Bambakidis Ted, Dekker Simone E, Sillesen Martin, Liu Baoling, Johnson Craig N, Jin Guang, de Vries Helga E, Li Yongqing, Alam Hasan B
1 Department of Surgery, University of Michigan Hospital , Ann Arbor, Michigan.
2 Department of Anesthesiology, Institute for Cardiovascular Research, VU University Medical Center , Amsterdam, the Netherlands .
J Neurotrauma. 2016 Aug 15;33(16):1514-21. doi: 10.1089/neu.2015.4163. Epub 2016 Apr 8.
Traumatic brain injury and hemorrhagic shock (TBI+HS) elicit a complex inflammatory response that contributes to secondary brain injury. There is currently no proven pharmacologic treatment for TBI+HS, but modulation of the epigenome has been shown to be a promising strategy. The aim of this study was to investigate whether valproic acid (VPA), a histone deacetylase inhibitor, modulates the expression of cerebral inflammatory gene profiles in a large animal model of TBI+HS. Ten Yorkshire swine were subjected to computer-controlled TBI+HS (40% blood volume). After 2 h of shock, animals were resuscitated with Hextend (HEX) or HEX+VPA (300 mg/kg, n = 5/group). Six hours after resuscitation, brains were harvested, RNA was isolated, and gene expression profiles were measured using a porcine microarray. Ingenuity Pathway Analysis® (IPA), gene ontology (GO), Parametric Gene Set Enrichment Analysis (PGSEA), and DAVID (Database for Annotation, Visualization, and Integrated Discovery) were used for pathway analysis. Key microarray findings were verified using real-time polymerase chain reaction (PCR). IPA analysis revealed that VPA significantly down-regulated the complement system (p < 0.001), natural killer cell communication (p < 0.001), and dendritic cell maturation (p < 0.001). DAVID analysis indicated that a cluster of inflammatory pathways held the highest rank and gene enrichment score. Real-time PCR data confirmed that VPA significantly down-expressed genes that ultimately regulate nuclear factor-kB (NF-kB)-mediated production of cytokines, such as TYROBP, TREM2, CCR1, and IL-1β. This high-throughput analysis of cerebral gene expression shows that addition of VPA to the resuscitation protocol significantly modulates the expression of inflammatory pathways in a clinically realistic model of TBI+HS.
创伤性脑损伤和失血性休克（TBI+HS）引发复杂的炎症反应，这会导致继发性脑损伤。目前尚无经证实的针对TBI+HS的药物治疗方法，但表观基因组的调控已被证明是一种有前景的策略。本研究的目的是调查组蛋白去乙酰化酶抑制剂丙戊酸（VPA）是否能在TBI+HS的大型动物模型中调节脑内炎症基因谱的表达。十只约克夏猪接受计算机控制的TBI+HS（失血40%血容量）。休克2小时后，动物用贺斯（HEX）或HEX+VPA（300mg/kg，每组n=5）进行复苏。复苏6小时后，采集大脑，分离RNA，并使用猪微阵列测量基因表达谱。使用通路分析软件（IPA）、基因本体论（GO）、参数化基因集富集分析（PGSEA）和DAVID（注释、可视化和综合发现数据库）进行通路分析。关键微阵列结果使用实时聚合酶链反应（PCR）进行验证。IPA分析显示，VPA显著下调补体系统（p<0.001）、自然杀伤细胞通讯（p<0.001）和树突状细胞成熟（p<0.001）。DAVID分析表明，一组炎症通路的排名和基因富集分数最高。实时PCR数据证实，VPA显著下调最终调节核因子-κB（NF-κB）介导的细胞因子产生的基因，如TYROBP、TREM2、CCR1和IL-1β。这种对脑基因表达的高通量分析表明，在复苏方案中添加VPA可在临床上逼真的TBI+HS模型中显著调节炎症通路的表达。
