cPKCγ参与低氧预适应对小鼠脑缺血皮质内CRMP2水解和磷酸化的调节  

cPKCγ is involved in the regulation of hypoxic preconditioning on CRMP2 hydrolysis and phosphorylation in ischemic cortex of mice

作　　者：刘燕燕[1] 杨璇[1] 韩松[1] 苏吉儿[1] 罗宏[1] 李俊发[1] 

机构地区：[1]首都医科大学神经生物学系北京神经科学研究所,北京100069

出　　处：《基础医学与临床》2012年第1期25-30,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(30871219;31071048;31171147);973计划前期专项(2011CB512109);教育部高等学校博士点科研基金(20091107110001);北京市属高等学校人才强教深化计划创新人才资助(PHR200906116)

摘　　要：目的探讨经典型蛋白激酶Cγ(cPKCγ)在低氧预适应(HPC)调节小鼠脑缺血皮质内脑衰反应蛋白-2(CRMP2)水解和磷酸化中作用。方法利用雄性BALB/c小鼠(18～22 g)HPC和大脑中动脉阻塞(MCAO)模型,借助蛋白印迹、免疫共沉淀和免疫组化等生物化学技术,观察cPKCγ激活对小鼠缺血脑皮质内CRMP2蛋白水解程度(BDP)和磷酸化水平(p-CRMP2)、cPKCγ-CRMP2相互作用和皮质缺血半影区内p-CRMP2阳性细胞数的影响。结果 HPC显著提高缺血脑皮质半影区内p-CRMP2水平,降低BDP产物形成(P<0.05),而侧脑室注射cPKCγ抑制剂Go6983(6nmol/L)可明显解除HPC对半影区内CRMP2蛋白水解和磷酸化的调节作用(P<0.05);免疫共沉淀结果提示,cPKCγ激活程度参与HPC对缺血脑皮质半影区内cPKCγ-CRMP2相互作用的调节;同时,cPKCγ激活与HPC提高脑缺血半影区内p-CRMP2阳性细胞数有关(P<0.05)。结论 cPKCγ参与HPC对小鼠缺血脑皮质内CRMP2水解与磷酸化的调节。Objective To explore the role of cPKCγ in hypoxic preconditioning （HPC） regulating hydrolysis and phosphorylation of collapsin response mediated protein 2 （CRMP2） in the ischemic cortex of mice. Methods Using our established HPC and middle cerebral artery occlusion （MCAO） BALB/c mouse models （male, 18 - 22 g）, We applied Western blot, immunoprecipitation （IP） and immunohistochemisty to determine the effect of cPKC＇y activation on CRMP2 hydrolysis and phosphorylation, cPKCγ-CRMP2 interaction and p-CRMP2 positive cells＇ number in the peri-infarct region of MCAO mice. Results HPC inhibited the decrease of p-CRMP2 level and the increase of BDP in peri-infarct region of ischemic cortex. Pretreatment of cPKCγ inhibitor Go6983 （6nmol/L） could depress the HPC-induced inhibitory effect on CRMP2 dephosphorylation and hydrolysis in peri-infarct area of ischemic cortex; IP results showed that the activity of cPKCγ could affect the interaction between cPKCγ and CRMP2; in addition, the immunohistochemistry results also demonstrated the same effect of cPKCγ on p-CRMP2 positive cells in peri-infarct region of ischemic cortex. Conclusions cPKCγ is involved in the regulation of HPC on CRMP2 phosphorylation and hydrolysis in ischemic cortex of MCAO mice.

关 键 词：缺血/低氧预适应 大脑中动脉阻塞 经典型蛋白激酶C 脑衰反应蛋白-2 

分 类 号：R363[医药卫生—病理学]