ABSTRACT
Objective To investigate the effects of rapamycin on human lens epithelial cells (HLECs) against high glucose-induced oxidative stress.Methods A 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.Results CCK-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.Conclusion Rapamycin can inhibit high glucose-induced oxidative damage and apoptosis m human HLECs,which provides a prevention effect for diabetic cataract.
ABSTRACT
Abstract?AIM: To investigate the effect of epigallocatechin-3-gallate ( EGCG ) against oxidative stress induced by high glucose in human lens epithelium ( HLE) cells.? METHODS: The HLE cell oxidative damage model induced by high concentration glucose was established, and was intervented with different concentrations of EGCG. Cell viability was determined by MTT assay, cell morphology was investigated by convert microscope, cells apoptosis was assayed by flow cytometry with Hoechst-PI staining. Moreover, the levels of super oxide dismutase ( SOD) , glutathione peroxidase ( GSH-Px) and malondialdehyde ( MDA) in supernatant were also tested after different treatment either with high concentration glucose or with different concentrations of EGCG.?RESULTS: MTT results showed that HLE cells activity increased to 50. 33%± 3. 52% and 63. 33%± 4. 63% after treated with 10 μmol/L and 100 μmol/L EGCG respectively, the difference was statistically significant compared with oxidative injury group(32. 67%±3. 10%)(P<0. 05 ); HLE cells maintained better morphology intervented with EGCG under high glucose conditions, the number of apoptotic cells reduced, SOD and GSH-Px level within HLE cells increased and MDA levels decreased.?CONCLUSION:EGCG plays its strong antioxidant effect by increasing SOD, GSH-Px content and decreasing MDA content in cells, therefore provides a reliable experimental basis for the search for effective prevention and treatment of cataract drug.