超声处理对羊瘙痒病脑组织中PrP^(Sc)聚集状态的影响  

Influence of ultrasonic processing on the aggregation of PrP^(Sc) in the brain extracts of the scrapie-infected hamsters

作　　者：周晓勃[1] 高建梅[1] 高晨[1] 张宝云[1] 袁育康[2] 董小平[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所朊病毒室,北京现工作单位100052 [2]西安交通大学医学院免疫教研室

出　　处：《中华实验和临床病毒学杂志》2004年第2期118-121,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家自然科学基金项目 (3 0 0 70 0 3 8) ;国家自然科学基金重点项目 (3 0 13 0 0 70 ) ;国家 863计划项目 (2 0 0 1AA2 15 3 91) ;欧盟项目资助 (QLRT2 0 0 0 0 14 41)

摘　　要：目的　研究超声处理对感染羊瘙痒症仓鼠脑组织中PrPSc聚集状态的影响 ,寻找产生PrPSc低聚体的条件。方法　裂解液制备脑组织提取物 ,用各种超声条件处理不同阶段的脑组织提取物 ;以蛋白酶消化后的Westernblot方法和图象分析系统观测PrPSc蛋白的分布和聚集状态。结果　适当的超声处理 (15s共 30次 )可增加脑组织匀浆上清中PrPSc含量 1 2 9～ 1 5 8倍 ;同样条件下超声处理可明显增加羊瘙痒因子 2 6 3K感染仓鼠脑组织匀浆上清中PrP蛋白总量 ,而对正常对照仓鼠脑组织匀浆上清中PrP蛋白总量影响不大 ;对经常规高速离心获得的PrPSc的超声处理显示约 90 %的PrPSc存在于离心上清液中。结论　对感染动物脑组织进行超声处理可增加PrPSc 的提取量 ,利于实验室检测。适当的超声处理可破碎大的相对分子质量的PrPSc聚集物 ,产生小的相对分子质量的PrPSc产物。Objective To evaluate the influence ultrasonic processing on the aggregation of PrP Sc in the brain extracts prepared from the scrapie-infected hamsters,and to seek for the way to prepare lower molecular PrP Sc polymer. Methods The extracts of infected brains were prepared with a lysis solution,and treated with ultrasonics at various conditions during the different phases. The distribution and aggregation state of PrP Sc were analyzed by proteinase K treated Western blot,and afterwards,quantitatively calculated with a commercially supplied software Image Totaltech. Results The amount of PrP Sc in the supernatant of brain homogenates was 1.29-to 1.58-fold increased with appropriate sonication (15 s for 30 times). At the same conditions of ultrasound,the PrP amount in the supernatant prepared from the scrapie-infected hamster′s brain was significantly increased,whereas that prepared from healthy animal used as normal control showed little change. Comparative analyses of PrP Sc pellets prepared by high-speed centrifugation revealed that about 90% PrP Sc released into supernatant after ultrasound processing. Conclusion Appropriate sonication of homogenate of scrapie-infected brain increases the extracted amount of PrP Sc ,being favorable to laboratory diagnosis. Larger molecular PrP Sc aggregates can be crashed by ultrasonic processing,engendering lower molecular PrP Sc polymers.

关 键 词：超声处理 羊 瘙痒病 脑组织 PrP^Sc蛋白 聚集状态 可传播性海绵样变脑病 

分 类 号：S854.5[农业科学—临床兽医学]