Cytotoxicity Of Advanced Glycation End Products And Their Functional Effects On Human Heptical Cell Line LO2

Objective:To investigate the cytotoxicity and the functional influences of advanced glycation end products(AGEs) on human heptical cell line LO2.Methods:1.AGEs were prepared in vitro by incubating glucose with bovine serum albumin(BSA). The content of AGEs was measured by using spectrofluorophotometer.2.Proliferative inhibitions of LO2 cells by AGEs at different concentrations and different time courses were accessed by using MTT assay.The number of cell divisions was observed by Fluorescence microscopy.Apoptosis in LO2 cells was analyzed by flow cytometry.3.Changes in the membrane surface of LO2 cells induced by AGEs were observed by atomic force microscopy.4.The activity changes of cellular aspartic transaminase caused by AGEs were determined by the clinical laboratory of the Oversea-Chinese Hospital,the first affiliated hospital of jinan university.5.The expression levels of IL-1β,TNF-α,HMGB1 and PKLR(Pyruvate kinase of liver and RBC) mRNA in LO2 cells treated with/without AGEs were determined by RT-PCR.Results:1.When compared with the control group(treated without AGEs),AGEs-treated groups showed significant inhibition of proliferation in LO2 cells,with good time-and dose-dependent fashions.2.AGEs were also found to cause apoptosis in LO2 cells.Flow cytometry analysis showed that AGEs increased the proportion of apoptotic cells in a time-and dose-dependent manner.3.Fluorescence microscopy showed that the number of cell divisions was reduced by AGEs.4.Atomic force microscopy analysis showed significant morphological changes in LO2 cells. The cell membrane surface became rough when treated with AGEs.5.Activity of aspartic transaminase in LO2 cells was found to be decreased by AGEs.6.RT-PCR analysis demonstrated that the expression levels of IL-1β,TNF-αand HMGB1 mRNA in LO2 cells were up-regulated by AGEs,while the expression of PKLR mRNA was down-regulated by AGEs.Conlusions:1.AGEs significantly inhibit LO2 proliferation,and the growth inhibition of LO2 by AGEs is well time-and dose-dependent.2.Apoptosis of LO2 cells may be induced by AGEs.3.The cytotoxicity in LO2 cells caused by AGEs may be related to the up-regulated expression of IL-1β,TNF-αand HMGB1.4.AGEs cause rough surface changes in LO2 cells,and damage the membrane structures of LO2 cells.5.AGEs decrease the activity of aspartic transaminase in LO2 cells,and down-regulate the expression of PKLR mRNA.