Background: Hepatitis C Virus (HCV) infection is a progressive disease that may result in chronic hepatitis, fibrosis and cirrhosis. Assessment of liver fibrosis is an essential factor in the management of chronic HCV. Objective: To evaluate plasma soluble Urokinase Plasminogen Activator Receptor (sU-PAR) and interleukin-34 (IL-34) as serological markers of liver fibrosis in patients with chronic HCV. Methods: This case-control study enrolled 60 chronic HCV patients who were subdivided into three groups of mild, moderate and severe hepatic fibrosis depending on Fibrosis-4 score (FIB-4). Patients were compared with 20 age and sex-matched controls. Plasma sUPAR and IL-34 levels were measured by Enzyme Linked Immunosorbent Assay (ELISA). Results: Plasma sUPAR and IL-34 were significantly increased in HCV patients when compared with controls, and their increase was positively correlated with the progression of hepatic fibrosis. Plasma sUPAR and IL-34 positively correlated with Aspartate Aminotransferase (AST) and Alanine Aminotransferase (ALT), and negatively correlated with hemoglobin concentration and platelet count. The output data of Receiver Operating Characteristic (ROC) curve to differentiate patients from controls revealed that sU-PAR at cut-off > 186.2 ng/L and Area Under Curve (AUC) of 0.944 had (85%) sensitivity and (100%) specificity, and IL-34 at cut off > 16.4 ng/L and AUC of 0.942 had (75%) sensitivity and (100%) specificity. The output data of ROC curve to differentiate severe from mild to moderate hepatic fibrosis patients revealed that sUPAR at cut-off > 510 ng/L and AUC of 0.