Abstract:AIM: To observe the effects of advanced glycation end-products(AGEs)on thioltransferase(TTase)expression and activity in human lens epithelial cells.
METHODS: Human lens epithelial cells B3(HLE B3)were treated with 1.5mg/mL AGEs-BSA as the experimental groups cultured by fetal bovine serum of volume fraction 10% dulbecco modified eagle medium(DMEM)and bovine serum albumin(BSA)was added at the same concentrations as the negative control. The level of reactive oxygen species(ROS)was evaluated. Cells were collected at 0, 1, 2, 3, 4d and total RNA or protein was extracted. TTase mRNA levels were detected by qRT-RCR. TTase expression was detected by Western blot and its activity was measured.
RESULTS: Compared with the control group,AGEs-BSA up-regulated the expression of ROS(P<0.01), ROS content increased in a time-dependent manner. BSA had no effects on ROS expression. The expression of TTase increased after treatment with AGEs-BSA for 1d, peaked at 2d(nearly 5.06-fold increase, P<0.01), then decreased gradually. No change was observed between BSA and control group(P>0.05). Similarly, TTase activity peaked at 3d(nearly 2.01-fold increase, P<0.01). Western blot test found that TTase protein expression was increased gradually, starting from the 3d TTase expression was reflected that there was statistically significant difference compared with control group(P<0.05).
CONCLUSION:AGEs-BSA significantly increases the production of ROS in human lens epithelial cells, and it then induces the oxidative stress which may promote the expression of TTase and enhances the activity of TTase.