الفهرس | Only 14 pages are availabe for public view |
Abstract Liver fibrosis results from a wound-healing response to chronic injury, which leads to excessive matrix, or scar deposition. This scar tissue leading to progressive liver damage and cirrhosis (the end stage of fibrosis), complicated by liver failure, portal hypertension and/or hepatocellular carcinoma (HCC). Genome-wide analysis of abnormal gene expression showed transcripts deregulation differences among normal, mild and severe fibrosis during HCC development with identification of novel serum markers for its early stage. Recent studies suggest that genetic markers may be able to define exact stage of liver fibrosis. In this study, the transcriptional profiling of fibrosis related genes in HCV-chronically infected patients with different degrees of liver fibrosis has been examined. PCR array was used to examine the expression of 84 fibrosis related genes in peripheral blood mononuclear cells (PBMCs). The RNA from 30 treatment-naïve chronic HCV patients (17 F0-F2 and 13 F3-F4) and 6 healthy subjects were enrolled. Validation of the results were performed by custom real time PCR in 75 treatment-naïve chronic HCV patients (39 F0-F2 and 36 F3-F4) and 10 controls. from the first approach data (PCR array data), selected eleven genes were based on their fold change, physiological pathway and p value. In the second approach, custom real time PCR confirmed that expression of transforming growth factor β (TGF β1) gene at the early stage of fibrosis is highly regulated compared to control (fold differential regulation =2.555033, p= 0.035), whereas the TG- interacting factor (TGIF1) gene which is the inhibitory of TGF β1 is down regulated (fold differential regulation =-1.2, p= 0.041). At the late stage of fibrosis, TGF β1 gene showed no significant changes in fold regulation when compared with control, while the TGIF1 gene is sharply down regulated (fold differential regulation =-1.6,p= 0.01). Whereas the TGFβ2 gene is sharply down regulated in the late stage without statistically significant. Also there is increase in Matrix Metalloproteinase 9 (MMP9) fold regulation in the late stage than early stage when compared with control, but without statistically significant. Moreover, by custom real time PCR: TIMP1, TIMP2, IL1B, SP1, PDGFA and THBS1 genes showed no significant changes in fold regulation in early and late fibrotic patients when compared with control. While these results suggest that TGFβ1 and TGIF1 are possible candidates as biomarkers for the progression of HCVinduced liver fibrosis and/ or targets for therapeutic intervention. Furthermore, the circulating proteins of the extra cellular matrix (ECM) was evaluated. ELISA has showed that the level of secreted TIMP2, TIMP1 proteins were significantly higher in late fibrotic patients (**p=0.000, *p=0.034) respectively than its conc, in serum of early fibrotic patients (F0-F2). On the other hand, MMP9 protein was significantly higher in early fibrotic patients than its conc, in serum of late fibrotic patients (*p=0.022). Moreover, the non-parametric Kruskal-Wallis test showed significant difference for TIMP2, MMP9 proteins levels among different fibrotic stages p=0.001, 0.042, respectively, while TIMP1 was not significant p=0.1.Receiver Operating characteristic (ROC) analyses curve suggested that TGFβ1 mRNA, TIMP2 and TIMP1proteins (AUC=0.694, 0.777 and 0.659 respectively) are possible candidates as biomarkers for the progression of HCV-induced liver fibrosis and/ or adjuvants for therapeutic intervention to stop failing of the clinical course of HCV chronic infection in patients. Keywords: HCV, gene expression, liver fibrosis, RT-PCR. |