CENTRAL NERVOUS SYSTEM DYSFUNCTION IN SHOCK AND TRAUMA

休克和创伤中的中枢神经系统功能障碍

基本信息

批准号：
6476457
负责人：
TRACY K. MCINTOSH
金额：
$ 23.96万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
1988
资助国家：
美国
起止时间：
1988-06-01 至 2003-11-30
项目状态：
已结题

项目摘要

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.	

描述（改编自摘要）：使用两者的既定模型失代偿性失血性低血压（HH）、创伤性脑损伤（TBI）和联合出血/脑损伤范例（HH-TBI），该申请提出 (1) 表征和阐明细胞死亡/存活基因的作用，细胞因子以及 DNA 损伤检测和修复机制的改变介导出血后的中枢神经系统（CNS）功能障碍机械损伤和（2）评估新的药物治疗策略针对休克和创伤治疗中的这些途径。中央假设是休克或休克后大脑中的细胞死亡和功能障碍创伤是由于死亡诱导基因（例如bax、 capase-3、白细胞介素、TNF-α）和神经保护作用的下调基因（例如 Bcl-2、Bcl-xL）以及大脑中的分子变化单一侮辱后观察到的结果将不同于组合侮辱后观察到的结果休克和脑外伤。具体目标1将使用原位杂交/ 免疫组织化学来评估 HH 或 TBI 如何导致大脑中细胞死亡/存活基因的平衡（比率）有助于这些损伤后细胞凋亡。转基因的反应经过基因工程改造的动物过度表达抗凋亡基因 bcl-2， HH 或 TBI 以及药物抑制促凋亡细胞的功效将评估蛋白质 caspase-3。具体目标 2 评估基因表达 HH、TBI 或 HH-TBI 后大鼠脑中炎症细胞因子的变化描述炎症级联在介导中枢神经系统功能障碍中的作用。转基因小鼠，经过基因工程改造，缺乏 TNF-a 的表达（TNF-/- 小鼠），将用于验证假设；和治疗的重组人白细胞介素18受体拮抗剂的抑制作用将评估细胞因子功能。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，从而导致细胞死亡和功能障碍。 PARP 的药理学抑制也将评价。具体目标 4 使用单细胞 aRNA 扩增技术从表现出 DNA 的神经元中获取多个基因的表达谱 HH、TBI 或 HH-TBI 后大鼠大脑中的碎片化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。总而言之，这些研究将显着增强对中枢神经系统对失血性休克、创伤性脑损伤和复合损伤的分子反应将导致开发出更有效的治疗方法休克和创伤的治疗。