Cdk5及p35在星形胶质细胞正常及缺血再灌注后的表达研究
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摘要
目的:研究细胞周期蛋白依赖性激酶(Cdk)5和p35在星形胶质细胞中的表达及缺血再灌注后的其表达变化趋势。方法:离体培养SD大鼠星形胶质细胞,通过免疫荧光染色观察Cdk5和p35在星形胶质细胞中的表达分布。对培养的星形胶质细胞进行糖氧剥夺/恢复(OGD/R)处理,Western blot检测Cdk5和p35在星形胶质细胞OGD/R后不同时间点的表达变化。结果:Cdk5和p35均在星形胶质细胞中存在表达,以胞浆为主,胞核表达较少;两者表达分布一致。OGD/R后Cdk5和p35的表达呈早期显著上升,后期下降的趋势。结论:Cdk5和p35在星形胶质细胞中均存在表达,以胞浆为主,且两者表达分布基本一致。OGD/R后星形胶质细胞中的Cdk5和p35的表达呈动态变化。
Objective: To investigate the expression of Cdk5 and p35 in primary cultured astrocytes and the changes in expression induced by ischemia/reperfusion. Methods: Astrocytes were cultured from SD neonate rats. The expression and distribution of Cdk5 and p35 in cultured astrocytes were analyzed by immunofluorescence staining. Primary cultured astrocytes were subjected to oxygen-glucose deprivation/reperfusion(OGD/R), and the changes in Cdk5 and p35 expression after OGD/R were observed by Western blot.Results: Cdk5 and p35 were expressed in astrocytes, mainly in the cytoplasm and less in the nucleus, and both displayed similar cellular distribution. The expression of Cdk5 and p35 in astrocytes was increased during early phases and decreased during late phases after OGD/R. Conclusion: Both Cdk5 and p35 were expressed in astrocytes, and both showed similar, mainly cytoplasmic distribution. Cdk5 and p35 expression in astrocytes exhibited dynamic changes after OGD/R.
Objective: To investigate the expression of Cdk5 and p35 in primary cultured astrocytes and the changes in expression induced by ischemia/reperfusion. Methods: Astrocytes were cultured from SD neonate rats. The expression and distribution of Cdk5 and p35 in cultured astrocytes were analyzed by immunofluorescence staining. Primary cultured astrocytes were subjected to oxygen-glucose deprivation/reperfusion(OGD/R), and the changes in Cdk5 and p35 expression after OGD/R were observed by Western blot.Results: Cdk5 and p35 were expressed in astrocytes, mainly in the cytoplasm and less in the nucleus, and both displayed similar cellular distribution. The expression of Cdk5 and p35 in astrocytes was increased during early phases and decreased during late phases after OGD/R. Conclusion: Both Cdk5 and p35 were expressed in astrocytes, and both showed similar, mainly cytoplasmic distribution. Cdk5 and p35 expression in astrocytes exhibited dynamic changes after OGD/R.
引文
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[14]Ko J,Humbert S,Bronson RT,et al.p35 and p39 are essential for cyclin-dependent kinase 5 function during neurodevelopment[J].J Neurosci,2001,21:6758-6771.
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[16]Meyer DA,Torres-Altoro MI,Tan Z,et al.Ischemic stroke injury is mediated by aberrant Cdk5[J].J Neurosci,2014,34:8259-8267.
[17]Ji YB,Zhuang PP,Ji Z,et al.TFP5 peptide,derived from CDK5-activating cofactor p35,provides neuroprotection in early-stage of adult ischemic stroke[J].Sci Rep,2017,7:40013.
[18]Plattner F,Hernández A,Kistler TM,et al.Memory enhancement by targeting Cdk5 regulation of NR2B[J].Neuron,2014,81:1070-1083.
[19]Lai TW,Zhang S,Wang YT.Excitotoxicity and stroke:identifying novel targets for neuroprotection[J].Prog Neurobiol,2014,115:157-188.
[20]Becerra-Calixto A,Cardona-Gómez GP.The Role of Astrocytes in Neuroprotection after Brain Stroke:Potential in Cell Therapy[J].Front Mol Neurosci,2017,10:88-92.
