神经科学通报（英文版）
神經科學通報（英文版）
신경과학통보（영문판）
NEUROSCIENCE BULLETIN
2008年
5期
288-296
,共9页
葛鹏飞%罗天飞%张纪周%陈大伟%栾永新%付双林
葛鵬飛%囉天飛%張紀週%陳大偉%欒永新%付雙林
갈붕비%라천비%장기주%진대위%란영신%부쌍림
预缺血%蛋白聚集%蛋白伴侣%hsp70
預缺血%蛋白聚集%蛋白伴侶%hsp70
예결혈%단백취집%단백반려%hsp70
ischemic preconditioning%protein aggregation%chaperone%hsp70
目的 研究预缺血对蛋白伴侣hsp70表达和蛋白聚集物形成的影响,探讨其可能的脑保护机制.方法 采用大鼠双侧颈总动脉暂时夹闭法建立全脑缺血模型.大鼠分为3min缺血组,10min缺血组以及预缺血组.苏木素-伊红染色,光镜下随机计数分析预缺血后海马CA1区死亡神经元数量变化.免疫组织化学及激光扫描共聚焦显微镜法观察蛋白伴侣hsp70在CAI区神经元内的分布.差速离心分离细胞浆、细胞核及蛋白聚集物.蛋白印迹法检测不同缺血状态下海马CA1神经元内蛋白聚集物含量的变化,以及胞浆、胞核及蛋白聚集物内蛋白伴侣hsp70含量的变化.结果 组织学检查显示预缺血能够显著减少海马CA1区神经元死亡数量.预缺血诱导海马CA1区神经元内蛋白伴侣hsp70在再灌注后24h表达.预缺血处理后,海马CA1区神经元内蛋白聚集物显著减少.预缺血诱导的蛋白伴侣hsp70与再缺血形成的异常蛋白结合在一起并防止其聚集.结论 预缺血可能通过诱导蛋白伴侣hsp70的表达和抑制再缺血后蛋白聚集物的形成,减少再缺血引起的神经元死亡.
目的 研究預缺血對蛋白伴侶hsp70錶達和蛋白聚集物形成的影響,探討其可能的腦保護機製.方法 採用大鼠雙側頸總動脈暫時夾閉法建立全腦缺血模型.大鼠分為3min缺血組,10min缺血組以及預缺血組.囌木素-伊紅染色,光鏡下隨機計數分析預缺血後海馬CA1區死亡神經元數量變化.免疫組織化學及激光掃描共聚焦顯微鏡法觀察蛋白伴侶hsp70在CAI區神經元內的分佈.差速離心分離細胞漿、細胞覈及蛋白聚集物.蛋白印跡法檢測不同缺血狀態下海馬CA1神經元內蛋白聚集物含量的變化,以及胞漿、胞覈及蛋白聚集物內蛋白伴侶hsp70含量的變化.結果 組織學檢查顯示預缺血能夠顯著減少海馬CA1區神經元死亡數量.預缺血誘導海馬CA1區神經元內蛋白伴侶hsp70在再灌註後24h錶達.預缺血處理後,海馬CA1區神經元內蛋白聚集物顯著減少.預缺血誘導的蛋白伴侶hsp70與再缺血形成的異常蛋白結閤在一起併防止其聚集.結論 預缺血可能通過誘導蛋白伴侶hsp70的錶達和抑製再缺血後蛋白聚集物的形成,減少再缺血引起的神經元死亡.
목적 연구예결혈대단백반려hsp70표체화단백취집물형성적영향,탐토기가능적뇌보호궤제.방법 채용대서쌍측경총동맥잠시협폐법건립전뇌결혈모형.대서분위3min결혈조,10min결혈조이급예결혈조.소목소-이홍염색,광경하수궤계수분석예결혈후해마CA1구사망신경원수량변화.면역조직화학급격광소묘공취초현미경법관찰단백반려hsp70재CAI구신경원내적분포.차속리심분리세포장、세포핵급단백취집물.단백인적법검측불동결혈상태하해마CA1신경원내단백취집물함량적변화,이급포장、포핵급단백취집물내단백반려hsp70함량적변화.결과 조직학검사현시예결혈능구현저감소해마CA1구신경원사망수량.예결혈유도해마CA1구신경원내단백반려hsp70재재관주후24h표체.예결혈처리후,해마CA1구신경원내단백취집물현저감소.예결혈유도적단백반려hsp70여재결혈형성적이상단백결합재일기병방지기취집.결론 예결혈가능통과유도단백반려hsp70적표체화억제재결혈후단백취집물적형성,감소재결혈인기적신경원사망.
Objective To investigate the effect of ischemic preconditioning on chaperone hsp70 expression and protein aggregation in the CA1 neurons of rats,and to further explore its potential neuroprotective mechanism.Methods Two-vesseloccluded transient global ischemia rat model was used.The rats were divided into sublethal 3-min ischemia group,lethal 10rain ischemia group and ischemic preconditioning group.Neuronal death in the CA1 region was observed by hematoxylineosin staining,and number of live neurons was assessed by cell counting under a light microscope.Immunochemistry and laser scanning confocal microscopy were used to observe the distribution of chaperone hsp70 in the CA1 neurons.Differential centrifuge was used to isolate cytosol,nucleus and protein aggregates fractions.Western blot was used to analyze the quantitative alterations of protein aggregates and inducible chaperone hsp70 in cellular fractions and in protein aggregates under different ischemic conditions.Results Histological examination showed that ischemic preconditioning significantly reduced delayed neuronal death in the hippocampus CA1 region(P＜0.01 vs 10-min ischemia group).Sublethal ischemic preconditioning induced chaperone hsp70 expression in the CA1 neurons after 24 h reperfusion following 10-min ischemia.Induced-hsp70 combined with the abnormal proteins produced during the secondary lethal 10-min ischemia and inhibited the formation of cytotoxic protein aggregates(P＜0.01 vs 10-min ischemia group).Conclusion Ischemic preconditioning inducedchaperone hsp70 expression and inhibited protein aggregates formation in the CA1 neurons when suffered secondary lethal ischemia,which may protect neurons from death.
