| Purpose:The world health organization(WHO) reports that about2billion people had HBV infection in the world. Patients with chronic hepatitis B may develop into liver fibrosis, cirrhosis, even hepatocellular carcinoma(HCC). Most of liver fibrosis is caused by chronic hepatitis B.It is helpful for the diagnosis and clinical treatment to judgment the development of liver fibrosis. For the diagnosis of liver fibrosis and cirrhosis, now, Liver biopsy pathologic examination is still the gold standard. Since it has traumatic to patients, the discomfort and the potential complications may occur. And some safe and convenient testing is needed to study.With the development of the technology of proteomics, the subject has it’s unique advantage in the discovering of specific Biomarker for desease. And PBMC is the important immune cell, and it is involved in the development process of many diseases. Therefore, our study is designed to explore the differences of protein expression in PBMC with different stages of liver fibrosis by proteomics route,aims to find specific biomarkers related to liver fibrosis stages for the stage evaluation and guidance for the diagnosis, treatment or prognosis of liver fibrosis. Methods:Patients with chronic hepatitis B liver fibrosis were collected in Shanghai Public Health Clinical Center Affiliated to Fudan University in2009.3-2011.2.48samples were divided into Sl group (n=24), S4group (n=24) according to liver biopsy diagnostic stages of hepatic fibrosis. All patients enrolled were aged34to63. No statistically significant difference was found in age(P=0.780).PBMC was separated by lymphocyte separation medium samples. The concentration of protein extracted from PBMC was detected by the method named Bradford.The protein were separated by two-dimensional electrophoresis, and analyzed by ImageMaster2D Platinum6.0software to find differentially expressed proteins related to liver fibrosis stages.After digested by trypsin, the differential proteins were identified by online reversed-phase nano-flow liquid chromatography combined with electrospray ionization ion trap mass spectrometry. Ascertain the protein information by compared to data bank, and to find out it’s biological significances by consulting literatures.Results:protein was efficiently separated, and1057±134spots were detected in Sl group, and1052±130spots in Sl group. The matched spots in Sl group are727±161(61.5%), and in Sl group are716±139(58.1%).12differentially expressed protein spots were acquired.8protein spots of them were up-regulated otherwise4down-regulated. All the12different protein spots were identified accurately. The identified proteins are related to cell movement, cell adhesion and transcription etc.After consulting literatures, we found out that the Moesin, Transgelin-2and HSP70may be related to liver fibrosis stages, and may provide a new area for the study of the pathogenesis mechanism of liver fibrosis. Conclusions:We apply proteomics techniques in screening and identifying some proteins from PBMC, and the proteins may be related to liver fibrosis progress, and may make sense for the study of the development of liver fibrosis. Proteomics route on PBMC may make some contribution to seeking and foundation of biological markers related to liver fibrosis stages. |
