Background and Objective: Reports show that type 2 diabetic (T2D) patients have changes in red blood cell (RBC) indices as well as decreased erythropoietin (EPO) levels and endothelial nitric oxide synthase (eNOS) activity. Some abnormalities have been reported to develop during the pre-diabetes stage. However, information on haemolytic changes during the progression of pre-diabetic stage has not yet been reported. Therefore, this study was sought to characterize the changes in RBC indices and concentration of eNOS and EPO in the pre-diabetic stage. Materials and Methods: Pre-diabetes was induced using a high-fat high-carbohydrate diet for 20 weeks. Rats were then divided into non-diabetic and prediabetic rats (n = 6 in each group). The pre-diabetes progressed to T2D over an additional 12-week experimental period. RBC indices were measured at the end of 20 and 32 week. Following sacrificed after 32 weeks, blood was collected for eNOS and EPO measurements. Results: The results showed significant increases in RBCs, hemoglobin (HGB), hematocrit (HCT) in the prediabetic group as compared to the non-prediabetic group. We further observed a significant decrease in white blood cell (WBC) concentration, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) and red cell distribution width (RDW) in the prediabetic group as compared to the non-prediabetic group. Conclusion: Changes in blood cells indices indicate various haemolytic changes in RBCs morphology during progression towards overt T2D. However, EPO upregulation indicates the production of new RBCs, while decrease in eNOS activity indicated decrease in bioavailability of nitric oxide (NO).