| Objective: To observe the changes in the expression of P66 shc in retinal tissue by using STZ induced diabetic rat model,and to explore the relationship between retinal tissue injury and retinal tissue injury.Methods: the experiment was divided into the experimental group and the normal control group,with 10 SD rats in each group,with a weight of 160-200 grams.The diabetic rat model of SD was established by intraperitoneal injection of STZ30mg/Kg(the control group was injected with equal amount of PBS).The fasting blood glucose greater than16.7 mmol/L were selected as the experimental group.In the two groups,5 rats were taken each of the rats in February and March respectively.They were killed and retina tissue was taken.Electron microscopy was used to observe the retinal tissue structure changes,changes in localization and expression of p66 Shc in retinal tissue was detected by immunofluorescence;Western blot(Western Blot,WB),the content of p66 Shc protein in rat retina was detected;expression of RT-PCR in retinal p66 Shc mRNA.Results: SD rats 72 hours after intraperitoneal injection of STZ,fasting blood glucose greater than 16.7 mmol/L,showed that the diabetic rats model was successful.The blood sugar of the rats in the experimental group was obviously increased,and the symptoms of polypodities,polypodities and polyuria were obvious,and the weight of the rats was lighter than the control group.In February and March when the rats were sacrificed,retinal tissue detection,electron microscopy showed: normal rat retinal structure is clear,the rods and cones arranged orderly,cytoplasm showed a large amount of endoplasmic reticulum and mitochondria,and ganglion cells of varying size,uniform distribution,organelles were normal,the structure of normal distribution uniform.The capillary endothelial cells in the diabetic control group were edema,and the mitochondria were swollen.The space of the outer segment of the cell was enlarged and arranged in disorder.In the inner segment,the mitochondria were edema,irregular arrangement,vacuolization appeared in the cells,the perinuclear space increased,arranged in disorder,and the cells became loose and necrotic and arranged loosely.The lesions were more severe in 3 months than 2 months.Immunofluorescence showed that the normal group and experimental group rats whole retina structure has the expression of p66 Shc,the endoplasmic reticulum layer and nerve cell layer and nerve fiber layer,inner limiting membrane expression of high expression increased from the choroid to vitreous direction,mainly located in the neural retina of experimental group,the red fluorescence in February and in March were significantly higher than the control group,March is more obvious,the results of semi quantitative analysis showed that the expression of p66 Shc in the retina tissue of diabetic rats was changed compared with that of the normal group,showing a significant trend of enhancement,and the difference was statistically significant.(P < 0.05 group,February,March group P < 0.01).RT-PCR results showed that the expression of p66 Shc mRNA in theretina tissue of the experimental group increased significantly in February and March,and it was more obvious in March than in the control group.The difference was statistically significant(P < 0.05,P < 0.01).Results of Western Blot analysis: the expression of p66 Shc protein in the retina of experimental group increased significantly,compared with the control group,there was significant difference between February and March,and the difference was statistically significant in March group(P < 0.05,P < 0.01).Conclusion: the expression of p66 Shc in retina tissue of diabetic retinopathy rats increased significantly,suggesting that it may play an important role in the process of diabetic retinopathy. |
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