Difference Analysis Of Proteomics Between Diabetic Cataract And Normal SD Rats' Lens

Diabetic cataract is a common diabetes complications, and was second in the diabetic eye disease. Its pathogenesis have many kinds,such as:Osmotic pressure theory, Protein glycosyl theory, Oxidative stress theory. Because of the lens is the highest biology organization content of protein and Very suitable for application of proteomics approach to study it.In recent years, with the establishment and development of proteomics, it Already more and more applied to the lens in the research of the disease.Proteomics is understanding of the life activities of the mechanism and the mechanism of disease from proteome level, It is to the research object of whole protein level. two-dimensional electrophoresis is the core technology of proteomics, It is now commonly used only in a piece of gel for separation method of thousands of proteins.Objective:we used two-dimensional gel electrophoresis (2-D DIGE)technique, in combination with DeCyder Differentialln Gel Analysis software to find differential expressed protein spots between opacity degree of lens in diabetic cataract rat and normal rats lens, In order to found differentially expressed proteins. Lay a foundation for the mass spectrometry and Determine the key protein in the disease.Method:we injected STZ and fed high-fat diet for the 20 same-aged SD rats, each group of 10 only,and then take 10 only as control group.we Taken each 10 lens from the group of 6 weeks and 20 weeks of making model,and taken 10 lens in control group as samples. All samples were grinded and the salts were removed out by Clean-up kit, The prepared protein samples were quantified with 2D Quant kit, and then 50μg protein from every sample was minimally labelled with different fluorescent dyes. Cy3, Cy5and Cy2 labelled samples and internal standards were pooled and loaded on a pH gradient strip for isoelectric focusing, equilibrated twice then loaded on polyacrylamide gels for the second electrophoresis. Then quantification and statistic analysis were carried out with DeCyder Differential In Gel Analysis software to find the differential expressed protein.Result:Eventually three groups of samples have 65 differences protein spots, with the aggravation of lens opacity, upgrading expressed in 52 spots and down grading expressed in 32 spots(upgrading expressed or downgrading More than 50% namely 1.5 times,P<0.05 are considered meaningful spots).19 spots have differently expressed in 3 times.Conclusions:Diabetic cataract rats and normal rats existing differences expressed protein of lens, with the aggravation of lens opacity, the quantity of a same protein in each group can express upgrade and downgrade.The differences expressed protein of lens may be relevant with the mechanism of disease diabetic cataract.