JNK信号通路在异氟醚诱导新生大鼠海马神经细胞凋亡的作用  

The effects of JNK pathway on isoflurane induced neuronal apoptosis in the hippocampi of neonatal rats

作　　者：沈智文[1] 韩雪[1] 李玉娟[1] 胡楚文[1] 廖朝霞[1] 柳垂亮 

机构地区：[1]中山大学孙逸仙纪念医院麻醉科,广州510120 [2]广东省佛山禅城中心医院麻醉科

出　　处：《中华行为医学与脑科学杂志》2013年第8期673-676,共4页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：广东省自然基金（$2011010004558）;国家自然科学基金（30700787/C03030301）

摘　　要：目的 探讨c-Jun氨基末端激酶（the c-Jun N-terminal kinase,JNK）信号通路对异氟醚诱导新生大鼠海马神经细胞凋亡以及对磷酸化JNK、Bcl-2和Bax蛋白表达的影响.方法 48只出生后7d（Py7）的新生大鼠按照随机数的方法随机均分为DMSO对照组（D组）、JNK抑制剂SP600125对照组（SP30组）、异氟醚＋DMSO组（Iso＋D组）和异氟醚＋SP600125组（Iso＋ SP30组）.对照组吸入空气,异氟醚组吸入1.1％异氟醚4h.麻醉前20 min,幼鼠根据分组分别侧脑室注射SP600125 30 μg或者12％ DMS0 5μl.麻醉结束后6h,部分幼鼠灌注取脑,TUNEL荧光染色检测脑海马CA1区神经细胞凋亡（n=6）；部分幼鼠取新鲜脑皮质,Western blotting法检测磷酸化JNK、Bcl-2和Bax蛋白表达的变化（n=6）.组间资料的比较采用单因素方差分析.结果 幼鼠海马CA1区的TUNEL阳性细胞数比较,Iso＋D组[（135.72±21.26）个/mm2]比D组[（24.07±1.35）个/mm2]增加5倍（P＜0.01）；与Iso＋D组相比,Iso＋ SP30组[（42.49±5.56）个/mm2] CA1区的TUNEL阳性细胞数降低了84％ （P＜0.05）.Iso＋D组磷酸化JNK P46表达比D组增加44.1％ （P＜0.01）,而Iso＋ SP30组显著降低了磷酸化JNK P54 （p＜ 0.05）和P46（P＜ 0.01）的表达.Iso＋D组Bax表达比D组增加1.5倍（P＜0.05）,Bcl-2蛋白表达比D组下降42.2％ （P＜0.05）；而Iso＋ SP30组Bax表达下降（P＜0.05）,Bcl-2蛋白表达上调（P＜0.01）.D组与SP30组TUNEL阳性细胞数,磷酸化JNK,Bcl-2和Bax蛋白表达差异均无统计学意义.结论 JNK信号通路激活促进了异氟醚诱导发育期大鼠海马神经细胞凋亡,SP600125通过维持Bcl-2,降低Bax表达产生抑制凋亡作用.Objective To investigate the effects of the c-Jun N-terminal kinase （JNK）pathway on isoflurane induced neuronal apoptosis and the proteins expression of phospho-JNK,Bcl-2 and Bax in the hippocampi of neonatal rats.Methods Forty-eight neonatal rats at postnatal day 7 （P7） were randomly assigned into 4 groups：DMSO control group （group D）,SP600125 control group （group SP30）,isoflurane ＋ DMSO group （group Iso ＋D）,isoflurane ＋ SP600125 group （group Iso ＋ SP30）.Rats were exposed to air （control group） or 1.1% isoflurane （isoflurane group） for 4 h.The JNK inhibitor SP600125 at 30 μg or 12% DMSO 5 μl was intraventricularly administered 20 min before the exposure.The brains of some rats in each group were perfused and embedded by paraffin 6 h after the exposure.Neuronal apoptosis in the hippocampi CA1 area was detected by TUNEL （n =6）.The fresh hippocampi of other rats in each group were dissected 6 h after the exposure and the proteins expression of phospho-JNK,Bcl-2 and Bax were detected by Western blot （n =6）.One way ANOVA were used for data analysis among groups.Results The number of TUNEL positive cells in the hippocampal CA1 regions in group Iso ＋D （135.72 ±21.26 per mm2） increased by 5 folds compared with group D （24.07 ± 1.35 per mm2） （P〈0.01） ;while the number of apoptotic cells in group Iso ＋ SP30 （42.49 ± 5.56 per mm2） decreased by 84% （P 〈 0.05）compared with group Iso ＋ D.The expression of phospho-JNK p46 kd in group Iso ＋ D increased by 44.1% （P 〈0.01）,while both phospho-JNK at p46kd and at p54kd in group Iso ＋ SP30 decreased significantly （P〈0.05,P 〈0.01） compared with group Iso ＋ D.The protein expression of Bax increased 1.5 folds （P〈0.05） and Bcl-2 decreased by 42.2% （P〈0.05） in group Iso ＋ D compared to group D;while SP600125 significantly decreased expression of Bax （P 〈0.05） and increased expression of Bcl-2 （P〈0.01）.Conclusion JNK activation contributes to isoflurane-induce

关 键 词：细胞凋亡 异氟醚 c—Jun氨基端激酶 海马 

分 类 号：R614[医药卫生—麻醉学]