Background Renal function assessment in T2DM individuals is very important, as diabetic nephropathy (DN) seems to be the major cause of chronic kidney disease (CKD) that may finally lead to End Stage Renal Disease (ESRD) in those patients. Clearly, DN pathogenesis is multifactorial and different proteins, genes and environmental factors can contribute to the onset of the disease. Therefore, we need to assess sensitive and specific biomarkers which can predict susceptibility of development of kidney disease in those patients. Methodology: The current study was performed on 108 T2DM patients and 85 normal controls (NC). Biochemical parameters as blood glucose, HbA1c and serum Cystatin-C were assessed. Genetic determination of both ACE I/D and ACE G2350A was done for all participants. Also, urinary hemeoxygenase (HO-1) was evaluated as a new biomarker of kidney disease. Results showed significant rise in serum cystatin-c and urinary hemeoxygenase levels in diabetic groups compared with (NC) group. In addition, GG genotype of ACE G2350A gene in diabetic group was associated with rise in serum cystatin-c and urinary heme oxygenase compared with (NC) group. Mutant AA genotype demonstrated increase in urinary HO-1. DD polymorphism was associated with rise in serum creatinine and Cyst-C in diabetic group. Positive correlation was seen between duration of diabetes and serum cystatin-c and between serum glucose and urinary (HO-1) in diabetic group. Conclusion Early detection of nephropathy in T2DM patients can enable adequate treatment and improve their outcomes. Our study indicated an association of GG genotype of ACE G2350A in conjugation with DD polymorphism of ACE I/D with serum cystatin-c as early predictor of tubular injury in T2DM diabetic.