AIM: To investigate the changes of retinal nerve fiber layer (RNFL) among normal individuals, diabetic patients without diabetic retinopathy (NDR) and non-proliferative diabetic retinopathy (NPDR), and explore the possible risk factors of early diabetic retinopathy (DR). METHODS: In this cross-sectional study, 107 participants were divided in three groups. Totally 31 normal individuals (control group), 40 diabetic patients without DR (NDR group) and 36 patients with NPDR (NPDR group) were included. Optical coherence tomography (OCT) was used to detect RNFL thickness and other optic disc parameters among different groups. The potential association between RNFL loss and systemic risk factors were assessed for DR, including diabetes duration, body mass index (BMI), hemoglobin A1c (HbA1c), serum lipids, and blood pressure. RESULTS: The average and each quadrant RNFL thickness were thinner in NPDR group compared to control group of the right (P=0.00, P=0.01, P=0.01, P=0.02, P=0.04) and left eyes (P=0.00, P=0.00, P=0.00, P=0.03, P=0.04). The average, superior and inferior RNFL thickness were thinner in NDR group compared to the NPDR group of the right (P=0.00, P=0.02, P=0.03) and left eyes (P=0.00, P=0.00, P=0.01). Diabetic duration was negatively correlated with the superior, inferior, and average RNFL thickness of the right (r=-0.385, P=0.001; r=-0.366, P=0.001; r=-0.503, P=0.000) and left eyes (r=-0.271, P=0.018; r=0.278, P=0.015; r=-0.260, P=0.023). HbA1c was negatively correlated with the superior, inferior, and average RNFL thickness of the right (r=-0.316 P=0.005; r=-0.414, P=0.000; r=-0.418, P=0.000) and left eyes (r= -0.367, P=0.001; r=-0.250, P=0.030; r=-0.393, P=0.000). Systolic pressure was negatively correlated with the inferior and average RNFL thickness of the right eye (r=-0.402, P=0.000; r=-0.371, P=0.001) and was negatively correlated with the superior and average RNFL thickness of the left eye (r=-0.264, P=0.021; r=-0.233, P=0.043). CONCLUSION: RNFL loss, especially in the superior and inferior quadrants, may be the earliest structural change of the retina in diabetic patients, and is also associated with diabetic duration, HbA1c, and systolic pressure.