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目的:比较由葡萄糖和果糖诱发不同的人胚胎γ-晶体蛋白的非酶糖化作用。方法:用Sephadex凝胶层析方法从人类胚胎晶体中分离出γ_1和γ_3-晶体蛋白,将这些γ-晶体蛋白与葡萄糖或果糖37℃下培养20天,γ-晶体蛋白产生非酶糖化作用,混浊和块状沉淀出现,还观察到SDS—PAGE图谱的变化和蓝色荧光产物。结果:虽然两种γ-晶体蛋白出现相类似的变化。但在早期阶段(3天)γ_1-晶体蛋白更容易产生聚合和不溶解。SDS—PAGE显示有聚合物产生,这些都是通过二硫键和非二硫键交联以及肽键的降解而形成。γ_3-晶体蛋白溶液糖化作用后,蓝色荧光增加。果糖与γ-晶体蛋白的糖化作用比葡萄糖更明显(如:聚合、降解、产生蓝色荧光)。结论:人类胚胎γ_1和γ_3-晶体蛋白对非酶糖化作用是敏感的,程度亦有不同(γ_1>γ_3),果糖的作用比葡萄糖强。眼科学报 1995;11:197—201
OBJECTIVE: To compare the non-enzymatic glycation of different human embryonic γ-crystallin induced by glucose and fructose. Methods: γ_1 and γ_3-crystal proteins were isolated from human embryo crystals by Sephadex gel chromatography. The γ-crystal proteins were non-enzymatic glycated by incubation with glucose or fructose for 20 days at 37 ° C. Turbidity and massive deposits appeared, changes in the SDS-PAGE profile and blue fluorescent products were also observed. Results: Similar changes occurred in both γ-crystallins. In the early stages (3 days), however, γ-1-crystallin is more prone to polymerization and insolubilization. SDS-PAGE showed polymer production, all of which were formed by cross-linking of disulfide and non-disulfide bonds and degradation of peptide bonds. After the γ_3-crystal protein solution saccharification, the blue fluorescence increased. The saccharification of fructose and γ-crystallin is more pronounced than glucose (eg, polymerization, degradation, producing blue fluorescence). CONCLUSION: Human embryonic γ_1 and γ_3-crystal proteins are sensitive to non-enzymatic glycation with varying degrees (γ_1> γ_3), with the effect of fructose being stronger than that of glucose. Journal of Ophthalmology 1995; 11: 197-201