P-JNK在A_(2A)R敲除新生小鼠HIBD细胞凋亡中的表达研究  

Expressions of P-JNK in nerve cell apoptosis of A_(2A)R knockout newborn mice after hypoxia/ischemia brain damage

作　　者：范海玲[1] 尹水贵[1] 娄普[1] 任素伟[1] 黄胜[1] 陈翔[2] 

机构地区：[1]温州医学院 [2]温州医学院附属第二医院育英儿童医院,浙江温州325027

出　　处：《中国应用生理学杂志》2013年第2期187-192,共6页Chinese Journal of Applied Physiology

基　　金：浙江省科技厅钱江人才项目(2009R10024);温州市科技局国际合作项目(H20110018)

摘　　要：目的:探讨腺苷A2A受体(A2AR)敲除的新生小鼠在缺氧缺血性脑损伤(HIBD)后海马CA1区磷酸化c-jun氨基末端激酶(P-JNK)激活的动态变化及其与神经细胞凋亡的关系。方法:参照经典Rice-Vannucci法建立7日龄新生小鼠HIBD模型。A2AR敲除(KO)小鼠及同窝野生型(WT)小鼠80只,设假手术(S)组,模型组(M)分1 d,3 d,7d,共8小组。采用三种发育反射(翻正反射、趋地反射和悬崖逃避反射)对小鼠进行短期神经功能评定,苏木精-伊红(HE)染色、尼氏染色、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)染色法观察敲除A2AR对新生小鼠HIBD后神经细胞凋亡的影响,免疫组化检测P-JNK阳性细胞的表达。结果:敲除A2AR加重HIBD模型小鼠神经功能缺损、脑组织病理学损伤,增加神经细胞凋亡;HIBD后缺血侧海马CA1区P-JNK阳性细胞的表达迅速增加,与S组比较均有显著差异(P<0.01),模型组于1 d达高峰,且在相应的时间点(1 d,3 d,7 d)KO组P-JNK阳性细胞的表达显著高于WT组(P<0.01,P<0.05,P>0.05);直线相关分析表明新生小鼠HIBD后神经细胞凋亡与P-JNK阳性细胞的表达呈显著正相关(r=0.837,P<0.01)。结论:敲除A2AR增加新生小鼠HIBD神经细胞的凋亡及加重脑损害的过程,其作用机制可能与早期P-JNK的持续激活有关。Objective： To investigate the effect of adenosine AzA receptor knockout （A2ARKO） on relationship between continuous activation of phospho-c-Jun N-tenniusl kinase （P-JNK） and expression of nerve cell apoptosis in hippocampus CA1 domain of newborn mice after hy- poxia/ischemia brain damage（HIBD） and its potential mechanism. Methods： ARKO mice and adenosine A2A receptor wildtype （A2ARWT） littermates （ n = 80） were divided into Sham operation group（S） and model group（M）, 1, 3 and 7 day after HIBD, totally 8 groups. HIBD was developed with 7 day-old neonatal mice according classical Rice-Vannucci method. It was tested the effect of A2ARKO on short-term neurofunctional outcomes consisted of three developmental reflexes （righting, geotaxis and cliff aversion）, the changes of brain pathology with hema- toxylin-eosin （HE） staining and Nissl staining, the expressiom of nerve cell apoptosis with terminal deoxynucleotidyl transferase mediated dU＇IP-biotin nick-end labeling（ TUNEL） staining and P-JNK were observed by immunohistochemislry. Results： The neurological behavior injuries and brain histopatholngical damages and nerve apoptosis cells were aggravated in ARKO newbem mice after HIBD. The positive ex- pressions of P-JNK were significantly higher in the ischemic hippocampus CA1 domain after HIBD than ones in gwup S respectively（ P 〈 0.01）, reaching to peak at 1 day and then began gradually decreasing. P-JNK expression in model knockout（MKO） at 1, 3 and 7 day increased greatly compared to those in the previous time point of corresponding model wildtypo （MWT） （ P 〈 0.01, P 〈 0.05, P 〉 0.05） ; there was a positive correlation between the expressions of P-JNK and nerve cell apoptosis after HIBD in newbem mice（ r = 0. 837, P 〈 0.0l）. C.smdusion： Early continuous activation of P-JNK might be involved in the aggravated nerve apoptosis cells and brain damage induced by A2A RKO newborn mice after HIBD.

关 键 词：腺苷A2A受体 敲除 磷酸化c-jun氨基末端激酶 细胞凋亡 缺氧缺血性脑损伤 

分 类 号：R73-3[医药卫生—肿瘤]