| 脑衰蛋白反应调节蛋白-2参与低氧预适应减轻小鼠脑缺血损伤 |
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| 引用本文： | 张彩艳,封素娟,刘旭,卜祥宁,张楠,郑雅欣,袁晓文,李晓光,李俊发. 脑衰蛋白反应调节蛋白-2参与低氧预适应减轻小鼠脑缺血损伤[J]. 基础医学与临床, 2009, 29(11): 1133-1138 |
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| 作者姓名： | 张彩艳 封素娟 刘旭 卜祥宁 张楠 郑雅欣 袁晓文 李晓光 李俊发 |
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| 作者单位： | 首都医科大学,神经生物学系,北京神经科学研究所,北京,100069;首都医科大学,神经生物学系,北京神经科学研究所,北京,100069;首都医科大学,神经生物学系,北京神经科学研究所,北京,100069;首都医科大学,神经生物学系,北京神经科学研究所,北京,100069;首都医科大学,神经生物学系,北京神经科学研究所,北京,100069;首都医科大学,神经生物学系,北京神经科学研究所,北京,100069;首都医科大学,神经生物学系,北京神经科学研究所,北京,100069;首都医科大学,神经生物学系,北京神经科学研究所,北京,100069;首都医科大学,神经生物学系,北京神经科学研究所,北京,100069 |
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| 基金项目： | 国家自然科学基金，国家重点基础研究发展规划(973计划)，北京市自然科学基金，北京市教育委员会科技计划重点项目，北京市属高等学校人才强教计划项目，北京市新世纪百千万人才工程培养经费资助 |
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| 摘 要： | 目的 研究参与低氧预适应(HPC)的经典型蛋白激酶C II(cPKC II)相互作用的脑衰蛋白反应调节蛋白-2(CRMP-2) 对缺血脑是否有保护作用。方法 成年雄性BALB/c小鼠随机分为常氧(Nor)、HPC、常氧假手术(Nor+sham)、HPC假手术(HPC+sham)、常氧缺血(Nor+I)和HPC缺血(HPC+I)等6组(每组n=6)。应用小鼠整体HPC和脑中动脉梗死(MCAO)致脑局部缺血模型，结合免疫沉淀、双向凝胶电泳和质谱等技术，分离和鉴定与cPKC II相互作用蛋白；利用聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白印迹(Western blot)技术，分析CRMP-2磷酸化和蛋白降解水平在脑HPC和缺血中的变化。结果 与Nor组比，HPC鼠脑皮层组织内有10种与cPKC? II相互作用蛋白的表达发生了明显变化，其中CRMP-2在膜相关蛋白组分中的表达量升高，而在胞质蛋白组分中的表达量降低。在脑缺血模型中，与Nor+Sham组相比，Nor+I组小鼠脑皮层缺血核心区(Ic)CRMP-2磷酸化水平明显降低(p<0.05, n=6)；与Nor+I组相比，HPC+I组小鼠脑皮层Ic区内CRMP-2磷酸化水平明显增高(p<0.05, n=6)。脑缺血可导致CRMP-2发生水解并伴随着大量55-ku水解片段(BDP)的出现，但与Nor+I组相比，HPC+I组小鼠脑皮层缺血半影区(P)内CRMP-2水解片段减少，水解率明显降低(p<0.05, n=6)。结论 CRMP-2参与了 HPC缓解小鼠脑缺血Ic区内CRMP-2磷酸化水平的降低和减少P区内CRMP-2水解片段从而减轻缺血脑组织的损伤。
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| 关 键 词： | 蛋白激酶CβⅡ 脑衰蛋白反应调节蛋白-2(CRMP-2) 蛋白质组学 低氧预适应 大脑中动脉阻塞 |
| 收稿时间： | 2009-07-27 |
| 修稿时间： | 2009-08-31 |
CRMP-2 is involved in hypoxic preconditioning-induced neuroprotection against cerebral ischemic injuries of mice |
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| ZHANG Cai-yan,FENG Su-juan,Liu Xu,BU Xiang-ning,ZHANG Nan,ZHENG Ya-xin,YUAN Xiao-wen,LI Xiao-guang,LI Jun-fa. CRMP-2 is involved in hypoxic preconditioning-induced neuroprotection against cerebral ischemic injuries of mice[J]. Basic Medical Sciences and Clinics, 2009, 29(11): 1133-1138 |
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| Authors: | ZHANG Cai-yan FENG Su-juan Liu Xu BU Xiang-ning ZHANG Nan ZHENG Ya-xin YUAN Xiao-wen LI Xiao-guang LI Jun-fa |
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| Abstract: | Objective To investigate whether conventional protein kinase C (cPKC ) βⅡ-interacting collapsin response mediator protein-2 (CRMP-2) provides neuroprotection against cerebral ischemic (I) injuries. Methods Male BALB/c mice were randomly divided into normoxic control (Nor) , HPC, Nor + Sham, HPC + Sham, Nor + I and HPC + I groups (n = 6 per group). Using our HPC and MCAO mouse models, we applied immunoprecipita-tion, two-dimensional electrophoresis and mass spectrometry to characterize cPKCβⅡ-interacting proteins and combined with SDS-PAGE and Western blot to quantitatively analyze CRMP-2 phosphorylation and degradation levels in the brain of mice after HPC and MCAO. Results The expression level of 10 cPKCβⅡ-interacting proteins changed obviously in cerebral cortex of HPC mice when compared with Nor group. One of these proteins, CRMP-2 protein level increased in particulate fraction and decreased in cytosolic fraction of cerebral cortex of HPC mice. CRMP-2 phosphorylation level in ischemic core (Ic) of cerebral cortex decreased significantly ( P < 0. 05 , n = 6) as compared with that of Nor + sham group, but CRMP-2 phosphorylation level in HPC +I group increased significantly as compared with that of Nor +I group ( P < 0. 05, n = 6). In ischemic cortex, CRMP-2 degradation (proteolysis) was observed as the appearance of 55 ku breakdown products (BDP). However, the CRMP-2 degradation level, BDPs products decreased significantly in penumbra ( P) of ischemic cortex from HPC +I group when we compared with that of Nor +I group (P < 0. 05, n = 6 ). Conclusion CRMP-2 is involved in attenuating the decrease of CRMP-2 phosphorylation in ischemic core and in inhibiting its degradation in penumbra of cerebral cortex of mice thereby to lessen the ischemic injuries. |
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| Keywords: | cPKCβⅡ collapsin response mediator protein-2 ( CRMP-2) proteomics hypoxic preconditioning ( HPC) middle cerebral artery occlusion (MCAO) |
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