摘要
目的探讨急性创伤性颅脑损伤(TBI)后突触形成相关蛋白突触关联蛋白(Snap)25、神经突触素(Synapsin)1和线粒体功能异常相关蛋白单胺氧化酶B(Maob)、NADH脱氢酶(辅酶Q)Fe-S蛋白(Ndufs)2在大鼠海马组织中的表达及其意义。方法采用大鼠可控性皮质打击伤(CCI)建立中度TBI模型,CCI后48 h(CCI组,n=13)为急性期观察点;假手术组(n=13)与CCI组处理步骤一致,仅不打击皮质。采用超高效液相色谱和同位素标记相对与绝对定量(iTRAQ)蛋白质质谱分析CCI组(n=5)与假手术组(n=5)海马组织之间的差异蛋白。采用生物信息学方法分析差异蛋白的细胞组分、生物进程和分子功能,并进行经典通路富集分析及疾病功能富集分析。采用透射电子显微镜观察两组海马组织神经元核和线粒体的超微结构变化。采用蛋白质免疫印迹法(WB)检测Snap25、Synapsin1、Maob以及Ndufs2蛋白的表达。结果蛋白质质谱数据结果显示,与假手术组相比,CCI组上调的差异蛋白共55种(差异倍数≥1.50),下调的差异蛋白共110种(差异倍数≤0.65)。生物信息学分析结果显示,富集的经典通路包括突触形成信号通路[-log_(10)(P值)=7.29]和线粒体功能障碍信号通路[-log_(10)(P值)=4.06]等;突触形成信号通路中富集了Snap25和Synapsin1等蛋白;线粒体功能障碍信号通路中富集了Maob和Ndufs2等蛋白。透射电子显微镜观察发现,CCI组出现神经元核固缩、线粒体嵴断裂。WB检测结果显示,与假手术组(n=5)比较,CCI组(n=5)Snap25(分别为0.594±0.025、1.000±0.141,t=4.92)、Synapsin1(分别为0.597±0.077、1.000±0.100,t=5.56)、Maob(分别为0.528±0.042、1.000±0.160,t=4.94)以及Ndufs2(分别为0.549±0.057、1.000±0.131,t=5.45)蛋白的相对表达量均下降,差异均具有统计学意义(均P<0.01)。结论突触形成相关蛋白Snap25、Synapsin1和线粒体功能异常相关蛋白Maob、Ndufs2在急性TBI大鼠海马组织中的表达下降,可能参与TBI的�
Objective To investigate the expression and significance of synaptogenesis related proteins synaptosomal-associated protein(Snap)25 and Synapsinl and mitochondrial dysfunetion related proteins monoamine oxidase B(Maob)and NADH dehydrogenase(ubiquinone)Fe-S protein 2(Ndufs2)in hippocampus of rats with acute traumatic brain injury(TBI).Methods The moderate TBI model was established by applying controlled cortical injury(CCI)to rats.The time point of 48 h after CCI(CCI group,n=13)was set for observation in the acute phase.The sham group(n=13)underwent the same procedure without percussion.The expression of differential proteins between CCI group(n=5)and sharm group(n=5)was analyzed by ultra-high performance liquid chromatography and isotope labeled relative and absolute quantitative(iTRAQ)protein mass spectrometry.Bioinformatics methods were employed to analyze the cellular components,biological processes and molecular functions of differential proteins,and classical pathway enrichment analysis and disease function enrichment analysis were carried out.The ultrastructural changes of neuronal nuclei and mitochondria in the hippocampus of the two groups were observed by transmission electron microscopy.The expression of Snap25,Synapsinl,Maob and Ndufs2 proteins were detected by Western blot(WB).Results The results of protein mass spectrometry showed that compared with the sham group,55 differential proteins were up-regulated in CCI group(fold change≥1.50)and 110 differential proteins were down-regulated(fold change≤0.65).Bioinformatics analysis showed that the enriched classical pathways included synaptogenesis signaling pathway[-log_(10)(P value)=7.29]and mitochondrial dysfunction signaling pathway[-log_(10)(P value)=4.06];Snap25 and Synapsinl were enriched in synaptogenesis signaling pathway;Maob and Ndufs2 proteins were enriched in mitochondrial dysfunction signaling pathway.Electron microscopy demonstrated that the CCI group had pyknosis of neuron nucleus and rupture of mitochondrial crista.WB results showed th
作者
牛非
冯洁
茆翔
张斌
徐晓健
刘佰运
Niu Fei;Feng Jie;Mao Xiang;Zhang Bin;Xu Xiojian;Liu Baiyun(Beijing Neurosurgical Institute,Capial Medical University,Beijing 100070,China;Department of Neurosurgery,the First Affiliated Hospital of Anhui Medical University,Hefei 230022,China;Department of Critical Care Medicine,Beijing Tiantan Hospital,Capital Medical University,Bejing 100070,China;Neurosurgry Center,Beijing Tiantan Hospital,Capital Medical University,Bejing 100070,China)
出处
《中华神经外科杂志》
CSCD
北大核心
2023年第6期612-617,共6页
Chinese Journal of Neurosurgery
基金
国家自然科学基金(81771327)
北京市卫生健康委员会2022年预算项目(11000022T000000423832)
北京市神经外科研究所科研培育基金。
