华中科技大学学报（医学版）
華中科技大學學報（醫學版）
화중과기대학학보（의학판）
ACTA UNIVERSITATIS MEDICINAE TONGJI
2009年
6期
734-738,743
,共6页
李俐娟%张旻%王雪贞%郝又国%魏佳军%降风%丁志刚%张苏明%卜碧涛
李俐娟%張旻%王雪貞%郝又國%魏佳軍%降風%丁誌剛%張囌明%蔔碧濤
리리연%장민%왕설정%학우국%위가군%강풍%정지강%장소명%복벽도
C型尼曼-皮克病%神经元变性%细胞周期依赖性蛋白激酶-5%轴突球状体%小分子干扰RNA
C型尼曼-皮剋病%神經元變性%細胞週期依賴性蛋白激酶-5%軸突毬狀體%小分子榦擾RNA
C형니만-피극병%신경원변성%세포주기의뢰성단백격매-5%축돌구상체%소분자간우RNA
Niemann-Pick disease type C%neurodegeneration%cyclin dependent kinase 5%axonal spheroid%small in terfering RNA
目的 确定细胞周期依赖性蛋白激酶-5(cyclin dependent kinases-5,cdk5)在C型尼曼-皮克病(Niemann-Pick disease type C,NPC)神经元变性过程中的作用.为NPC的临床治疗提供可能的药物作用靶点.方法 将针对cdk5基因的siRNA序列克隆入rAAV-2载体,对4周龄npc-/-小鼠进行小脑rAAV-cdk5-siRNA-GFP注射(n=8),以空载体病毒(rAAV-GFP)注射组及非手术的同龄npc小鼠为对照(n=8).在干预后4周连续观察小鼠脑内cdk5水平及小鼠神经生化和病理的改变.结果 注射后1周可见针道附近、小脑深部核团及浦肯野细胞出现绿色荧光蛋白表达.与空载体病毒组相比,注射4周后小鼠脑内cdk5的表达水平降低34.17%(P<0.05,n=6);轴突球状体数量减少30.76%(P<0.05,n=6),存活浦肯野细胞数增加300%(P<0.05,n=8);细胞骨架蛋白磷酸化程度降低16.32%(P<0.05,n=6).结论 rAAV2-cdk5-siRNA小脑注射,能显著减轻npc小鼠神经生化和病理改变.cdk5的激活在NPC神经元变性过程中起到了关键性的作用,有可能成为NPC治疗的新靶标.
目的 確定細胞週期依賴性蛋白激酶-5(cyclin dependent kinases-5,cdk5)在C型尼曼-皮剋病(Niemann-Pick disease type C,NPC)神經元變性過程中的作用.為NPC的臨床治療提供可能的藥物作用靶點.方法 將針對cdk5基因的siRNA序列剋隆入rAAV-2載體,對4週齡npc-/-小鼠進行小腦rAAV-cdk5-siRNA-GFP註射(n=8),以空載體病毒(rAAV-GFP)註射組及非手術的同齡npc小鼠為對照(n=8).在榦預後4週連續觀察小鼠腦內cdk5水平及小鼠神經生化和病理的改變.結果 註射後1週可見針道附近、小腦深部覈糰及浦肯野細胞齣現綠色熒光蛋白錶達.與空載體病毒組相比,註射4週後小鼠腦內cdk5的錶達水平降低34.17%(P<0.05,n=6);軸突毬狀體數量減少30.76%(P<0.05,n=6),存活浦肯野細胞數增加300%(P<0.05,n=8);細胞骨架蛋白燐痠化程度降低16.32%(P<0.05,n=6).結論 rAAV2-cdk5-siRNA小腦註射,能顯著減輕npc小鼠神經生化和病理改變.cdk5的激活在NPC神經元變性過程中起到瞭關鍵性的作用,有可能成為NPC治療的新靶標.
목적 학정세포주기의뢰성단백격매-5(cyclin dependent kinases-5,cdk5)재C형니만-피극병(Niemann-Pick disease type C,NPC)신경원변성과정중적작용.위NPC적림상치료제공가능적약물작용파점.방법 장침대cdk5기인적siRNA서렬극륭입rAAV-2재체,대4주령npc-/-소서진행소뇌rAAV-cdk5-siRNA-GFP주사(n=8),이공재체병독(rAAV-GFP)주사조급비수술적동령npc소서위대조(n=8).재간예후4주련속관찰소서뇌내cdk5수평급소서신경생화화병리적개변.결과 주사후1주가견침도부근、소뇌심부핵단급포긍야세포출현록색형광단백표체.여공재체병독조상비,주사4주후소서뇌내cdk5적표체수평강저34.17%(P<0.05,n=6);축돌구상체수량감소30.76%(P<0.05,n=6),존활포긍야세포수증가300%(P<0.05,n=8);세포골가단백린산화정도강저16.32%(P<0.05,n=6).결론 rAAV2-cdk5-siRNA소뇌주사,능현저감경npc소서신경생화화병리개변.cdk5적격활재NPC신경원변성과정중기도료관건성적작용,유가능성위NPC치료적신파표.
Objective To determine the precise role of cyclin dependent kinases-5(cdk5)in Niemann-Pick disease type C (NPC)neurodegeneration,and to offer a new therapeutic target for NPC.Methods Small interfering RNA(siRNA)specific for cdk5 gene was cloned into recombinant adeno-associated virus vector to generate rAAV-cdk5-siRNA-GFP.The rAAV-cdk5- siRNA-GFP and rAAV-GFP were injected into bilateral cerebella of 4-weeks-old npe mice respectively,and non-surgery agematched npc mice were employed as controls(n=8 in each group).The cdk5 levels and neuropathological changes in these mice were observed.Results The expression of GFP could be detected in the path of needle,cerebellar deep nuclei and Purkinje neurons at the first week after injection.Four weeks after injection,the cdk5 level in cerebella was decreased by 34.1 7%(P<0.05,N=6),spheroid number decreased by 30.76%(P<0.05,n=6),number of surviving Purkinje cells increased by 300%(P<GFP can lower the expression of cdk5 gene effectively,reduce the number of axonal spheroids,delay the death of Purkinje neurons and attenuate the hyperphosphorylation of cytoskeletons in npc mice significantly.Cdk5 plays an essential role in the neurodegenerative process and may become a new target for the treatment of NPC.
