脑内源性cPKC gamma水平影响缺血性卒中小鼠脑梗死体积和神经功能损伤  

Endogenous cPKC gamma protein levels affect cerebral infarction volume and neurological outcome of mice with ischemic stroke

作　　者：李洁霏[1] 章欣欣[1] 钟洁[1] 杨璇[1] 李筠[1] 韩松[1] 李俊发[1] 

机构地区：[1]首都医科大学神经生物学系北京脑重大疾病研究院,北京100069

出　　处：《基础医学与临床》2013年第11期1452-1459,共8页Basic and Clinical Medicine

基　　金：国家自然科学基金(31071048;31171147);科技部973计划前期研究专项(2011CB512109)

摘　　要：目的探讨脑内源性经典型蛋白激酶C（cPKC）γ表达水平对脑中动脉阻塞（MCAO）／再灌注（R）致缺血性脑卒中小鼠脑梗死体积和神经功能损伤程度的影响。方法利用cPKCγ基因野生型（WT）、杂合型（HET）和敲除型（KO）小鼠，建立小鼠1hMCAO／R24h和7d缺血性脑卒中模型，借助Westernblot蛋白印迹、2，3，5-氯化三苯基四唑（TTC）染色、神经行为学测试等技术方法，检测脑内源性cPKCγ蛋白表达水平、脑梗死体积和神经功能损伤情况。结果MCAO／R1h／24h和7d可使WT、HET和KO小鼠脑出现明显的梗死灶和神经功能损伤；具有双拷贝基因的wT小鼠脑内cPKCγ／蛋白表达量存在个体差异的同时，只有单拷贝基因的HET小鼠脑内cPKCγ蛋白表达水平约为wT型小鼠的30％～100％，而非简单的50％；脑内源性cPKCγ／蛋白表达量与卒中脑梗死体积大小呈明显的负相关性，且神经功能损伤程度随着脑内cPKCγ／蛋白表达水平的增高而明显减轻。结论脑内源性cPKCγ／蛋白表达水平可影响缺血性卒中小鼠脑梗死体积和神经功能损伤程度。Objective To explore the effect of endogenous conventional protein kinase C （cPKC） γ/ protein expression level on cerebral infarction volume and neurological outcome of mice following ischemic stroke. Methods By using cPKC＇y gene wild type （WT） , heterogeneous （HET） and knockout （KO） mice, the 1 h middle cerebral artery occlusion （ MCAO）/reperfusion （R） 24 h and 7 d-induced ischemic stroke model was de- veloped, we applied the techniques of Western blot, 2,3,5-triphenyhetrazolium chloride （ TTC ） staining and neurological behavior tests to determine endogenous cPKCγ/ protein expression levels, cerebral infarction volume and neurological outcome, respectively. Results The treatments of 1 h MCAO/R 24 h and 7 d induced cerebral infarction and neurological dysfunction of WT, HET and KO mice. There was individual differences in endoge- nous cPKCγ/protein expression levels among WT mice with two copies of cPKCγgene, while the endogenous cPKCγ/protein expression levels of HET mice with single copy of cPKC＇y gene could reach about 30% - 100% ,not simply 50% of the WT mice. The cerebral endogenous cPKCγ/protein expression level was correlated nega- tively with the infarction size, the neurological outcome was improved with the increase of endogenous cPKCγ/pro- tein expression levels in 1 h MCAO/R 24 h and 7 d treated mice. Conclusions Cerebral endogenous cPKCγ protein expression level affects the brain infarction size and the neurological outcome of mice with ischemic stroke.

关 键 词：缺血性脑卒中 蛋白激酶C（PKC） 脑梗死体积 神经功能损伤程度 基因敲除小鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学]