慢性创伤性脑病大鼠认知与情感行为异常的分子标记物评价指标研究

The multiple cerebral concussion (MCC) frequently happen in youngman or athlete，military veterans of experienced blast injuries and victims of domestic abuse. It often leave behind more severe and intractable secondary impact syndrome(SIS)，Post-concussion Syndrome（PCS）and chronic trauma encephalopathy(CTE). The both behavior dysfunction of cognition and emotion result from CTE are often sequelae, but its mechanism is still unknowen.Especially CTE is a progressive disease that is often a long-term consequence of repetitive closed head injuries for which there is no treatment and no definitive pre-mortem diagnosis. This study will utilize the simple pendulum closed head injury blow device and the cerebral concussion(CC) discriminant regression equation for judged mild or moderate trauma brain injury to establish the CTE rats model in different impact blows,which the mathods above mentioned had previously developed by our research team. This proposal is integrating to employ the following more sensitive technology to stdy CTE in vivo for long-terms after injury,that is,methods of cognitive and emotional behavior in rats，magnetic resonance diffusion tensor imaging ( DTI ) and magnetic resonance spectroscopic imaging( MRS )，protein chip（PC） combined with surface enhanced laser desorption ionization time of flight mass spectrometry ( SELDI-TOF-MS )，neuronal and glial cell specific marker staining and stereological Technology for cell quantitative were combined used for the first time，to conduct a series of dynamic observation on the early and long-term cognitive and emotion behavior of CTE rats，molecular imaging changes of white matter fiber and cerebral molecular metabolism，differential proteins ( DP )molecular markers and quantitative pathological change of related brain tissues and peripheral blood，in order to build a objective evaluation criteria system of the behavior, molecular imaging, differential proteomic markers from the early to 18 months post-injury. To build a system biology research platform in CTE rats model for solving legal-medical problems of objective evaluation of CTE patients in early and long-term post-injury and rehabilitation intervention of translational medicine in future.

多次性脑震荡（MCC）及其慢性创伤性脑病（CTE）多发于年轻运动员、战场老兵及家庭虐待受害者，正成为社会公共卫生问题。其机制不明，缺乏客观评价鉴定指标体系。本研究用先前课题组创建的大鼠脑震荡判类回归方程与MCC大鼠复制方法，建立CTE大鼠模型。首次联合应用更敏感的大鼠认知与情感行为学方法、磁共振弥散张量成像（DTI）和磁共振频谱成像（MRS）、蛋白质芯片（PC）联合表面加强激光解析电离飞行时间质谱、神经与胶质细胞特异标记免疫染色与体视学形态定量等技术，对伤后CTE大鼠近、远期（18个月）认知与情感行为、大脑分子代谢与白质纤维在体分子影像变化，相关脑区组织与外周血差异蛋白质组标记分子及其定量病理形态改变进行动态观测研究，以构建脑损伤后早期与远期的行为学、分子影像学、差异蛋白质组分子标记三层面的在体客观评价标准实验体系。为法医解决客观评价早、晚期CTE患者，临床康复干预等转化医学问题奠定基础。

多次性脑震荡（MCC）及其后遗症慢性创伤性脑病（CTE）正在成为社会公共卫生问题，其机制不明，缺乏客观诊断指标。本研究应用课题组原创的MCC鼠复制方法与大鼠脑损伤程度昏迷判类回归方程控制入组动物，首次创建了CTE大鼠模型，并于伤后近中期（3个月内）、远期（6-12个月内）6个时间点，从三个领域6种行为学（涵盖空间认知、焦虑与抑郁情感、平衡与运动行为）观测、大脑磁共振MRS与DTI的10余个参数检测、脑额叶皮质与外周血iTRAQ蛋白组学分析、大脑组织神经元与各种胶质细胞形态学观测等四个层面开展了系统生物学动态对比观测研究。结果发现：（1）随着打击次数的增加，CTE大鼠的上述系列生物学变化，产生了明显的损伤累积效应，即随打击次数的增加而加重，3MCC组的损伤变化最为明显。（2）随着伤后时间推移，至伤后第6个月，3MCC组CTE大鼠的上述系列生物学变化已明显出现了与CTE流行病学研究一致的临床特征，直至伤后12个月仍然没有完全恢复。（3）本研究在国内首次创建了CTE大鼠模型，且更符合临床脑震荡实际发生状态，而具有国际前沿性；首次发现CTE大鼠437个差异蛋白质、发现多领域行为学与分子影像学变化规律。其大脑皮质蛋白组学研究结果一经发表，就得到国际学术界的高等关注，并被美国化学学会“ACS新闻周刊”给以专题评论报道。上述研究成果，为我国深入开展大鼠CTE病理发展机制、临床诊断标记分子筛查、药物干预效果评价等研究奠定了坚实基础。