c-jun氨基末端激酶在弥漫性脑创伤大鼠脑中表达及对神经元自噬的影响  

Effect of expression of c-jun N-terminal kinase on neuron autophagy following diffuse brain injury in rats

作　　者：洪铭岩[1] 崔建忠[1] 李冉[2] 田艳霞[2] 王焕[2] 王海涛[2] 高俊玲[2] 

机构地区：[1]河北医科大学外科学教研室,石家庄050017 [2]河北联合大学基础医学院

出　　处：《中华外科杂志》2012年第2期166-170,共5页Chinese Journal of Surgery

基　　金：河北省自然科学基金资助项目（C2009001247）

摘　　要：目的 探讨大鼠弥漫性脑创伤后c-jun氨基末端激酶（JNK）通路的表达和对大鼠神经元自噬的影响和机制.方法选用雄性SD大鼠216只,随机分为（1）颅脑创伤组（n=54）；（2）SP600125干预组（n=54）；（3）DMSO对照组（n=54）；（4）假手术对照组（n=54）.每组又随机平均分为伤后1、6、12、24、48和72 h6个亚组.观察伤后皮质区神经细胞组织形态变化,Western blot法、免疫组化法检测顶叶皮质区p-JNK、p-P53、DRAM和Beclin-1的表达.结果 颅脑创伤组伤后6h光镜下即可见大脑皮质区神经细胞胞体收缩呈三角形,胞浆嗜色性减弱,核皱缩浓染,细胞周围出现空隙,即神经细胞变性坏死改变；SP600125干预组上述改变明显减轻,与颅脑创伤组比较DMSO对照组变化不大,假手术对照组可见神经元数量多,排列整齐,形态完整,核质均匀,核仁清晰.免疫组化与Western blot法结果显示,与SP600125干预组比较,颅脑创伤组p-JNK活性在伤后6、12和24 h显著增高（F=17.902,P＜0.05）,p-P53活性在伤后12、24、48和72 h显著增高（F=7.107,P＜0.05）,DRAM活性在伤后6、12、24、48和72 h显著增高（F=15.455,P＜0.05）和Beclin-1活性在伤后6、12、24、48和72 h显著增高（F=11.517,P＜0.05）；与颅脑创伤组比较,DMSO对照组中p-JNK、p-P53、DRAM、Beclin-1在各时间点差异均无统计学意义（F=1.509,P＞0.05）.结论 本研究表明SP600125可改善脑创伤后神经元的自噬性损伤,其机制与调节脑创伤后JNK信号活化水平有关,提示颅脑创伤后JNK通路的激活可能是调控神经元自噬的重要机制之一.Objective To study the effect and potential mechanism of expression of c-jun N-terminal kinase（JNK）signal pathway on neuron autophagy after diffuse brain injury（DBI）.Methods Male Sprague Dawley rats（n =216）were randomly divided into four groups：DBI group（n =54）,SP600125 intervene group（n=54）,DMSO group（n =54）and sham operation group（n =54）.DBI rat model was established according to the description of Marmarou DBI.At different time points（1,6,12,24,48 and 72 h）after operation,the histopathologic changes of neurons in cortex were observed by HE staining method; The expression of p-JNK,p-P53,DRAM and Beclin-1 were detected by Western blot and immunohistochemistry.Results The results showed that under light microscope degenerated and necrotic neurons were observed to be scattered in cortex at 6 h after operation in DBI group,but these changes were low in SP600125 intervene group.Compared with SP600125 intervene group,the expression of p-JNK in DBI group were enhanced obviously at 6,12 and 24 h（F =17.902,P 〈 0.05）; the expression of p-P53 in DBI group were enhanced obviously at 12,24,48 and 72 h（F =7.107,P 〈 0.05）; the expression of DRAM in DBI group were enhanced obviously at 6,12,24,48 and 72 h（F =15.455,P 〈 0.05）; the expression of Beclin-1 in DBI group were enhanced obviously at 6,12,24,48 and 72 h（F =11.517,P 〈 0.05）.Compered with DBI group,the expression of p-JNK,p-PS3,DRAM and Beclin-1 in DMSO group were similar at 1,6,12,24,48 and 72 h（F =1.509,P 〉 0.05）.Conclusions The present results indicate that SP600125 can dramatically improve trauma brain injury from autophagy after DBI and the molecular mechanism is related to the modulation of JNK signal pathway following DBI,while it measures the neuron autophagy by means of intervening JNK signal pathway.

关 键 词：脑损伤 慢性 JNK丝裂原活化蛋白激酶类 蒽类 自噬 大鼠 

分 类 号：R651.15[医药卫生—外科学]