cPKCgamma参与低氧预适应对小鼠脑缺血皮层内CRMP2水解和磷酸化的调节

基础医学与临床 ›› 2012, Vol. 32 ›› Issue (1): 25-30.

cPKCgamma参与低氧预适应对小鼠脑缺血皮层内CRMP2水解和磷酸化的调节

刘燕燕¹,杨璇¹,韩松¹,苏吉儿¹,罗宏¹,李俊发²

1. 首都医科大学神经生物学系
2. 首都医科大学

收稿日期:2011-10-09 修回日期:2011-11-24 出版日期:2012-01-05 发布日期:2011-12-28
通讯作者: 李俊发 E-mail:junfali@ccmu.edu.cn
基金资助:
国家自然科学基金;国家自然科学基金;国家自然科学基金;973计划前期专项;教育部高等学校博士点科研基金;北京市属高等学校人才强教深化计划创新人才资助

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

Received:2011-10-09 Revised:2011-11-24 Online:2012-01-05 Published:2011-12-28
Contact: Jun-fa LI E-mail:junfali@ccmu.edu.cn
Supported by:
The National Natural Science Foundation of China;The National Natural Science Foundation of China;The National Natural Science Foundation of China

摘要/Abstract

摘要： 摘要：目的探讨经典型蛋白激酶Cgamma( cPKCgamma)在低氧预适应(HPC)调节小鼠脑缺血皮层内脑衰反应蛋白-2(CRMP2)水解和磷酸化中作用。方法利用雄性BALB/c小鼠(18-22g) 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cPKCgamma激活对小鼠缺血脑皮层内CRMP2蛋白水解程度（BDP）和磷酸化水平（p-CRMP2）、cPKCgamma-CRMP2相互作用和皮层缺血半影区内p-CRMP2阳性细胞数的影响。结果　我们发现，HPC显著提高缺血脑皮层半影区内p-CRMP2水平，降低BDP产物形成，而侧脑室注射cPKCgamma抑制剂Go6983(6nM)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；免疫共沉淀结果提示，cPKCgamma激活程度参与HPC对缺血脑皮层半影区内cPKCgamma-CRMP2相互作用的调节；同时，免疫组化进一步证实，cPKCgamma激活与HPC提高脑缺血半影区内p-CRMP2阳性细胞数有关。结论 cPKCgamma参与HPC对小鼠缺血脑皮层内CRMP2水解与磷酸化的调节。

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

Abstract: Abstract: Objective To explore the role of cPKCgamma 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,18-22g), We applied Western blot, immunoprecipitation (IP) and immunohistochemisty to determine the effect of cPKCgamma activation on CRMP2 hydrolysis and phosphorylation, cPKCgamma-CRMP2 interaction and p-CRMP2 positive cells’ number in the peri-infarct region of MCAO mice. Results We found that HPC could inhibit both the decrease of p-CRMP2 level and increase of BDP in peri-infarct region of ischemic cortex. Pretreatment of cPKCgmma inhibitor Go6983 (6nM) 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 cPKCgamma could affect the interaction between cPKCgamma and CRMP2; in addition, the immunohistochemistry results also demonstrated the same effect of cPKCgamma on p-CRMP2 positive cells in peri-infarct region of ischemic cortex. Conclusion cPKCgamma was involved in the regulation of HPC on CRMP2 phosphorylation and hydrolysis in ischemic cortex of MCAO mice.

Key words: HPC, middle cerebral artery occlusion, cPKC?, CRMP2

中图分类号:

R339.5

刘燕燕杨璇韩松苏吉儿罗宏李俊发. cPKCgamma参与低氧预适应对小鼠脑缺血皮层内CRMP2水解和磷酸化的调节[J]. 基础医学与临床, 2012, 32(1): 25-30.

[1]	杜建丽梁婧李沫音张旭赵丽罗艳琳李俊发. MSK-1和CREB参与低氧预适应降低小鼠缺血性脑损伤[J]. 基础医学与临床, 2010, 30(11): 1143-1148.
[2]	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.