Experiment Research Of Endoplasmic Reticulum Stress On Oxidative Injury Of Human Lens Epithelial Cells

Part 1 The importance to the measurement of GRP78 , Caspase-12 and the apoptosis of LEC post of the oxidative injuryObjective Judging from the variance of GRP78, Caspase-12 and the apoptosis of LEC to discuss the regularity of those in LEC post oxidative injury and the relationship with apoptosis of LEC.Methods Cultured LEC (SRA01/04) were divided into two groups: control group and oxidative injured group which was treated with 50umol/L H₂O₂ for 24 hours. Cells apoptosis and the mRNA expression of GRP78 and Caspase-12 were analyzed. The apoptosis of LEC was observed by TUNEL techniques. RT-PCR was used to determine the mRNA expression of GRP78 and Caspase-12 in LEC. All the results were analyzed by the statistics method.Results1. The apoptotic rate of LEC: the apoptotic rate of LEC in control group was 5.5%, and the result in oxidative injured group was 49.3%. There was significant difference between the control group and oxidative injured group (P<0.05) .2. The mRNA expression of the GRP78: the relative optical density of GRP78 in control group was 1.164±0.007, and the result in oxidative injured group was 1.306±0.007. There was significant difference between the control group and oxidative injurec group (P<0.05) .3. The mRNA expression of the Caspase-12: the relative optical density of Caspase-12 in control group was 0.201±0.021, and the result in oxidative injured group was 1.273±0.039. There was significant difference between the control group and oxidative injurec group (P<0.05) .Conclusions As the marker of ERS, the mRNA expressions of GRP78 and Caspase-12 increased, and also the rate of apoptosis rised during the LEC was injured by oxidative factor. These showed the ERS participated in the apoptosis process of oxidative injured LEC.Part 2 Experimental research on the expression of GRP78 and Caspase-12 in 2-DG pretreated oxidative injured LECObjective On purpose of surveying the effect of GRP78 and Caspase-12 induced by 2-DG pretreatment on LEC under oxidative injury, the mRNA expressions of GRP78 and Caspase-12, and also the apoptosis of LEC post ERS preliminary adaptation were observed.Methods Cultured LEC (SRA01/04) were divided into four groups: Group A (control group), Group B(oxidative injury group) ,Group C (4mmol/L2-DG pretreated group) and Group D (8mmol/L 2-DG pretreated group). Cells apoptotic rate and the mRNA expressions of GRP78 and Caspase-12 were analyzed. The apoptotic rate of LEC was observed by TUNEL techniques. RT-PCR was used to determine the mRNA expressions of GRP78 and Caspase-12 in LEC.Results1. The apoptotic rate of LEC: the apoptotic rate of LEC in Group A, B, C and D were 5.5%, 49.3%, 24.6% and 18.75%. There was significant difference among four groups (P<0.05) .2. The mRNA expression of the GRP78: the relative optical density of GRP78 in Group A, B, C and D were 1.164±0.007, 1.306±0.007. 1.515±0.004 and 1.535±0.004.There was significant difference among the four groups (P<0.05)3. The mRNA expression of the Caspase-12: the relative optical density of Caspase-12 in Group A, B, C and D were 0.201±0.021, 1.273±0.039. 0.783±0.023 and 0.721±0.019. There was significant difference among the four groups (P<0.05)Conclusions To play a protective role, the increased expression of GRP78 induced by 2-DG pretreatment might inhibit the apoptosis of LEC by reducing the Caspase-12.