Hepatic fibrosis is an early event in cirrhosis in patients with HBV infection. The gold standards for the diagnosis of hepatic fibrosis are pathological staging and classification. However, biopsy of hepatic fibrosis in patients is lim ited by sampling error, poor compliance of patients, and difficulty in drawing the materials from live tissue. Methods selleck chemicals llc which will achieve early diagnosis of hepatic fi brosis are still unknown. The use of serum biomarkers for diagnosis of hepatic fibrosis has many advantages including Inhibitors,Modulators,Libraries being noninva sive, quick to acquire data, and provides confirmation of hepatic fibrosis in HBV infected patients faster. However, there are no suitable serum biomarkers known that can serve as a reliable diagnostic biomarker for hep atic fibrosis.
Proteomics is an effective method to obtain high flux protein data useful for identifying biomarkers. Inhibitors,Modulators,Libraries In this study, in an effort to identify a serum biomarker of hepatic fibrosis, we used 2 DE and MALDI TOF MS to compare proteins which were differentially expressed in the serum of patients with hepatic fibrosis compared to HBV carriers. We identified two proteins, enolase 1 and Thrombospondin 1, which were differentially expressed and were chosen for further study. Our study suggests enolase 1 and TSP 1 play important roles in the development of hep atic fibrosis. Materials and methods Serum samples Serum samples Inhibitors,Modulators,Libraries include 4 from patients with HBV hepatic fibrosis and another 4 from HBV carriers. All samples were from the First Xiangya Hospital of Central South University during the period of 2008. 1 to 2010.
12. All patients with HBV Inhibitors,Modulators,Libraries hepatic fibrosis were confirmed by pathological biopsy by two pathologists. The diagnostic criteria to classify a patient as an HBV carrier were as follows HBsAg, HBV DNA, HBeAg or anti HBeAb, and with normal level of ALT and AST, and no abnormal histological biopsy. All serum samples were drawn using a K3 EDTA anticoagulation tube and stored at 4 C in the freezer after centrifuging at 2500 rpm 15 min The blood plasma of the upper layer was taken using suction and the samples were stored at 80 C. Reagents Micro Bio Spin column and protein assay reagents were from Bio Rad. Mouse anti human GAPDH antibody was purchased from Sigma Co. Ltd, Mouse anti Inhibitors,Modulators,Libraries human enolase 1, TSP 1 and secondary antibody were purchased from Santa Cruz Co. Ltd.
ProteoExtract sellckchem Subcellular Proteome Extraction Kit was purchased from MERCK Co. Ltd. Methods Serum sample preparation All serum samples were transported on ice and centrifuged at 3000 g for 15 min at 4 C. The superna tants were stored at 80 C until further analysis. Serum samples from two patients with HBV hepatic fibrosis and two HBV carriers were used for the proteomics study. The serum samples from the other four patients with HBV hepatic fibrosis and four HBV carriers were used for the western blotting study.