CENTRAL NERVOUS SYSTEM DYSFUNCTION IN SHOCK AND TRAUMA

描述（改编自摘要）：使用两者的已建立模型 失代偿性出血性低血压（HH）、创伤性脑损伤（TBI）和 联合出血/脑损伤范例（HH-TBI），本申请提出 (1)为了表征和阐明细胞死亡/存活基因的作用， 细胞因子和DNA损伤检测和修复机制的改变， 介导出血后中枢神经系统（CNS）功能障碍， 机械损伤和（2）评价新的药物治疗策略 针对这些途径治疗休克和创伤。中央 假设是休克后脑中细胞死亡和功能障碍 创伤是由于死亡诱导基因（如Bax， 半胱氨酸蛋白酶-3，白细胞介素，TNF-α）和下调神经保护性 基因（如Bcl-2，Bcl-xL），以及大脑中的分子变化 单次损伤后观察到的结果将不同于组合损伤后观察到的结果 休克和脑外伤Specific Aim 1将使用原位杂交/ 免疫组织化学来评估HH或TBI如何导致在细胞中的改变。 大脑中细胞死亡/存活基因的平衡（比率）， 这些损伤后的凋亡细胞死亡。转基因的反应 动物，通过基因工程过度表达抗凋亡基因bcl-2， 和药理学抑制促凋亡因子的功效。 将评估蛋白胱天蛋白酶-3。特异性目的2评价基因表达 HH、TBI或HH-TBI后大鼠脑中炎性细胞因子的变化， 表征炎症级联在介导CNS功能障碍中的作用。 转基因小鼠，经基因工程改造以缺乏TNF-α表达 (TNF-/-小鼠），将用于验证假设;和治疗性小鼠， 重组人白细胞介素-18受体拮抗剂抑制 将评价细胞因子功能。具体目标3将评估效果 HH、TBI或HH-TBI对内源性DNA损伤检测和修复机制的影响 （激活PARP）。PARP敲除小鼠（PARP KO）将用于验证 假设这些损伤改变了DNA修复机制， 细胞死亡和功能障碍。PARP的药理学抑制作用也将是 评估。Specific Aim 4使用单细胞aRNA扩增技术， 从显示DNA的神经元中获得多个基因的表达谱 在HH、TBI或HH-TBI后大鼠脑中的碎片，以建立 协调的基因组变化可能在细胞死亡和功能障碍中发挥作用。 总之，这些研究将大大提高对 CNS对出血性休克、TBI和联合损伤的分子反应， 将导致更有效的治疗方法的发展， 治疗休克和创伤。9;

DESCRIPTION (adapted from the abstract):	Using established models of both uncompensated hemorrhagic hypotension (HH), traumatic brain injury (TBI) and a combined hemorrhage/brain injury paradigm (HH-TBI), this application proposes (1) to characterize and elucidate the role of cell death/survival genes, cytokines and alterations in DNA damage detection and repair mechanisms in mediating central nervous system (CNS) dysfunction following hemorrhage and mechanical injury and (2) evaluate novel pharmacotherapeutic strategies targeted at these pathways in the treatment of shock and trauma. The central hypothesis is that cellular death and dysfunction in the brain after shock or trauma is due to an up regulation of death-inducing genes (such as bax, capase-3, interleukins, TNF-alpha) and a downregulation of neuroprotective genes (such as Bcl-2, Bcl-xL), and that molecular changes in the brain following the single insults will differ from those observed following combined shock and brain trauma. Specific Aim 1 will use in situ hybridization/ immunohistochemistry to evaluate how HH or TBI results in an alteration in the balance (ratio) of cell death/survival genes in the brain which contribute to apoptotic cell death following these insults. The response of transgenic animals, genetically engineered to over express the anti-apoptotic gene bcl-2, to HH or TBI and the efficacy of pharmacologic inhibition of the pro-apoptotic protein caspase-3 will be evaluated. Specific Aim 2 evaluates gene expression of inflammatory cytokines in the rat brain following HH, TBI or HH-TBI to characterize the role of inflammatory cascades in mediating CNS dysfunction. Transgenic mice, genetically engineered to be deficient in expression of TNF-a (TNF-/- mice), will be used to validate the hypothesis; and the therapeutic efficacy of recombinant human interleukin-18 receptor antagonist to inhibit cytokine function will be evaluated. Specific Aim 3 will evaluate the effects of HH, TBI or HH-TBI on endogenous DNA damage detection and repair mechanisms (activation of PARP). PARP knockout mice (PARP KO) will be used to validate out hypothesis that these insults alter DNA repair mechanisms which contributes to cell death and dysfunction. Pharmacologic inhibition of PARP will also be evaluated. Specific Aim 4 use single-cell aRNA amplification techniques to obtain expression profiles of multiple genes from neurons exhibiting DNA fragmentation in rat brain following HH, TBI or HH-TBI to establish the coordinated genomic changes that may play a role in cell death and dysfunction. Taken together, these studies will significantly enhance understanding of the molecular response in the CNS to hemorrhage shock, TBI, and combined injury and will result in the development of more effective therapeutic approaches to the treatment of shock and trauma.