雷帕霉素对高糖诱导人晶状体上皮细胞氧化应激的保护作用

Rapamycin protects human lens epithelial cells against oxidative stress induced by high glucose

目的探讨雷帕霉素对高糖诱导的人晶状体上皮细胞(human lens epithelial cells,HLECs)氧化应激的影响。方法建立高糖诱导的HLECs氧化损伤模型,用不同浓度(10 nmol·L-1、100 nmol·L-1)雷帕霉素干预,CCK-8比色法检测细胞存活率,倒置显微镜观察细胞形态并拍照,H2DCFDA检测细胞内活性氧水平,Western Blotting检测各组凋亡相关蛋白Bcl-2及Bax的表达情况。结果 CCK-8检测结果显示,用10 nmol·L-1、100 nmol·L-1雷帕霉素处理后,HLECs存活率分别提高到62.86%±3.45%和77.47%±4.21%,与高糖组(42.32%±3.10%)比较差异均有统计学意义(均为P<0.05);H2DCFDA荧光探针染色结果显示,雷帕霉素处理后HLECs内活性氧生成量下降,氧化损伤细胞减少;此外,雷帕霉素可以抑制高糖诱导的HLECs内凋亡蛋白Bax、Bcl-2的表达,抑制效应与剂量正相关。结论雷帕霉素可以抑制高糖诱导的HLECs氧化损伤及凋亡的发生,对糖尿病性白内障具有预防作用。

ObjectiveTo investigate the effects of rapamycin on human lens epithelial cells(HLECs)against high glucose induced oxidative stress.MethodsA oxidative damage model of HLECs induced by high glucose was established,and intervention with different concentrations of rapamycin for the cells was performed for detecting the survival rate by CCK 8 assay,observing cell morphology by inverted microscope,and measuring the level of intracellular reactive oxygen species(ROS)by H2DCFDA staining,as well as determining the expression of apoptosis related Bcl 2 and Bax proteins by Western blot in all groups.ResultsCCK 8 results showed that the survival rate of HLECs was increased to 62.00%±3.74%and 79.57%±5.26%after treated with 10 nmol·L-1 and 100 nmol·L-1 rapamycin,and the difference was statistically significant when compared with the high glucose group(42.32%±3.10%)(all P<0.05);H2DCFDA fluorescent probe staining demonstrated that rapamycin could suppress ROS generation in HLECs and alleviate oxidative damage to cells;besides,rapamycin could downregulate the expression of Bcl 2 and Bax protein in HLECs and the inhibitory effects were positively correlated with the concentration of rapamycin.ConclusionRapamycin can inhibit high glucose induced oxidative damage and apoptosis in human HLECs,which provides a prevention effect for diabetic cataract.