Toxic Effects And Mechanisms Of Advanced Glycation End Products On Human Renal Tubular Epithelial Cells

Objective:To investigate the cytotoxicity of human renal tubular epithelial cells(HKC) induced by advanced glycation end products(AGEs) and explore the possible mechanisms.Methods:1.AGEs were prepared in vitro by incubating glucose with bovine serum albumin(BSA). The fluorescent intensity of AGEs is measured by spectrofluorophotometer.2.The proliferative inhibition of HKC induced by AGEs at different concentrations and different time was accessed by using MTT assay.Cell apoptosis was analyzed by flow cytometry and DNA agarose gel electrophoresis.3.The expression of IL-1β,TNF-αand HMGB1 mRNA in HKC cells induced by AGEs were determined by RT-PCR..4.The generation of reactive oxygen species(ROS) was determined by spectrofluorophoto-meter. The activity of SOD was detected by UV spectrophotometer.Results:1.Compared with control group(nontreated with AGEs),AGEs significantly inhibited proliferation of HKC,which showed good time- and dose-dependent fashions. 2.AGEs were also found to cause apoptosis in HKC.The flow cytometry analysis showed that AGEs increased the proportion of apoptotic cells in a dose-dependent manner.The DNA Ladder can be observed when the HKC was treated by AGEs.3.RT-PCR analysis demonstrated that the expression of IL-1β,TNF-αand HMGB1 mRNA in HKC cells were up-regulated by AGEs.4.The generation of ROS and the activity of SOD was elevated with AGES concentration increasing.Conlusions:1.AGEs can efficiently inhibit HKC proliferation,these growth inhibitions of HKC induced by the AGEs showed good time- and dose- dependent fashions.2.Apoptosis of HKC can be significantly promoted by AGEs.3.The cytotoxicity of HKC induced by AGEs may be related to the up-regulated expression of IL-1β,TNF-αand HMGB1 mRNA.4.The generation of ROS and the activity of SOD increased when the HKC was treated by AGEs.It suggested that the cytotoxicity of HKC induced by AGEs may be connected with oxidative stress.