MACROPHAGES/MICROGLIA AND CSPGS IN SPINAL CORD INJURY

脊髓损伤中的巨噬细胞/小胶质细胞和 CSPGS

• 批准号: 6187622
• 负责人: LEONARD L WERNER-JONES
• 金额: $ 3.24万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6187622
• 关键词: axon; chondroitin sulfates; extracellular matrix; fibroblasts; genetically modified animals; growth factor; immunocytochemistry; in situ hybridization; interleukin 10; laboratory rat; macrophage; microglia; nervous system regeneration; neuronal guidance; polymerase chain reaction; proteoglycan; spinal cord injury; tissue /cell culture

Over 250,000 Americans are severely and permanently disabled due to acute spinal cord injuries (SCI). There currently is no therapy for promoting recovery of neurological function in chronic stages of SCI. The proposed study will test the central hypothesis that macrophages/microglia express matrix molecules that inhibit axonal outgrowth after SCI, and that the blockade of these putative inhibitory molecules will augment axonal regeneration. To challenge this hypothesis, the following Specific Aims will be addressed: 1) Determine the cellular source of CSPG proteoglycan expression using in situ hybridization and immunolabelling, and whether the inhibition of proteoglycan synthesis by application of the proteoglycan inhibitor beta-D- xyloside enhances axonal outgrowth after SCI; 2) Determine using semi-quantitative PCR whether macrophages treated with growth factors and cytokines modulate their expression of CSPGs in vitro; and 3) Determine whether grafts of fibroblasts genetically modified to express and secrete augmented amounts of inhibitory cytokines (e.g., interleukin 10 or other substances identified in Aims 2) will reduce the deposition of ECM molecules and promote axonal repair after SCI. Here, transgenic cellular delivery, immunolabeling, in situ hybridization and neuroanatomical tracing will be used.

超过25万美国人因急性脊髓损伤（SCI）而严重和永久残疾。目前还没有一种治疗方法可以促进慢性脊髓损伤患者神经功能的恢复。该研究将验证一个中心假设，即巨噬细胞/小胶质细胞表达抑制脊髓损伤后轴突生长的基质分子，并且阻断这些假定的抑制分子将增强轴突再生。为了挑战这一假设，将解决以下具体目标：1)利用原位杂交和免疫标记确定CSPG蛋白多糖表达的细胞来源，以及应用蛋白多糖抑制剂β - d -木糖苷抑制蛋白多糖合成是否能促进脊髓损伤后轴突的生长；2)采用半定量PCR检测生长因子和细胞因子对巨噬细胞体外CSPGs表达的调节作用；3)确定经基因修饰的成纤维细胞移植物是否能表达和分泌更多的抑制性细胞因子（如白细胞介素10或Aims 2中发现的其他物质），从而减少脊髓损伤后ECM分子的沉积，促进轴突修复。在这里，将使用转基因细胞递送、免疫标记、原位杂交和神经解剖示踪。