癫痫幼鼠海马丝裂酶原蛋白活化激酶表达的特点及其对巨噬细胞炎性蛋白-α/趋化因子受体5表达的影响  

Express characteristics of mitogen-actived protein kinases at hippocampus in immature rats with epilepsy and its influence on the expression of macrophage inflammatory protein-1α/chemokine receptor 5 axis

作　　者：王静茹[1] 朱晓波[1] 马宇[1] 陈少杰[1] 王一彪[1] 

机构地区：[1]山东大学第二医院儿内科,济南250033

出　　处：《临床神经病学杂志》2017年第2期124-129,共6页Journal of Clinical Neurology

基　　金：山东省医药卫生科技发展计划项目(2014WSO154)

摘　　要：目的探讨癫痫幼鼠海马丝裂酶原蛋白活化激酶(MAPKs)表达的特点及其对巨噬细胞炎性蛋白-α(MIP-1α)/趋化因子受体5(CCR5)表达的影响。方法应用立体定向技术对侧脑室内注射海人酸(KA),建立幼鼠惊厥模型。将出生21 d的Wistar幼鼠分为空白对照组、磷酸盐缓冲液(PBS)对照组及KA 4h、8 h、16 h、24 h、3 d组,比较各组MAPKs表达的差异。另将出生21 d的幼鼠分为PBS+二甲亚砜(DMSO)组、KA+DMSO组、KA+PD98059(ERK1/2抑制剂)组和KA+SB203508(p38MAPK抑制剂)组,比较各组MIP-1α、CCR5表达的差异。采用Western Blot方法检测MAPKs蛋白水平及CCR5的表达。采用ELISA方法检测MIP-1α的表达。采用免疫组化染色方法检测各组大鼠海马P-ERK1/2、P-p38MAPKs等蛋白的表达。采用免疫荧光双标染色探讨P-ERK1/2和P-p38MAPK的胶质细胞来源。结果与空白对照组比较,KA 4 h、8h、16 h、24 h、3 d组P-ERK1/2水平明显增高;KA 8 h、16 h、24 h、3 d组P-P38MAPK水平明显增高(均P<0.05)。侧脑室注射KA后大鼠P-ERK1/2与P-P38MAPK表达主要分布在齿状回门区和锥体细胞层等神经元损伤明显的海马组织中。在侧脑室注射KA后,部分P-ERK1/2来源于活化的小胶质细胞及星形胶质细胞,而P-p38MAPK仅在小胶质细胞中出现免疫活性表达。与PBS+DMSO组比较,KA+DMSO组、KA+PD98059组和KA+SB203508组大鼠海马组织中MIP-1α、CCR5蛋白水平均明显增高,OX-42阳性细胞数明显增加(均P<0.05)。与KA+DMSO组比较,KA+PD98059组、KA+SB203508组大鼠海马组织中MIP-1α、CCR5蛋白水平均明显降低,OX-42阳性细胞数明显减少(均P<0.05)。结论在幼鼠癫痫发生的早期阶段,MAPKs(ERK1/2、p38MAPK)可部分地调控MIP-1α/CCR5的表达。Objective To investigate the express characteristics of mitogen-actived protein kinases （MAPKs） at hippocampus in immature rats with epilepsy and its influence on the expression of macrophage inflammatory protein-1α （MIP-1α）/chemokine receptor 5 （CCR5） axis. Methods Seizures were induced by intracerebroventricular injection of Kainic acid （KA） with stereotaxic apparatus. Wistar rats （bron 21 d） were randomly divided into blank control group, phosphate buffer （PBS） control group and KA 4 h, 8 h, 16 h, 24 h, 3 d groups. The differences of MAPKs in hippocampus among these groups were compared. Wistar rats （bron 21 d） were divided into PBS ＋ dimethyl sulfoxide （DMSO） group, KA ＋ DMSO group, KA ＋ PD98059 （ERK1/2 inhibitor） group and KA ＋ SB203508 （p38MAPK inhibitor） group. The effects of MAPKs on the expression of MIP-1α/CCR5 in hippocampus were detected. Western Blot was used to detect the level of MAPKs and CCR5 protein in each group. ELISA was used to quantify the MIP-1α protein in each group. Immunohistochemistry was performed to examine the expression of P- ERK1/2 and P-p38MAPK in each group. Results Compared with blank control group, the levels of P-ERK1/2 in KA 4 h, 8 h, 16 h, 24 h and 3 d groups were significantly increased, and the levels of P-P38MAPK in KA 8 h, 16 h , 2 4 h and 3 d groups were significantly increased （ all P 〈 0. 0 5 ） . After KA treatment , the experssion of P - ERK 1/2 and P-P38MAPK in rats were mainly distributed in the hilus of the dentate gyrus and the pyramidal cell layer,obviously damaged in hippocampus. After KA treatment, part of P-ERKI/2 was derived from activated microglia and astrocytes, while P-p38MAPK only showed the immunoreactivity in microglia. Compared with PBS ＋ DMSO group, the levels of MIP-1, CCR5 and the number of OX-d2 positive cells in hippocampus of KA ＋ DMSO group, KA ＋ PD98059 group and KA ＋ SB203508 group were significantly increased （ all P 〈 0. 05 ）. Compared with KA ＋ DMSO group, the leve

关 键 词：幼鼠 癫痫发生 丝裂原活化蛋白激酶 巨噬细胞炎性蛋白-1Α 趋化因子受体5 

分 类 号：R742.1[医药卫生—神经病学与精神病学]