Objective To investigate the role of podocytes damage in the process of glomerular sclerosis by observing the clinical and pathological indices and the expression of nephrin and Wilms’ tumor-1 (WT-1) in the podocytes of puromycin aminonucleoside (PAN)-induced nephropathy model. Methods A single intravenous injection of PAN via tail vein of rats was carried out to establish nephropathy model, and those with positive urine?protein were identified as successful establishment. Biochemical and pathological parameters were analyzed in the model rats and normal controls at 4 and 8 d, and at 4, 14 and 20 weeks after the model establishment for the chronic renal injuries evaluation. After the rats were sacrificed at 4, 14 and 20 weeks, their kidneys were taken out, embedded in paraffin, and stained with periodic acid-Schiff (PAS) for determination of glomerulosclerosis index (GSI) and glomerular area (GA). Immunohistochemistry assay was performed to detect the expression of nephrin and WT-1. Results The PAN models were successfully established, showing typical nephritic syndrome in the early stage (day 8), and chronic glomerular nephritis in the later stage (week 20). The GSI was significantly higher in PAN group than in the normal controls at 4, 14 and 20 weeks after establishment (P＜0.05). The mean size of GA was significantly larger in PAN rats than in the normal controls at 14 weeks (P＜0.01), but was obviously smaller at 20 weeks (P＜0.01). Significantly positive correlation was found between GSI and GA at 20 weeks in the model group. The expression of nephrin was obviously decreased in PAN group at 4, 14 and 20 weeks (P＜0.05), so was that of WT-1 at 14 and 20 weeks (P＜0.001). Conclusion (1) GSI is increased, while GA is decreased along with the progression of nephritis, and there is a negative correlation between GSI and GA. (2) The expression of nephrin and WT-1 is reduced along with the progression of glomerular sclerosis, indicating the decrease of podocytes. Therefore, the podocyte damage is closely associated with glomerular sclerosis.