Primary Study On Urine Proteomics In Hepatitis B Virus Associated Glomerulonephritis

Objective:To analyse the polypeptide pattern in urine of patitents with hepatitis B virus associated glomerulonephritis,and compare with the urine protein of primary glomerulonephritis complicated with chronic hepatitis B and healthy people.Aim to find the protein which has a high sensitivity and specificity,and to discuss whether this echnology is suitable for clinical diagnosis.Methods:Morning first medistream urine samples from 3 healthy people and 9 patients were collected.In order to avoid bacterial growth and protein enzymolysis,stored samples at -80℃during 1h.After melting, every sample was filtered through 0.22μm D filtration membrane to remove impurities.Urine samples were frationated by acetone precipitation (the volume ratio was 4:1)at -20℃overnight.Then centrifugated at 4℃4000xg for 20 minutes,and obtained the precipitate which contained urine protein, then dried down in a vacuum centrifuge. The protein concentration of each sample was determined by Bradford assay. The protein was digested with trypsin overnight at 37℃.Then went to HPLC-ESI-MS /MS. The protein identification was searched in the database of ipi. human, v3.05with the help of MASCOT.Results:101 proteins were identificated from healthy people.There were 37 proteins commonly expressed.73 proteins were identificated from the urine of patients with primary glomerulonephritis complicated with chronic hepatitis B.There were 15 proteins commonly expressed.206 pro-teins were identificated from the urine of patients with hepatitis B virus associated glomerulonephritis. There were 19 proteins commonly expressed. Rejecting urine protein of healthy people and the patients with primary glomerulonephritis complicated with chronic hepatitis B from proteins commonly expressed in patients with hepatitis B virus associated glo-merulonephritis,9 proteins were left.They are ceruloplasmin,alpha-1-antichymotrypsin,Ig lambda chain C region,Ig gamma-2 chain C region,Ig alpha-1 chain C region,Ig alpha-2 chain C region,vitamin D-binding protein,prostaglandin D2 synthase,C9JAM6(putative uncharacterized pro-tein IGHA1).Conclusion:We used HPLC-ESI-MS/MS to analyse the polypeptide pattern in urine of patitents with hepatitis B virus associated glomerulo-nephritis,206 protein were identificated.Compare with the urine protein of healthy people and the patients with primary glomerulonephritis compli-cated with chronic hepatitis B,9 proteins specifically expressed in pati-tents with hepatitis B virus associated glomerulonephritis.Ceruloplasmin,alpha-1-antichymotrypsin,vitamin D-binding protein and prostaglandin D2 synthase may become biomarkers of hepatitis B virus associated glo-merulonephritis.