Aim To explore the effect of advanced glycation end products (AGE) on injuring of rat renal microvascular endothelial cells (RMEC) and protective effect of probucol. Methods Microvascular endothelial cells isolated and cultured from rat renal were divided into 3 groups: normal control group, AGE group and probucol group. The levels of malondialdehyde (MDA) and nitric oxide (NO) were determined by the assay TBA and nitrate reductase method respectively. The expression of endothelial nitric oxide synthase (eNOS) mRNA and nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase protein was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western-blot respectively. The intracellular disposition of nuclear factor kappa B (NF-κB) was observed by immunostaining microscopy. Results In AGE group, the level of MDA increased but the expression of NADPH oxidase protein and eNOS mRNA decreased compared with control. The level of NO in culture medium was increased after exposure to AGE-BSA for 30 min and 6 h,but after 12 h, NO level was decreased. MDA level and the expression of NADPH oxidase protein were decreased but the expression of eNOS mRNA and NO level were upregulated by probucol with a dose-dependent effect in the concentrations of 10, 20, 50, 100 μmol/L. The localization of NF-κB shifted from cytoplasm to nucleus after microvascular endothelial cells were exposed to AGE-BSA. After exposed for 30 min, the expression in nucleus reached the peak and then maintained high level. Conclusion AGE can cause the injury and dysfunction of RMEC via the following possible mechanisms: ①inducing lipid preoxidation; ②activating NADPH oxidase and increasing the production of reactive oxygen species (ROS); ③abnormal expression of eNOS mRNA and further affecting NO production; ④activatin of NF-κB. Probucol can protect microvascular entothelial cells probably via antagonisting the lipid preoxidation, activating of NF-κB and NADPH oxidase,and increasing the AGE-induced abnormal expression of eNOS caused by AGE.