REGULATION OF INTERLEUKIN-6 IN THE AGING BRAIN

描述（申请人摘要）：炎症的表达和作用 大脑中的细胞因子分子需要严格的调节，因为它们会改变 认知和动机系统，如果过度表达，可能是神经毒性的。 本申请中提供的初步数据表明，培养的小胶质细胞 从老年小鼠的大脑自发分泌大量的 炎性细胞因子白细胞介素-6（IL-6）。因此，积极的 已经发现年龄与脑中IL-6浓度之间的相关性，即使在 健康的老鼠我们认为，IL-6基因的慢性过度表达， 老年人的大脑可能会建立一个允许发病的状态， 神经退行性疾病，并与厌食症和受损有因果关系 记忆和学习不幸的是，人们对衰老 影响IL-6基因在大脑中的表达，这是本提案的重点。 在这里，提出了几种方法来（1）定义IL-6在细胞中的表达， （2）确定年龄对转录的影响 调节脑和神经胶质细胞中IL-6基因表达的因子;（3） 探讨老年小鼠脑IL-6基因表达的调控能力 补充脱氢表雄酮（DHEA），和（4）描述的影响 DHEA对IL-6基因转录调控因子的影响。我们特别 将采用免疫组织化学和原位杂交来绘制IL-6 表达，双色流式细胞仪检测老年小鼠脑中的 从新生儿、成人、 和表达IL-6的老年小鼠。我们将确定IL-6的增加是否 在老年大脑中的产生是由于活性核因子-KB的增加或 重组信号序列结合蛋白JK的减少， 最近显示通过占据IL-6-KB抑制自发IL-6产生 IL-6启动子/增强子区域中的位点。此外，关键的可能性 调节IL-6基因转录因子的年龄相关变化 在脑中的表达是由于DHEA的下降，这随着年龄的增长而发生， 会得到考验这些新的研究将提供所需的信息， 了解，治疗或预防老年人的神经病理生理表现 年龄有强有力的初步数据支持每一个目标， 成功完成这些小鼠研究所需的所有技术 已经被开发出来了。

DESCRIPTION (applicant's abstract): The expression and action of inflammatory cytokine molecules in the brain requires tight regulation since they alter cognitive and motivational systems and if over-expressed can be neurotoxic. Preliminary data presented in this application indicate that microglia cultured from brain of aged mice spontaneously secrete copious amounts of the inflammatory cytokine, interleukin-6 (IL-6). Consequently, a positive correlation between age and IL-6 concentration in brain has been found, even in healthy mice. We believe that chronic over-expression of the IL-6 gene in the brain of the elderly may establish a state that is permissive to the onset of neurodegenerative disease and be causally related to anorexia and impaired memory and learning. Unfortunately, little to nothing is known about how aging affects IL-6 gene expression in the brain, which is the focus of this proposal. Here, several approaches are proposed to (1) define the expression of IL-6 in the brain of aged mice, (2) determine the effects of age on the transcription factors that regulate IL-6 gene expression in brain and in glial cells; (3) explore the ability to regulate IL-6 gene expression in the brain of aged mice with supplemental dehydroepiandrosterone (DHEA), and (4) describe the effects of DHEA on the transcription factors regulating the IL-6 gene. Specifically, we will employ immunohistochemistry and in situ hybridization to map IL-6 expression in aged mouse brain, and two-color flow cytometry to evaluate the proportion of astrocytes and microglia cultured from brain of neonate, adult, and aged mice expressing IL-6. We will determine if the increase in IL-6 production in aged brain is due to an increase in active Nuclear Factor-KB or a decrease in Recombinant Signal Sequence Binding-Protein JK, which has been recently shown to inhibit spontaneous IL-6 production by occupying the IL-6-KB site in the IL-6 promoter/enhancer region. Moreover, the critical possibility that the age-related change(s) in transcription factors regulating IL-6 gene expression in the brain is due to the decline in DHEA, which occurs with age, will be tested. These novel studies will provide needed information to better understand, treat, or prevent the neuropathophysiological manifestations of old age. There are strong preliminary data supporting each of the objectives and all of the techniques needed to successfully complete these studies in mice have been developed.