TAK1抑制剂5Z-7-oxozeaenol对大鼠脑外伤海马CA1区的神经保护作用研究  

TAK1 inhibitor 5Z-7-oxozeaenol attenuates traumatic brain injury in rat hippocampal CA1 region

作　　者：张顶顶[1] 杭春华[1,2,3] 胡阳春 孙青[1] 庄宗[1] 陈振瑞[2,3] 申伟[2,3] 

机构地区：[1]南京大学医学院临床学院南京军区南京总医院神经外科,江苏南京210002 [2]南方医科大学南京临床医学院 [3]南京总医院神经外科,江苏南京210002

出　　处：《中华神经外科疾病研究杂志》2013年第4期327-331,共5页Chinese Journal of Neurosurgical Disease Research

基　　金：国家自然科学基金资助项目(81171170);江苏省自然科学基金资助项目(BK2010459);军区医学科技创新课题资助项目(10Z024)

摘　　要：目的探讨转化生长因子β激活激酶(TAK1)选择性抑制剂5Z-7-oxozeaenol在创伤性颅脑损伤中对伤侧海马的保护作用及可能机制;方法 72只雄性Sprague-Dawley大鼠随机分为3组:假手术组,外伤+二甲基亚砜组,外伤+5Z-7-oxozeaenol组,按照改良的Feeney法制作大鼠颅脑外伤模型,外伤后10 min通过左侧侧脑室给药,24 h后取标本,Western blot检测TAK1和磷酸化TAK1的表达,并用凝胶迁移实验检测其下游核因子κB和活化蛋白-1的活性,采用原位末端脱氧核苷酸转移酶介导的生物素脱氧尿嘧啶核苷酸缺口末端标记法(TUNEL)检测海马CA1区神经元的凋亡情况,胶质纤维酸性蛋白(GFAP)免疫组化染色检测星形胶质细胞的表达;结果与溶剂组相比,治疗组伤侧海马TAK1和磷酸化TAK1的表达量、核因子κB和活化蛋白-1的DNA结合活性显著降低,同时海马CA1区TUNEL和GFAP阳性细胞明显减少(P<0.05)。结论 5Z-7-oxozeaenol通过抑制TAK1信号通路降低海马组织中NF-κB和AP-1的活性起到抗凋亡作用。Objective To investigate the protection of TAK1 inhibitor 5Zo7-oxozeaenol on the ipsilateral hippocampus after traumatic brain injury and its mechanism. Methods A total of 72 adult male Spragne-Dawley rats were randomly divided into control group, vehicle group and 5Z-7-oxozeaenol-treated group. Traumatic brain injury was induced using a modified Feeneyg weight-drop model. The TAK1 inhibitor 5Z-7-oxozeaenol （20 I.Lg） was injected intracerebroventricularly at 10 min post TBI. The expressions of TAK1 and phosphorylated TAK1 （p-TAK1） in the ipsilateral hippoeampus were analyzed by Western blot analysis. Neuronal apoptosis was evaluated by terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nick end labelling （TUNEL）. Nuclear factor-KB （NF-KB） and activator protein 1 （AP-1） activities were also measured by eleetrophoretic mobility shift assay （EMSA）. Immunohistochemieal staining was employed to identify activated astrocytes. Results Western blot analysis showed a significant increase of TAK1 and p-TAK1 24 h after TBI in the ipsilateral hippocampus. Intracerbroventricular injection with 5Z-7-oxozeaenol （20 μg） significantly reduced the expression of TAK1 and p-TAK1 protein 24 h after injury. TAK1 inhibition further reduced NF-KB and AP-1 activation. The TUNEL and glial fibrillary acidic protein positive cells in CA1 region were significantly decreased after the treatment with 5Z-7-oxozeaenol post TBI. Conclusion The present study demonstrates that treatment with 5Z-7-oxozeaenol prevents post- traumatic hippocampal neuronal apoptosis by reducing the activities of NF-KB and AP-1 in rat hippoeampus.

关 键 词：5Z-7-oxozeaenol 转化生长因子β激活激酶 创伤性脑损伤 凋亡 

分 类 号：R651[医药卫生—外科学]