p38 MAPK通路对大鼠脑积水及水通道蛋白4表达的影响  

Effect of p38 mitogen-activated protein kinase pathway on hydrocephalus and aquaporin 4 expression in rats

作　　者：李张昱 高鑫[1] 毛建耀 陈四方[1] 谭国伟[1] 王占祥[1] Li Zhangyu;Gao Xin;Mao Jianyao;Chen Sifang;Tan Guowei;Wang Zhanxiang(Department of Neurosurgery,First Affiliated Hospital of Xiamen University,Xiamen 361003,China)

机构地区：[1]厦门大学附属第一医院神经外科,厦门361003

出　　处：《中华神经医学杂志》2022年第4期333-340,共8页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金面上项目(81271332);厦门市重要重大疾病联合攻关项目(3502Z20159021)。

摘　　要：目的探讨脑积水后p38丝裂原活化蛋白激酶(p38 MAPK)对水通道蛋白4(AQP4)的调控作用及其在脑积水防治中的意义。方法50只SD大鼠按照随机数字表法分为3组:假手术组10只,脑积水组20只,脑积水+抑制剂组(后文简称为SB组)20只。SB组及脑积水组大鼠均采用侧脑室注射高岭土方法构建脑积水模型,假手术组大鼠侧脑室注射等量生理盐水;SB组大鼠进一步于造模后第8天予腹腔注射p38 MAPK特异性抑制剂SB203580(10 mg/kg),连续注射7 d;脑积水组同期注射等量DMSO稀释液,假手术组大鼠不做处理。通过MRI观察3组大鼠脑积水严重程度;采用酶联免疫吸附实验(ELISA)检测3组大鼠脑脊液中炎性因子肿瘤坏死因子-α(TNF-α)水平;采用实时荧光定量PCR(qRT-PCR)检测3组大鼠脑组织AQP4 mRNA和TNF-αmRNA表达水平;采用Western blotting实验及免疫组织化学染色检测3组大鼠室周区域脑组织AQP4及磷酸化p38 MAPK表达水平。结果造模后脑积水组和SB组大鼠均出现脑积水。与假手术组相比,脑积水组大鼠、SB组大鼠侧脑室体积增大,室周水肿加重,EVAN’s指数升高,脑组织含水量增多,差异均有统计学意义(P<0.05)。造模后2周假手术组、脑积水组、SB组大鼠脑脊液中TNF-α表达量分别为(20.49±0.96)、(42.04±3.17)、(28.00±3.71)pg/mL,差异有统计学意义(F=186.000,P<0.001);其中SB组大鼠TNF-α表达量明显高于假手术组,低于脑积水组,差异均有统计学意义(P<0.05)。造模后2周3组大鼠脑组织TNF-αmRNA、AQP4 mRNA表达水平差异均有统计学意义(P<0.05);其中脑积水组TNF-αmRNA、AQP4 mRNA表达水平均较假手术组和SB组明显升高,差异均有统计学意义(P<0.05)。相关性分析结果显示脑积水组AQP4 mRNA与TNF-αmRNA表达水平呈正线性相关关系(r=0.511,P=0.026)。造模后2周3组大鼠室周脑组织AQP4蛋白及磷酸化p38 MAPK蛋白表达水平差异均有统计学意义(P<0.05),其中脑积水组AQP4蛋白及磷�Objective To observe the regulatory effect of p38 mitogen-activated protein kinase(p38 MAPK)on aquaporin 4(AQP4)in rats after hydrocephalus,and to explore its significance in hydrocephalus prevention.Methods Fifty SD rats were randomly divided into sham-operated group(n=10),hydrocephalus group(n=20),and hydrocephalus+inhibitor(SB203580)group(SB group,n=20).The rat models of hydrocephalus in the latter two groups were prepared by intracerebroventricular injection of kaolin suspension;rats in the sham-operated group were injected with same amount of normal saline into the lateral ventricle.The p38 MAPK specific inhibitor SB203580(10 mg/kg)was intraperitoneally injected into the rats of SB group on the 8th d of modeling for 7 consecutive d;same volume of dimethylsulfoxide was given to the rats of hydrocephalus group on the 8th d of modeling for 7 consecutive d;rats in the sham-operated group did not give any treatment.The severity of hydrocephalus in these rats was observed by MRI.The inflammatory factor tumor necrosis factor(TNF)-αlevel in the cerebrospinal fluid was detected by enzyme-linked immunosorbent assay(ELISA).The AQP4 and TNF-αmRNA expressions were detected by real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR).The phosphorylated p38 MAPK and AQP4 expressions in the periventricular brain tissues were detected by Western blotting and immunohistochemistry.Results No hydrocephalus developed in sham-operated group and hydrocephalus developed in the latter two groups.As compared with sham-operated group,hydrocephalus group and SB group had significantly increased lateral ventricle volume,significantly aggravated periventricular edema,significantly higher EVAN's index,and statistically increased brain water content(P<0.05).Two weeks after modeling,the TNF-αexpression levels in cerebrospinal fluid of sham-operated group,hydrocephalus group and SB group were(20.49±0.96),(42.04±3.17),and(28.00±3.71)pg/mL,respectively,with significant differences(F=186.000,P<0.001);the TNF-αexpres

关 键 词：脑积水 P38丝裂原活化蛋白激酶 水通道蛋白4 

分 类 号：R742.7[医药卫生—神经病学与精神病学]