Effect Of AGEs On The Oxidative Stress And Activity Of NF-κB In Rat Lacrimal Gland Epithelium Cells

Objective:To observe the effect of advanced glycati on end products(AGEs)on the oxidative stress and activity of NF-κB in cultured rat lacrimal gland(LG) epithelium cells.Methods:1. To culture the primary rat LG epithelium cells by enzyme digesting2. To prepare AGEs-BSA by incubating both bovine serum albumin and sugar3. To exposed LG epithelium cells to AGEs(12.5,25,50,100,200μg/ml) for 48 hours,then chose the sensitive 100μg/ml AGEs to affect LG epithelium cells for 12,24,48 hours.The amounts of maleic dialdehyde and the activity of superoxide dismutase in LG epithelium cells were examined by spectrophotometry.The activity of NF-κB was examined by western blot.Result:1. The purity of rat LG epithelium cells is over 85%.2.Fluorescence spectrophotometry assess the fluorescence value of AGEs-BSA: zero adjustment with PBS,excitation wave length 390 nm, emission wave length 450 nm, peak narrow 5 nm.3. AGEs decreased the activity of superoxide dismutase,whereas increased the amounts of malondialdehvde maleic dialdehyde and the activity of NF-κB of cultured LG epithelium cells in a dose-dependent and time-dependent manner(P<0.01).Conclusion:1. The result shows that the isolation and culture of rat LG epithehlial cells by enzyme digesting is successful.2. AGEs promote the expression of NF-κB p65 in rat LG epithelium cells in a dose-dependent and time-dependent manner during some certain scope.The mechanism is probably associated with increased oxygen free radical produced by combination between AGEs and RAGE. It is conclude that AGEs probably induce or aggravate inflammation of LG in vivo which lead to diabetic and age-related LG pathological changes.