创伤性脑损伤后tau蛋白磷酸化水平动态变化及意义  

Dynamic change and significance of tau protein phosphorylation after traumatic brain injury

作　　者：赵子艾 宁亚蕾[1] 杨楠[1] 彭艳 熊仁平[1] 陈惺 白伟 周元国[1] 

机构地区：[1]第三军医大学附属大坪医院野战外科研究所分子生物学中心,创伤、烧伤与复合伤国家重点实验室,重庆400042

出　　处：《中华创伤杂志》2014年第4期344-348,共5页Chinese Journal of Trauma

基　　金：国家自然科学基金资助项目（31171022,81201461）;重庆市自然科学基金资助项目（cstc2012jjA10107）

摘　　要：目的研究小鼠创伤性脑损伤（traumatic brain injury，TBI）后急性期、亚急性期及慢性期tau蛋白磷酸化水平动态变化规律。方法采用可控皮质打击方法建立小鼠中度TBI模型，无特定病原体（specific pathogen free，SPF）级C57BL／6雄性小鼠40只按随机数字表法分为创伤组、假手术组，创伤组又分为3个亚组：24h、7d及1个月组，每组10只。采用免疫组化方法检测伤后24h、7d、1个月脑组织tau蛋白磷酸化水平变化、胶质细胞增生程度、神经元丢失及淀粉样肽前体蛋白（amyloid precursor protein，APP）标记的轴突损伤。结果创伤24h、7d、1个月组对侧海马齿状回区tau蛋白Set404位点磷酸化水平分别为4451．021±471．731，5339．826±794．071，1906．0364-369．808，均较假手术组368．6304-231．038明显升高（f=23．316，18．033，9．979，P〈0．01）。而其余Thr205、Ser262位点磷酸化水平未见明显改变。胶质纤维酸性蛋白（glial fibrous acid protein，GFAP）免疫组化染色结果显示，创伤24h、7d组吸光度（4）值分别为30495．233±6174．094，86427．920±11896．771，均显著高于假手术组6825．653±3451．641（t=9．573，9．573，P〈0．01），而创伤1个月组9511．471±5431．077与假手术组比较差异无统计学意义（t=1．181，P〉0．05）。APP免疫组化结果显示，假手术组、创伤24h、7d及1个月组A值分别为459．0614±368．608，1802．6954±491．938，2762．852±636．524，2333．118±511．731，创伤组均高于假手术组（t=5．684，7．960，7．697，P〈0．01）。各时相点NeuN标记神经元计数结果显示，对侧齿状回区神经元在创伤24h组与假手术组[（290．056±39．162）：（314．019±42．397）]差异无统计学意义（t=-1．017，P〉0．05），创伤7d、1个月组分别为250．509±17．592，242．717±17．045，均少于假手术组（t=-3．112，-3．506，P〈0．01）。结论TBI后tau蛋白Ser404位点Objective To investigate the dynamic changes in phosphorylation levels of tau proteins in the acute, sub-acute and chronic stages after traumatic brain injury （TBI） in mice. Methods Mouse models with moderate TBI were developed using the controlled cortical impactor. Forty specific pathogen free （SPF） male C57BL/6 mice were allocated to TBI group and sham operation group according to the random number table. TBI group was subdivided at 24 hours, 7 days, and 1 month. There were 10 mice per group. Levels of tau protein phosphorylation in the brain, gliosis, neuronal loss, and amyloidprecursor protein （APP）-labeled axonal injury were detected by immunohistochemistry at 24 hours, 7 days, and 1 month after TBI. Results Levels of tau protein phosphorylation in contralateral dentate gyrus at Ser404 site were4 451.021±471.731, 5 339. 826±794.071, and 1 906.036±369. 808 at 24 hours, 7 days, and 1 month posttrauma, far higher than 368. 630±231. 038 in sham operation group （ t = 23. 316, 18. 033, 9. 979 respectively, P 〈 0.01 ）. Whereas, there were no significant differences of tau protein phosphorylation at Ser262 and Thr205 between TBI and sham operation groups at all time points. Immunohistochemical staining for glial fibrous acid protein （GFAP） showed the absorbance value in TBI group was 30 495. 233±6 174. 094 and 86 427. 920±11 896.771 at 24 hours and 7 days, far higher than 6 825. 653±3 451. 641 in sham operation group （ t = 9.573, 9.573 respectively, P 〈 0.01 ）. Besides, the absorbance value in TBI group was 9 511. 471±5 431. 077 at 1 month, with no statistical difference in contrast with sham operation group （ t = 1. 181, P 〉 0. 05 ）. Immunohistochemical staining for APP indicated the absorbance value in TBI group was 1 802. 695±491. 938, 2 762. 852±636. 524, and 2 333. 118±511.731 at 24 hours, 7 days, and 1 month, higher than 459. 061±368. 608 in sham operation group （ t = 5. 684, 7. 960, 7. 697 respectively, P 〈 0. 01 ）. Number of NeuN-labeled neurons in contral

关 键 词：脑损伤 TAU蛋白质类 认知功能障碍 

分 类 号：R651.1[医药卫生—外科学]