for the difference was statistically significant.Results:1 General clinical data and serum THSD7A-AB expression in MN patientsThere was no significant difference in age,sex composition,systolic blood pressure,body mass index and urinary protein between the two groups(P> 0.05).The results showed that serum anti-THSD7 A antibody was positive in 14(35%)IMN patients and the titer was high(mean 108.38 ng / ml).Serum anti-THSD7 A antibody was positive in 2(20%)SMN patients with titer of 19.28 ng The positive rate of THSD7 A antibody in blood was significantly higher than that in the other two groups,the difference was statistically significant(P <0.05).2 Expression of THSD7 A in renal tissueTHSD7A positive expression in the glomerular dark brown along the capillary loop granular deposition,the positive expression of each group the same site.The positive rate of THSD7 A in IMN group was significantly higher than that in SMN group(P <0.05),but there was no expression in MCD group(P <0.05)Figure 1,and HBV-MN and LN-V group between the difference was not statistically significant(P> 0.05).3 Correlation between THSD7 A and serum antibody levels in renal tissue of patients with IMN and clinical(P <0.05).There was no significant difference in age,total cholesterol and Scr between the two groups(P> 0.05),and there was no significant difference between the two groups(P> 0.05),and the serum levels of THSD7A-AB Table 2.(R =-0.286,P <0.001;r = 0.564,P<0.001),and there was no correlation with total cholesterol and Scr(r =0.564),and the positive rate of THSD7 A in renal tissue of IMN was correlated with albumin and urinary protein(R = 0.359,P = 0.035;r = 0.352,P = 0.006),and the serum levels of THSD7A-AB were positively correlated with urinary protein and total cholesterol(r=0.289,P=0.035;r=0.352,P=0.006),negatively correlated with albumin(r=0.465,P<0.02),not associated with Scr(r = 0.785,P = 0.452),see Table 3.Conclusion:1 It is confirmed that THSD7 A is associated with the occurrence of IMN and is one of the latter pathogenic antigens.2 Detection of serum THSD7 A antibodies in patients with MN can assess the condition and understand the prognosis. |
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