Bailey Zachary S, Grinter Michael B, VandeVord Pamela J
Department of Biomedical Engineering and Mechanics, Virginia Tech Blacksburg, VA, USA.
Department of Biomedical Engineering and Mechanics, Virginia TechBlacksburg, VA, USA; Salem Veterans Affairs Medical CenterSalem, VA, USA.
Front Mol Neurosci. 2016 Aug 8;9:64. doi: 10.3389/fnmol.2016.00064. eCollection 2016.
Blast induced neurotrauma (BINT) is a prevalent injury within military and civilian populations. The injury is characterized by persistent inflammation at the cellular level which manifests as a multitude of cognitive and functional impairments. Epigenetic regulation of transcription offers an important control mechanism for gene expression and cellular function which may underlie chronic inflammation and result in neurodegeneration. We hypothesize that altered histone acetylation patterns may be involved in blast induced inflammation and the chronic activation of glial cells. This study aimed to elucidate changes to histone acetylation occurring following injury and the roles these changes may have within the pathology. Sprague Dawley rats were subjected to either a 10 or 17 psi blast overpressure within an Advanced Blast Simulator (ABS). Sham animals underwent the same procedures without blast exposure. Memory impairments were measured using the Novel Object Recognition (NOR) test at 2 and 7 days post-injury. Tissues were collected at 7 days for Western blot and immunohistochemistry (IHC) analysis. Sham animals showed intact memory at each time point. The novel object discrimination decreased significantly between two and 7 days for each injury group (p < 0.05). This is indicative of the onset of memory impairment. Western blot analysis showed glial fibrillary acidic protein (GFAP), a known marker of activated astrocytes, was elevated in the prefrontal cortex (PFC) following blast exposure for both injury groups. Analysis of histone protein extract showed no changes in the level of any total histone proteins within the PFC. However, acetylation levels of histone H2b, H3, and H4 were decreased in both groups (p < 0.05). Co-localization immunofluorescence was used to further investigate any potential correlation between decreased histone acetylation and astrocyte activation. These experiments showed a similar decrease in H3 acetylation in astrocytes exposed to a 17 psi blast but not a 10 psi blast. Further investigation of gene expression by polymerase chain reaction (PCR) array, showed dysregulation of several cytokine and cytokine receptors that are involved in neuroinflammatory processes. We have shown aberrant histone acetylation patterns involved in blast induced astrogliosis and cognitive impairments. Further understanding of their role in the injury progression may lead to novel therapeutic targets.
爆炸所致神经创伤(BINT)在军人和平民群体中都是一种常见损伤。这种损伤的特征是细胞水平上的持续炎症,表现为多种认知和功能障碍。转录的表观遗传调控为基因表达和细胞功能提供了一种重要的控制机制,这可能是慢性炎症的基础并导致神经退行性变。我们推测组蛋白乙酰化模式的改变可能与爆炸诱导的炎症以及胶质细胞的慢性激活有关。本研究旨在阐明损伤后组蛋白乙酰化的变化以及这些变化在病理过程中可能发挥的作用。将Sprague Dawley大鼠置于先进爆炸模拟器(ABS)中,使其承受10或17磅力/平方英寸的爆炸超压。假手术动物接受相同程序但不进行爆炸暴露。在损伤后2天和7天使用新颖物体识别(NOR)测试来测量记忆损伤。在7天时收集组织用于蛋白质免疫印迹和免疫组织化学(IHC)分析。假手术动物在每个时间点都显示出完整的记忆。每个损伤组在2天和7天之间新颖物体辨别能力显著下降(p < 0.05)。这表明记忆损伤的开始。蛋白质免疫印迹分析显示,对于两个损伤组,在爆炸暴露后,前额叶皮质(PFC)中胶质纤维酸性蛋白(GFAP,一种已知的活化星形胶质细胞标志物)升高。组蛋白提取物分析显示PFC内任何总组蛋白的水平没有变化。然而,两组中组蛋白H2b、H3和H4的乙酰化水平均降低(p < 0.05)。共定位免疫荧光用于进一步研究组蛋白乙酰化降低与星形胶质细胞激活之间的任何潜在相关性。这些实验显示,暴露于17磅力/平方英寸爆炸的星形胶质细胞中H3乙酰化有类似程度的降低,但暴露于10磅力/平方英寸爆炸的星形胶质细胞中没有。通过聚合酶链反应(PCR)阵列对基因表达的进一步研究表明,参与神经炎症过程的几种细胞因子和细胞因子受体存在失调。我们已经证明爆炸诱导的星形胶质细胞增生和认知障碍涉及异常的组蛋白乙酰化模式。进一步了解它们在损伤进展中的作用可能会导致新的治疗靶点。