小鼠不同频率轻型TBI后脑组织tau蛋白的毒性差异研究  

Toxicity of tau protein in brain tissue of mice with different frequencies of mild TBI

作　　者：程兴滕 尹亚东 张赛[3] 孙中磊 Cheng Xingteng;Yin Yadong;Zhang Sai;Sun Zhonglei(Tengnan Hospital of Zaozhuang Mining Group,Zaozhuang Shandong 277000,China;Central Hospital of Zaozhuang Mining Group,Zaozhuang Shandong 277100,China;Institute of Brain Science,Affiliated Hospital of Armed Police Logistics College,Tianjin 300162,China)

机构地区：[1]枣庄矿业集团滕南医院,山东枣庄277000 [2]枣矿集团中心医院,山东枣庄277100 [3]武警后勤学院附属医院脑科研究所,天津300162

出　　处：《遵义医科大学学报》2021年第2期159-164,共6页Journal of Zunyi Medical University

基　　金：国家重点研发计划项目(NO:2016YFC1101503)。

摘　　要：目的探索不同频率轻型颅脑创伤(Traumatic brain injury,TBI)后脑组织tau蛋白的不同及其毒性的差异。方法小鼠TBI模型制备成功后取脑组织,通过快速蛋白液相色谱(FPLC)法分析并通过免疫沉淀提取可溶性tau低聚物,采用Western blot法分析tau低聚物的结构特点,MTT和乳酸脱氢酶法测定tau低聚物的毒性,最后分离提取突触小体检测tau低聚物对突触小体的影响。结果Western blot抗(tau-5、T22、TOMA1)检测结果RB-3周组均与sham组有明显差异(P<0.05);FPLC色谱图分析表明SB-24 h、SB-3周、sham组与RB-3周组tau低聚物峰值存在差异;tau低聚物与SH-SY5Y、原代神经元共培养结果RB-3周和4R-Tau组的细胞毒性增强,细胞生存率的下降(P<0.05),而T22抗体预孵育SH-SY5Y可以显著逆转RB-3周组细胞生存率下降(P<0.05);与未处理组相比突触提取物突触前标记物(Synaptophysin)水平在RB-3周和4R-Tau组降低,突触后标记物(PSD95)水平在TBI各组均降低,突触树突标记物(Drebrin)水平仅在RB-3周组降低(P<0.05),而与sham相比,仅RB-3周组可引起PSD95和Drebrin水平降低(P<0.05)。结论本研究证明,不同频率轻型TBI诱导的tau低聚物不同,表现出不同的形态特性和毒性。Objective To explore the difference of tau protein and its toxicity in brain tissue after different frequencies of mild traumatic brain injury(TBI).Methods After TBI model was successfully established in mice,the brain tissue was taken,and the soluble tau oligomer was analyzed and extracted by fast protein liquid chromatography(FPLC).The structural characteristics of tau oligomer were analyzed by Western blot.The toxicity of tau oligomer was determined by MTT and lactate dehydrogenase.Finally,the synaptosomes were isolated and extracted to detect the effect of tau oligomer on synaptosomes.Results Western blot(tau-5,T22,toma1)results showed that there were significant differences between RB-3W group and sham group(P<0.05);FPLC chromatogram analysis showed that there were differences in the peak value of tau oligomer between SB-24 h,SB-3W,sham group and RB-3W group;the results of co-culture of tau oligomer with SH-SY5Y and primary neurons showed higher cytotoxicity with RB-3W and 4R-Tau.Compared with the untreated group,pre-incubation of T22 antibody with SH-SY5Y cells could significantly reverse the decrease of cell survival rate in RB-3W group(P<0.05).Compared with the untreated group,synaptophysin levels in RB-3W and 4R-Tau groups were significantly lower.Compared with sham group,the level of PSD95 and Drebrin decreased only in RB-3W group(P<0.05).Conclusion Tau oligomers induced by different frequencies of TBI are different,showing different morphological characteristics and toxicity.

关 键 词：颅脑损伤 TAU 毒性 突触小体 

分 类 号：R615.15[医药卫生—外科学]