甲基强的松龙对体大鼠脊髓损伤后内源性神经肝细胞增殖与迁移的影响.pdf

目 录 1 甲基强的松龙对成体大鼠脊髓损伤后内源性神经干细胞增殖 与迁移的影响„„„„„„„„„„„„„„„„„„„1 1.1 中文摘要„„„„„„„„„„„„„„„„„„„„„1 1.2 英文摘要„„„„„„„„„„„„„„„„„„„„„3 1.3 前言„„„„„„„„„„„„„„„„„„„„„„„5 1.4 材料与方法„„„„„„„„„„„„„„„„„„„„7 1.5 结果„„„„„„„„„„„„„„„„„„„„„„„12 1.6 讨论„„„„„„„„„„„„„„„„„„„„„„„24 1.7 结论„„„„„„„„„„„„„„„„„„„„„„„30 1.8 参考文献„„„„„„„„„„„„„„„„„„„„„31 1.9 英汉缩略词对照表„„„„„„„„„„„„„„„„„35 2 致谢„„„„„„„„„„„„„„„„„„„„„„„36 3 脊髓损伤后炎症反应与脊髓内源性神经干细胞（综述） „„„„„„„„„„„„„„„„„„„„„„„„„39 甲基强的松龙对成体大鼠脊髓损伤后内源性神经 干细胞增殖与迁移的影响 摘 要 目的：目前认为在大脑和脊髓中存在内源性神经干细胞 （endogenous neural stem cells,ENSCs ）,且在脊髓损伤后发生了增殖， 分化与迁移。甲强龙曾一度作为急性脊髓损伤的首选药用于临床，本 实验着眼于脊髓损伤后内源性神经干细胞的生物学行为探讨甲基强 的松龙（MP ）对成体大鼠脊髓损伤后内源性神经干细胞的增殖与迁 移的影响。方法：①实验材料：将 60 只成年雌性 SD 大鼠随机分为 甲强龙组（30 只），生理盐水组（30 只）。②实验方法：所有动物均 采用 Allen 法建立 T10 平面完全性截瘫模型，MP 以 0.9%NS 配置成 浓度为 1mg/ml 的溶液，MP 组大鼠于 1 小时内经尾静脉缓慢推注 MP （30mg/kg ）,随后按照 5.4mg/(kg..h)算出 23 小时总量，分 3 次（8h/ 次）缓慢尾静脉注射。生理盐水组以同样方法注射等量的生理盐水。 Brdu （5-溴 2-脱氧尿嘧啶核苷）在脊髓损伤后 2h 内按照 50mg/kg 的 剂量腹腔注射连续 10 天。③实验评估：在模型制作后 14d 、21d 、28d 每组大鼠各取 10 只分别取出损伤脊髓节段，采用免疫组织化学方法 检测Brdu 标记的内源性神经干细胞的增殖及迁移情况;BBB 运动等级 评分检测两组大鼠下肢运动功能。结果：①两组动物的脊髓切片均可 见大量的 Brdu （5-溴 2-脱氧尿嘧啶核苷）阳性细胞存在，这些细胞 主要分布于脊髓损伤区域周围。②甲强龙组在各个时间点脊髓切片中 的Brdu （5-溴 2-脱氧尿嘧啶核苷）阳性细胞较生理盐水组明显减少； 以脊髓损伤中心作切片发现生理盐水组 Brdu 阳性细胞数目在各个时 间点明显较多。③两组大鼠的脊髓切片中出现大量 Brdu （5-溴 2-脱 氧尿嘧啶核苷）阳性细胞是在脊髓损伤后 14d 左右，21d,28d 后逐渐 减少。④两组大鼠的 BBB 评分在各个时间点无明显差异，且随着时 间的推移无明显变化。结论：脊髓损伤后将发生内源性神经干细胞的 增殖，迁移且在损伤后 14 天达到高峰：甲强龙可抑制脊髓损伤后内 源性神经干细胞的增殖与迁移。 关键词 甲强龙 内源性神经干细胞 脊髓损伤 增殖 迁移 大鼠 MP Affect On EndoGenous Neural Stem Cell Proliferation And Migration After The Adult Rat Spinal Cord Injury Abstract:Objective : That the existence of endogenous neural stem cells in the brain and spinal cord, And after spinal