The inhibitory effect of decorin on apoptosis of human lens epithelial cells under high glucose condition and its mechanism

Objective

To investigate the effects of decorin (DCN) on apoptosis and oxidative stress in human lens epithelial cells (LECs) under high glucose condition.

Methods

HLE-B3 cells were cultured in vitro and the effect of DCN with different concentrations on HLE-B3 viability was determined by using cell counting kit-8 (CCK-8). The cultured cells were divided into normal control group, DCN group, high glucose group and DCN + high glucose group.Flow cytometry was used to detect the apoptosis rate and the expression of reactive oxygen species (ROS) in the cells.Microplate spectrophotometer was used to measure total superoxide dismutase (SOD) enzyme activity and the radio of glutathione (GSH)/glutathione disulfide (GSSG). Western blot was used to detect the expressions of bax and bcl-2 proteins.

Results

HLE-B3 cells were spindle shaped, with centered and clearly visible nuclei and neatly cell arrangment.According to CCK-8 method, survival rates of HLE-B3 in all groups were more than 90%.Different concentrations of DCN showed no significant effect on HLE-B3 survival rate (all at P>0.05). After 48 hours of cell culture, the apoptosis rate of high glucose group was significantly higher than that of normal control group, and the apoptosis rate of DCN+ high glucose group was significantly lower than that of high glucose group (both at P=0.000). The mean fluorescence intensity of intracellular ROS in the high-glucose group was significantly higher than that in the normal control group, and the mean fluorescence intensity of ROS in the DCN group was significantly higher than that in the high glucose group (both at P=0.000). The total SOD activity in the high glucose group was significantly lower than that in the normal control group and DCN group (P=0.007, 0.004). The GSH/GSSG ratio of the high-glucose group was significantly lower than that of the normal control group and DCN group (both at P=0.000).

Conclusions

DCN can inhibit the apoptosis and oxidative stress of HLE-B3 under high glucose, which provides the basis for the treatment of diabetic cataract.

Decorin; Human lens epithelial cell; High glucose condition; Oxidative stress; Apoptosis; Inhibitory effect; Mechanism