[21]Choudhury GR,Ding S.Reactive astrocytes and therapeutic potential in focal ischemic stroke[J].Neurobiol Dis,2016,85:234-244.
[2]Wang W,Redecker C,Yu ZY,et al.Rat focal cerebral ischemia induced astrocyte proliferation and delayed neuronal death are attenuated by cyclin-dependent kinase inhibition[J].J Clin Neurosci,2008,15:278-285.
[3]Zhu Z,Zhang Q,Yu Z,et al.Inhibiting cell cycle progression reduces reactive astrogliosis initiated by scratch injury in vitro and by cerebral ischemia in vivo[J].Glia,2007,55:546-558.
[4]Malumbres M.Cyclin-dependent kinases[J].Genome Biol,2014,15:122-126.
[5]Mita N,He X,Sasamoto K,et al.Cyclin-Dependent Kinase 5Regulates Dendritic Spine Formation and Maintenance of Cortical Neuron in the Mouse Brain[J].Cereb Cortex,2016,26:967-976.
[6]Tanaka T,Veeranna,Ohshima T,et al.Neuronal cyclin-dependent kinase 5 activity is critical for survival[J].J Neurosci,2001,21:550-558.
[7]He Y,Li HL,Xie WY,et al.The presence of active Cdk5 associated with p35 in astrocytes and its important role in process elongation of scratched astrocyte[J].Glia,2007,55:573-583.
[8]Liu R,Tian B,Gearing M,et al.Cdk5-mediated regulation of the PIKE-A-Akt pathway and glioblastoma cell invasion[J].Proc Natl Acad Sci U S A,2008,105:7570-7575.
[9]Shah K,Lahiri DK.Cdk5 activity in the brain-multiple paths of regulation[J].J Cell Sci,2014,127:2391-2400.
[10]Ino H,Chiba T.Intracellular localization of cyclin-dependent kinase 5(CDK5)in mouse neuron:CDK5 is located in both nucleus and cytoplasm[J].Brain Res,1996,732:179-185.
[11]Liu SL,Wang C,Jiang T,et al.The Role of Cdk5 in Alzheimer's Disease[J].Mol Neurobiol,2016,53:4328-4342.
[12]Luo F,Zhang J,Burke K,et al.The Activators of Cyclin-Dependent Kinase 5 p35 and p39 Are Essential for Oligodendrocyte Maturation,Process Formation,and Myelination[J].J Neurosci,2016,36:3024-3037.
[13]Fang-Hu,Zhang HH,Yang BX,et al.Cdk5 contributes to inflammation-induced thermal hyperalgesia mediated by the p38 MAPK pathway in microglia[J].Brain Res,2015,1619:166-1675.
[14]Ko J,Humbert S,Bronson RT,et al.p35 and p39 are essential for cyclin-dependent kinase 5 function during neurodevelopment[J].J Neurosci,2001,21:6758-6771.
[15]Shupp A,Casimiro MC,Pestell RG.Biological functions of CDK5and potential CDK5 targeted clinical treatments[J].Oncotarget,2017,8:17373-17382.
[16]Meyer DA,Torres-Altoro MI,Tan Z,et al.Ischemic stroke injury is mediated by aberrant Cdk5[J].J Neurosci,2014,34:8259-8267.
[17]Ji YB,Zhuang PP,Ji Z,et al.TFP5 peptide,derived from CDK5-activating cofactor p35,provides neuroprotection in early-stage of adult ischemic stroke[J].Sci Rep,2017,7:40013.
[18]Plattner F,Hernández A,Kistler TM,et al.Memory enhancement by targeting Cdk5 regulation of NR2B[J].Neuron,2014,81:1070-1083.
[19]Lai TW,Zhang S,Wang YT.Excitotoxicity and stroke:identifying novel targets for neuroprotection[J].Prog Neurobiol,2014,115:157-188.
[20]Becerra-Calixto A,Cardona-Gómez GP.The Role of Astrocytes in Neuroprotection after Brain Stroke:Potential in Cell Therapy[J].Front Mol Neurosci,2017,10:88-92.
[21]Choudhury GR,Ding S.Reactive astrocytes and therapeutic potential in focal ischemic stroke[J].Neurobiol Dis,2016,85:234-244.