RhoA／Rho激酶信号通路在脑出血后血红蛋白影响血脑屏障通透性中的作用  

Effect of RhoA/Rho associated coiled coil forming protein kinase signal pathway on blood brain barrier at early stage of cerebral hemorrhage after intracerebral hemoglobin injection

作　　者：付正浩[1] 陈祎招[1] 杨硕[1] 邓心情[1] 张润[1] 丁锐[1] 冯亮[1] 张世忠[1] 柯以铨[1] 

机构地区：[1]南方医科大学珠江医院神经外科广东神经外科研究所广东省脑功能修复与再生蓖点实验室,广州510282

出　　处：《中华神经医学杂志》2013年第12期1244-1248,共5页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（30500526、81271314）;广东省A然科学基金（5300468）;广东省教育部产学研结合项目（20128091100154）;国家临床重点建设专科资助项目

摘　　要：目的探讨脑出血后血红蛋白是否通过激活RhoA／Rho激酶（ROCK）信号通路导致血脑屏障通透性增加。方法70只雄性SD大鼠按随机数字表法分为正常对照组（10只1、假手术组（30只）和血红蛋白组（30只），后2组再按照观察时间点的不同分为6h、12h和24h3个亚组（每个亚组10只）。血红蛋白组采用颅内注入血红蛋白法建立大鼠脑出血模型。采用伊文思蓝浓度检测评估各组大鼠血脑屏障的通透性，采用干湿法测定各组大鼠血肿周围脑组织的水含量．采用免疫组化染色检测RhoA和ROCK2蛋白在各组大鼠血脑屏障的表达分布情况，采用荧光定量PCR测定各组大鼠脑中RhoA和ROCK2mRNA的表达情况。结果与假手术组和正常对照组相比．血红蛋白组6h、12h和24h伊文思蓝渗透量和脑组织含水量均明显升高，差异均有统计学意义（RO．05）。免疫组化染色结果显示：正常对照组血脑屏障内皮细胞RhoA和ROCK2未见明显表达，血红蛋白组血肿侧血脑屏障内皮细胞中RhoA和ROCK2呈棕黄色表达，主要表达于胶质细胞的细胞质：实时荧光定量PCR结果显示：与正常对照组和假手术组相比．血红蛋白组RhoAmRNA表达在6h、12h和24h时均明显增高，血红蛋白组ROCK2mRNA表达在6h和12h均明显增高．差异均有统计学意义（P〈0．05）。结论脑出血后血红蛋白可能通过激活RhoA／ROCK信号通路引起血脑屏障通透性增加，进而参与脑水肿的发生发展。Objective To reveal whether increased blood brain barrier （BBB） perrneabihty induced by hemoglobin is attributed to RhoA/Rho associated coiled coil forming protein kinase （ROCK） signal pathway activation. Methods Seventy male SD rats were randomized into normal control group （n=10）, sham-operated group （n=30） and intracerebral hemoglobin injection group （Hb group, n=30）. Rats in sham-operated group and Hb group were each divided into three subgroups （6, 12 and 24 h after hemoglobin injection, n=10）. Rat models of cerebral hemorrhage in the Hb group were induced by intracerebral hemoglobin injection; BBB permeability was measured by Evans blue dye; dry-wet method was used to measure the brain tissue water content for all groups; the distributions of RhoA and ROCK2 were detected by immunohistochemistry （DAB stainning）; the mRNA expressions of RhoA and ROCK2 were evaluated by real time fluorescent quantitative PCR. Results As compared with those in the sham-operated group and normal control group, Evans blue permeation and water content of perihematoma significantly increased in the Hb group at all time points （P〈0.05）. Immunohistochemistry indicated no obvious expressions of RhoA and ROCK2 in the normal control group, while RhoA and ROCK2 showed claybank in the brain tissue of lib group, which mainly expressed in the cytoplasm. As compared with those in the normal control group and sham-operated group, the mRNA level of RhoA significantly elevated in the Hb group at all time points （P〈0.05） and that of ROCK2 significantly elevated in the Hb group at 6 and 12 h time points （P〈0.05）. Conclusion Hemoglobin induces RhoA/ROCK signal pathway activation after intracerebral hemorrhage, which may attribute to BBB permeability increase and subsequent cerebral edema formation.

关 键 词：脑出血 血红蛋白 RHOA Rho激酶信号通路 血脑屏障 

分 类 号：R743.34[医药卫生—神经病学与精神病学]