CENTRAL NERVOUS SYSTEM DYSFUNCTION IN SHOCK AND TRAUMA

描述(改编自摘要)：使用两者的既定模型 失代偿性出血性低血压(HH)、创伤性脑损伤(TBI)和 联合出血/脑损伤范式(HH-TBI)，该应用程序提出 (1)鉴定和阐明细胞死亡/存活基因的作用， 细胞因子与DNA损伤检测和修复机制的改变 失血性休克后中枢神经系统功能障碍的调节 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。中环 假说是休克或休克后脑细胞死亡和功能障碍 创伤是由于死亡诱导基因(如bax， CAPase-3、IL-6、TNF-α)和神经保护的下调 基因(如Bcl2、Bclxl)，以及大脑中的分子变化 单次侮辱之后的侮辱将与随后观察到的联合侮辱不同 休克和脑损伤。特定目标1将使用原位杂交/ 免疫组织化学方法评估HH或TBI如何导致脑组织的改变 大脑中细胞死亡/存活基因的平衡(比率)，这有助于 在这些侮辱之后，细胞的凋亡死亡。转基因生物的反应 通过基因工程过量表达抗细胞凋亡基因bcl2的动物， 对HH或TBI的影响及药物抑制促凋亡作用 将对caspase-3蛋白进行评估。特定目标2评估基因表达 HH、TBI或HH-TBI后大鼠脑内炎性细胞因子的变化 描述炎症级联在调节中枢神经系统功能障碍中的作用。 转基因小鼠，转基因小鼠缺乏肿瘤坏死因子-a的表达 (肿瘤坏死因子-/-小鼠)，将被用来验证这一假说； 重组人白介素18受体拮抗剂的抑制作用 将对细胞因子功能进行评估。具体目标3将评估效果 HH、TBI和HH-TBI对内源性DNA损伤检测和修复机制的影响 (激活PARP)。PARP基因敲除小鼠(PARP KO)将用于验证 假设这些侮辱会改变DNA修复机制，从而导致 细胞死亡和功能障碍。PARP的药理抑制也将是 已评估。特定目标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 大鼠HH、TBI或HH-TBI后脑组织的碎片化 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。 综上所述，这些研究将大大加强对 失血性休克、脑外伤及复合伤后中枢神经系统的分子反应 将导致开发出更有效的治疗方法 休克和创伤的治疗。

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.