Association between lipids and renal disease has been reported recently. Its pathogenic mechanisms remain unknown. The aims of this study were to establish: (1) if a cholesterol-rich diet, alone or associated with nephrectomy, produces nephropathy; and (2) if a treatment with omega-3 polyunsaturated fatty acids (PUFA) reduces glomerulosclerotic lesions. Sixty Sprague-Dawley rats were randomized in two different groups: (A) sham operated rats and (B) uninephrectomized rats. Rats in both groups were divided into three subgroups (A1-3, B1-3) according to the diet they were fed: normal chow diet, cholesterol-rich diet (4.5%) or cholesterol-rich diet supplemented with omega-3 PUFA. Twenty weeks later, serum creatinine, creatinine clearance, serum cholesterol, triglycerides, albumin, proteinuria, mesangial cell score and focal glomerulosclerosis were assessed. Results showed that a cholesterol-rich diet significantly increased serum cholesterol, proteinuria and glomerular lesions and decreased creatinine clearance, especially in nephrectomized rats. Glomerular lesions, serum cholesterol and proteinuria ameliorated when cholesterol-rich diet was supplemented with PUFA. Hypertension was noticed only in nephrectomized rats following a normal chow diet. Simple correlation analysis showed that glomerulosclerosis correlated with renal weight, blood creatinine, cholesterol and proteinuria. In spite of some significant differences in urinary prostaglandins, no correlation with glomerular lesions was found. Multiple logistic regression analysis showed that cholesterol and proteinuria were independent risk factors for induction of glomerular sclerosis. In conclusion, a diet rich in cholesterol induces glomerulosclerosis, especially if it is associated with unilateral nephrectomy. Omega-3 PUFA administration reduces serum cholesterol, proteinuria and glomerular injury.