c-Jun氨基端激酶通路抑制剂SP600125对大鼠脑缺血再灌注后自噬的影响  

Effects of inhibits c-Jun N-terminal kinase SP600125 on autophagy after cerebral ischemia/reperfusion in rats

作　　者：江秀龙[1] 雷惠新[1] 汪银洲[1] 张旭[1] JIANG Xiu - long;LEI Hui - xin;WANG Yin - zhou;ZHANG Xu(Department of Neurology, Fujian Provincial Hospita;Fufian Medical University Sheng Li Clinical Medical College ,Fuzhou 350001, Chin)

机构地区：[1]福建医科大学省立临床学院福建省立医院神经内科,福州350001

出　　处：《中国临床药理学杂志》2018年第12期1432-1435,共4页The Chinese Journal of Clinical Pharmacology

基　　金：福建省自然科学基金面上基金项目资助(2015J01371)

摘　　要：目的研究c-Jun氨基端激酶(JNK)特异性抑制SP600125对大鼠脑缺血再灌注后自噬性蛋白Beclin1、髓样细胞白血病基因-1(Mcl-1)蛋白和B淋巴瘤-2基因相关X(Bax)蛋白表达的影响。方法按照体重将SD大鼠随机分为3组,每组10只:假手术组、模型组和实验组。模型组和实验组均采用线拴法制作SD大鼠大脑中动脉阻塞再灌注(MCAO)模型,脑缺血1 h再灌注72 h。造模前,实验组先向大鼠侧脑室内注射3μg·μL^(-1)SP600125溶液10μL,1 h后进行MCAO制模。以组织免疫荧光染色方法测定MCAO后JNK与Beclin1的表达;以免疫印迹法测定MCAO后JNK、Beclin1、Mcl-1和Bax蛋白表达。结果给药后,假手术组、模型组、实验组的JNK条带光密度值分别是0.11±0.05,0.83±0.03,0.56±0.04;这3组的Beclin1条带光密度值分别为0.14±0.04,1.77±0.05,0.86±0.05;这3组的Mcl-1条带光密度值分别为0.17±0.03,0.79±0.04,1.46±0.05;这3组的Bax条带光密度值分别为0.26±0.05,1.15±0.06,0.47±0.04。模型组与假手术组比较,JNK、Beclin1、Mcl-1和Bax值均明显增高,差异均有统计学意义(均P<0.01);实验组与模型组比较,JNK、Beclin1和Bax值均明显下降而Mcl-1值明显升高,差异均有统计学意义(均P<0.01)。结论通过抑制JNK可下调脑缺血再灌注后Bax与Beclin1的表达、上调Mcl-1蛋白表达,推测JNK可能参与调节缺血再灌注后神经元的自噬。Objective To investigate the effects of inhibits c-Jun N-terminal kinase（JNK）activation SP600125 on the expression of Beclin1,myeloid cell leukemia-1（Mcl-1）,and BCL 2-associated X（Bax）after cerebral ischemia/reperfusion in rats.Methods SD rats were randomly assigned into 3 groups：sham-operated group,experimental group and model group,each group had 10 rats.The middle cerebral artery occlusion（MCAO）was induced using the intraluminal suture occlusion technique in experimental group and model group.Reperfusion of cerebral blood flow was allowed by gently removing the monofilament after 1 h ischemia,followed by 72 h reperfusion.Concentration of 3μg·μL^-1SP600125 10μL were infused stereotactically into the ipsilateral hemispheric region 1 h before MCAO in the experimental group.The expressions of JNK,Beclin1 in the brain were detected by immunofluorescent labeling.The expressions of JNK,Beclin1,Mcl-1,Bax proteins were detected by Western blot.Results After treatment,the JNK gray values respectively in the sham group,the model group and the experimental group were 0.11±0.05,0.83±0.03,0.56±0.04;the Beclin1 gray values respectively in the 3 groups were0.14±0.04,1.77±0.05,0.86±0.05;the Mcl-1 gray values respectively in the 3 groups were 0.17±0.03,0.79±0.04,1.46±0.05;the gray Bax values respectively in the 3 groups were 0.26±0.05,1.15±0.06,0.47±0.04.Compared with the sham group,increased values of JNK,Beclin1,Mcl-1 and Bax were observed in the model group,the difference was statistically significant（all P〈0.01）.Compared with the model group,reduced values of JNK,Beclin1,Bax and increased values of Mcl-1 protein in the experimental group,the difference was statistically significant（all P〈0.01）.Conclusion The expression of Beclin1 and Bax could be decreased and Mcl-1 protein could be upregulated by anti-JNK.The JNK potentially modulates post-ischemia neuronal autophagy.

关 键 词：脑缺血 C-JUN氨基端激酶 Beclin1蛋白 髓样细胞白血病基因-1蛋白 B淋巴瘤-2基因相关X蛋白 

分 类 号：R97[医药卫生—药品]