依达拉奉对Aβ_(25-35)诱导的PC12细胞MAPKs信号通路的影响  

The Effect of Edaravone on MAPKs Signal Pathway Associated with Aβ_(25-35) Treatment in PC12 Cells

作　　者：张桂莲[1] 郭英英[1] 张磊[2] 李婷婷[1] 杜赟[1] 姚丽[1] 张王刚[3] 吴海琴[1] 马珠琳 

机构地区：[1]西安交通大学第二附属医院神经内科,西安710004 [2]陕西中医学院第二附属医院神经内科,咸阳712000 [3]西安交通大学第二附属医院血液内科,西安710004

出　　处：《四川大学学报（医学版）》2015年第2期179-184,190,共7页Journal of Sichuan University(Medical Sciences)

基　　金：陕西省科技攻关项目(No.2010K16-08-02);西安交通大学第二附属医院人才基金项目〔No.RC(GG)201409〕资助

摘　　要：目的探讨自由基清除剂依达拉奉通过丝裂原活化蛋白激酶(MAPKs)信号转导通路发挥细胞保护作用的途径,为阿尔茨海默病(AD)发病机制及AD治疗新药的研发增添理论依据。方法培养肾上腺嗜铬瘤细胞(PC12细胞),根据不同的干预措施将细胞分为4组。阴性对照组(高糖DMEM组);AD模型组(30μmol/L Aβ25-35处理组);抑制剂对照组:10μmol/L SB203580〔丝裂原活化蛋白激酶p38(p38)抑制剂〕、10μmol/L SP600125〔c-Jun氨基末端激酶(JNK)抑制剂〕或10μmol/L PD98059〔细胞外调节蛋白激酶(ERK)抑制剂〕处理组;依达拉奉低、中、高剂量组:依达拉奉20、40、80μmol/L及30μmol/L Aβ25-35共同孵育组。Western blot检测各组细胞p-p38、p-JNK及p-ERK蛋白的表达;RT-PCR法检测以上各组(抑制剂对照组仅加入SB203580 10μmol/L)p38 mRNA的表达。结果 1与阴性对照组比,AD模型组p-p38蛋白表达增高(P<0.01);与AD模型组比,抑制剂对照组和依达拉奉各组p-p38蛋白的表达减低(P<0.05);依达拉奉3组的p-p38蛋白表达高于抑制剂对照组(P<0.05),与低剂量依达拉奉组比,中剂量依达拉奉组p-p38蛋白有所减低(P<0.05)。2与阴性对照组比,AD模型组p-JNK蛋白表达增高(P<0.05);与AD模型组比,抑制剂对照组p-JNK蛋白的表达减低(P<0.05)。3 pERK表达在各组之间差异无统计学意义(P>0.05)。4与阴性对照组比,AD模型组的p38mRNA表达增加,而抑制剂对照组的p38mRNA表达减少(分别P<0.05);中、高剂量依达拉奉组p38mRNA表达比AD模型组减低,且比抑制剂对照组还低(P<0.05);以40μmol/L依达拉奉组p38mRNA表达最低,与20μmol/L、80μmol/L依达拉奉组均明显不同(P<0.05)。结论 Aβ25-35可能直接激活MAPK信号通路,尤其p38及JNK,损伤PC12细胞。依达拉奉可能同时在mRNA水平及蛋白质水平,阻断p38信号转导通路,发挥抗Aβ25-35引起的PC12细胞损伤作用,有望成为治疗AD的新药。Objective To explore whether edaravone protects cells damage via mitogen-activated protein kinases（MAPKs）signal pathway,and which procedure of p38 be affected so as to add theories for AD pathogenesis and treatments.Methods According to different drugs treated,PC12 cells in vitro were divided into four groups.Negative control group：cells were treated with media alone.AD model group：cells were treated with 30μmol/L Aβ25-35.Inhibitor control group：cells were treated with 10μmol/L SB203580〔p38mitogen-activated protein kinase（p38）inhibitor〕,10μmol/L SP600125 〔c-Jun NH2 terminal kinase（JNK）inhibitor〕,or 10μmol/L PD98059 extracelular signal regulated kinase（ERK）inhibitor〕.Low-dose,middle-dose and high-dose edaravone group：cells plated for 24 hours treated with 30μmol/L Aβ25-35 and co-treated with 20,40,80μmol/L edaravone 3hours,respectively.The morphology of the treated cells were observed,the p-p38,p-JNK and p-ERK proteins in each group were tested by the Western blot.The p38 mRNA were tested in each group above（only add SB20358010μmol/L in third group）by the real time PCR.Results 1 The p-p38 protein was significantly increased in model control group compared with that in negative control group（P〈0.05）.The p-p38 protein in the inhibitor group and edaravone groups was decreased significantly（P〈0.05）when compared with that in model control group.The p-p38 proteins were significantly increased in the three edaravone groups compared with that in inhibiter control group（P〈0.05）.The p-p38 protein in middle-dose edaravone group was decreased compared with that in low-dose edaravone group（P〈0.05）.There was no relationship in dose-dependent manner about edaravone.Compared with three edaravone groups,the p-p38 protein was lower than it in high-dose edaravone inhibiter group（P〈0.05）.2 The p-JNK protein was significantly increased in model control group compared with that in negative control group（P〈0.05）.The p-JNK protein in the inhibitor group wa

关 键 词：阿尔茨海默病 Β淀粉样蛋白 依达拉奉 MAPKS PC12细胞 

分 类 号：R749.16[医药卫生—神经病学与精神病学]