方法：将体外人晶状体上皮细胞用AGEs-BSA浓度为1.5mg/mL，胎牛血清体积分数为10%的DMEM培养液培养，同时设定相同浓度的BSA对照组及空白对照组，分别于0，1，2，3，4d收集细胞，测定晶状体上皮细胞内ROS含量、TTase活性，qRT-PCR检测TTase mRNA表达情况，Western blot检测TTase蛋白质表达。

结果：与对照组相比，AGEs-BSA干预后，细胞内ROS含量呈时间依赖性增高，差异有显著统计学意义(P<0.01)，BSA干预后ROS的表达与对照组无显著差异。AGEs可诱导TTase的 mRNA表达逐渐增高，2d时达到峰值，约为正常对照组的5.06倍(P<0.01)； 而BSA处理组和对照组TTase的mRNA表达差异无统计学意义(P>0.05)。与TTase的mRNA表达类似，TTase活性升高，在3d达到峰值，为正常对照组的2.01倍(P<0.01)。Western blot检测发现，TTase蛋白质表达逐渐增加，从3d开始TTase表达与对照组相比差异有统计学意义(P<0.05)。

结论：AGEs可能是通过诱导人晶状体上皮细胞发生氧化应激，致使TTase表达上调，活性增强。

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.