Translational Gerontology Branch, Intramural Research Program, National Institute on Aging, NIH, Baltimore, MD 21224, USA.
Department of Anatomy and Anthropology, Sackler School of Medicine, Tel-Aviv University, Tel Aviv 69978, Israel.
Neurobiol Dis. 2019 Oct;130:104528. doi: 10.1016/j.nbd.2019.104528. Epub 2019 Jul 8.
Mild traumatic brain injury (mTBI) is a risk factor for neurodegenerative disorders, such as Alzheimer's disease (AD) and Parkinson's disease (PD). TBI-derived neuropathologies are promoted by inflammatory processes: chronic microgliosis and release of pro-inflammatory cytokines that further promote neuronal dysfunction and loss. Herein, we evaluated the effect on pre-programmed cell death/neuroinflammation/synaptic integrity and function of (-)-Phenserine tartrate (Phen), an agent originally developed for AD. This was studied at two clinically translatable doses (2.5 and 5.0 mg/kg, BID), in a weight drop (concussive) mTBI model in wild type (WT) and AD APP/PSEN1 transgenic mice. Phen mitigated mTBI-induced cognitive impairment, assessed by Novel Object Recognition and Y-maze behavioral paradigms, in WT mice. Phen fully abated mTBI-induced neurodegeneration, evaluated by counting Fluoro-Jade C-positive (FJC+) cells, in hippocampus and cortex of WT mice. In APP/PSEN1 mice, degenerating cell counts were consistently greater across all experimental groups vs. WT mice. mTBI elevated FJC+ cell counts vs. the APP/PSEN1 control (sham) group, and Phen similarly mitigated this. Anti-inflammatory effects on microglial activation (IBA1-immunoreactivity (IR)) and the pro-inflammatory cytokine TNF-α were evaluated. mTBI increased IBA1-IR and TNF-α/IBA1 colocalization vs. sham, both in WT and APP/PSEN1 mice. Phen decreased IBA1-IR throughout hippocampi and cortices of WT mice, and in cortices of AD mice. Phen, likewise, reduced levels of IBA1/TNF-α-IR colocalization volume across all areas in WT animals, with a similar trend in APP/PSEN1 mice. Actions on astrocyte activation by mTBI were followed by evaluating GFAP, and were similarly mitigated by Phen. Synaptic density was evaluated by quantifying PSD-95+ dendritic spines and Synaptophysin (Syn)-IR. Both were significantly reduced in mTBI vs. sham in both WT and APP/PSEN1 mice. Phen fully reversed the PSD-95+ spine loss in WT and Syn-IR decrease in both WT and APP/PSEN1 mice. To associate immunohistochemical changes in synaptic markers with function, hippocampal long term potentiation (LTP) was induced in WT mice. LTP was impaired by mTBI, and this impairment was mitigated by Phen. In synopsis, clinically translatable doses of Phen ameliorated mTBI-mediated pre-programmed cell death/neuroinflammation/synaptic dysfunction in WT mice, consistent with fully mitigating mTBI-induced cognitive impairments. Phen additionally demonstrated positive actions in the more pathologic brain microenvironment of AD mice, further supporting consideration of its repurposing as a treatment for mTBI.
轻度创伤性脑损伤 (mTBI) 是神经退行性疾病(如阿尔茨海默病 [AD] 和帕金森病 [PD])的危险因素。源自创伤性脑损伤的神经病理学由炎症过程促进:慢性小胶质细胞增生和促炎细胞因子的释放,进一步促进神经元功能障碍和丧失。在此,我们评估了 (-)-苯并恶嗪磷酸盐(Phen)对预先编程的细胞死亡/神经炎症/突触完整性和功能的影响,Phen 最初是为 AD 开发的。这是在两种具有临床转化潜力的剂量(2.5 和 5.0 mg/kg,BID)下,在野生型(WT)和 AD APP/PSEN1 转基因小鼠的重物跌落(震荡性)mTBI 模型中进行的。Phen 减轻了 WT 小鼠中由新奇物体识别和 Y 迷宫行为范式评估的 mTBI 引起的认知障碍。Phen 完全消除了 WT 小鼠中海马和皮质中由 Fluoro-Jade C 阳性(FJC+)细胞计数评估的 mTBI 诱导的神经退行性变。在 APP/PSEN1 小鼠中,所有实验组的变性细胞计数均明显高于 WT 小鼠。与 APP/PSEN1 对照(假手术)组相比,mTBI 增加了 FJC+细胞计数,而 Phen 则同样减轻了这一计数。评估了对小胶质细胞激活(IBA1-免疫反应性(IR))和促炎细胞因子 TNF-α 的抗炎作用。与假手术组相比,mTBI 增加了 WT 和 APP/PSEN1 小鼠的 IBA1-IR 和 TNF-α/IBA1 共定位。Phen 降低了 WT 小鼠海马和皮质以及 AD 小鼠皮质中 IBA1-IR 的水平。Phen 同样降低了 WT 动物所有区域中 IBA1/TNF-α-IR 共定位体积,在 APP/PSEN1 小鼠中也有类似的趋势。通过评估 GFAP 来跟踪 mTBI 对星形胶质细胞激活的作用,并通过 Phen 同样减轻了这种作用。通过量化 PSD-95+树突棘和突触素(Syn)-IR 来评估突触密度。与假手术组相比,mTBI 使 WT 和 APP/PSEN1 小鼠中的这两种标志物均显著减少。Phen 完全逆转了 WT 小鼠中的 PSD-95+棘突丢失和 WT 和 APP/PSEN1 小鼠中的 Syn-IR 减少。为了将突触标志物的免疫组织化学变化与功能相关联,在 WT 小鼠中诱导了海马长时程增强(LTP)。mTBI 损害了 LTP,而 Phen 减轻了这种损害。总之,临床转化剂量的 Phen 改善了 WT 小鼠中由 mTBI 介导的预先编程的细胞死亡/神经炎症/突触功能障碍,与完全减轻 mTBI 引起的认知障碍一致。Phen 还在 AD 小鼠更具病理性的大脑微环境中表现出积极作用,进一步支持将其重新用于治疗 mTBI 的考虑。