αB-晶状体蛋白在半乳糖性白内障大鼠眼晶状体中的表达

Expression of alpha B-crystallin in galactosemic cataractous rat lenses

目的 检测轻、中、重度混浊的半乳糖性白内障大鼠晶状体中可溶性αB 晶状体蛋白和不溶性αB 晶状体蛋白的表达 ,探讨αB 晶状体蛋白与白内障发生的关系。方法 采用 50 %半乳糖注射液腹腔注射诱发半乳糖性白内障作为实验组 ,依晶状体混浊程度分为轻、中、重度三组 ,对照组则同时腹腔注射生理盐水 ,与实验组大鼠同期分为三组。大鼠处死后 ,取晶状体作匀浆 ,其上清 (可溶性晶状体蛋白质 )和沉淀 (不溶性晶状体蛋白质 )分别用免疫印迹法 (Westernblotanalysis)检测αB 晶状体蛋白的表达。结果 实验组可溶性αB 晶状体蛋白含量较对照组降低 ,有显著性差异 (P <0 0 5) ;实验组含量随晶状体混浊程度加重而逐渐降低 ,其差别有显著性意义 (P <0 0 5)。不溶性αB 晶状体蛋白含量较对照组升高 ,有显著性差异 (P <0 0 5) ;实验组含量随晶状体混浊程度加重而逐渐升高 ,其差别有显著性意义 (P <0 0 1)。结论 αB 晶状体蛋白的分子伴侣功能参与白内障的发病 。

ObjectiveTo examine the expression of soluble α B -crystallin and insoluble α B-crystallin in galactosemic cataractous rat lens ,and to probe the correlation between α B-crystallin and the pathogenesis of c ataract.MethodsExperimental group of galactosemic cataract rats was induced by 50% galactogen daily intraperitoneal injection.The physiological saline was i ntraperitoneally injected in corresponding control group.All lens were examined using a slit lamp everyday.The experimental group was further divided into A,B,C group according to their different degree of cataract and the experimental cont rol group into A,B,C group as well.After the rats were sacrificed,the lenses we re isolated carefully and homogenized in a cold buffer containing 0 1 mol/L so dium phosphate (pH7 4) and 0 1 mol/L NaCl.Lens homogenate was divided into so luble and insoluble fractions by centrifugating at 13 000 g at 4℃ for 20 m inutes.Both the supernatant and the pellet were obtained for the detection of α B-crystallin with Western blot analysis.ResultsCompared with the correspon ding control group,the soluble α B-crystallin level of each experimental grou p was markedly decreased ( P <0 05),but the insoluble α B-crystallin leve l was markedly increased ( P <0 05).In the three experimental groups,the lev els of soluble α B-crystallin were decreased, and the levels of insoluble α B -crystallin were increased as the lens opacity developed ( P <0 01).Concl usionThe molecular chaperone function of α B-crystallin is involved in the p athogenesis of cataract,which provides a new way to the further study of pathog enesis of cataract.