CAJ | 학술논문

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)、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ROCK2 mRNA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24 h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(P＜0.05).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RhoA mRNA表达在6h、12h和24 h时均明显增高,血红蛋白组ROCK2 mRNA表达在6h和12 h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.
목적 탐토뇌출혈후혈홍단백시부통과격활RhoA/Rho격매(ROCK)신호통로도치혈뇌병장통투성증가. 방법 70지웅성SD대서안수궤수자표법분위정상대조조(10지)、가수술조(30지)화혈홍단백조(30지),후2조재안조관찰시간점적불동분위6h、12h화24h3개아조(매개아조10지).혈홍단백조채용로내주입혈홍단백법건립대서뇌출혈모형.채용이문사람농도검측평고각조대서혈뇌병장적통투성,채용간습법측정각조대서혈종주위뇌조직적수함량,채용면역조화염색검측RhoA화ROCK2단백재각조대서혈뇌병장적표체분포정황,채용형광정량PCR측정각조대서뇌중RhoA화ROCK2 mRNA적표체정황. 결과 여가수술조화정상대조조상비,혈홍단백조6h、12h화24 h이문사람삼투량화뇌조직함수량균명현승고,차이균유통계학의의(P＜0.05).면역조화염색결과현시:정상대조조혈뇌병장내피세포RhoA화ROCK2미견명현표체,혈홍단백조혈종측혈뇌병장내피세포중RhoA화ROCK2정종황색표체,주요표체우효질세포적세포질;실시형광정량PCR결과현시:여정상대조조화가수술조상비,혈홍단백조RhoA mRNA표체재6h、12h화24 h시균명현증고,혈홍단백조ROCK2 mRNA표체재6h화12 h균명현증고,차이균유통계학의의(P＜0.05). 결론 뇌출혈후혈홍단백가능통과격활RhoA/ROCK신호통로인기혈뇌병장통투성증가,진이삼여뇌수종적발생발전.
Objective To reveal whether increased blood brain barrier (BBB) permeability 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 Hb 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.