C1型尼曼-匹克症小鼠脊髓胶质细胞的活性变化  

Activation of glial cells in the spinal cord of Niemann-Pick disease type C1 mice

作　　者：杨恩慧[1,2] 乔梁[1,2] 林俊堂[1,2] 闫欣[1,2] YANG En-hui QIAO Liang LIN Jun-tang YAN Xin(College of Life Science and Technology, Xinxiang Medical University, He＇ nan Xinxiang 453003, China He＇ nan Key Laboratory of Medical Tissue Regeneration, He＇ nan Xinxiang 453003, China)

机构地区：[1]新乡医学院生命科学技术学院,河南新乡453003 [2]河南省医用组织再生重点实验室,河南新乡453003

出　　处：《解剖学报》2017年第3期260-265,共6页Acta Anatomica Sinica

基　　金：国家自然科学基金(81400936;U1304808;81600987);河南省高等学校重点科研项目(16A180014);河南省高校科技创新人才支持计划(14HASTIT032);河南省科技发展计划(142300410192);新乡医学院研究生科研创新支持计划(YJSCX201516Y)

摘　　要：目的通过观察C1型尼曼-匹克症小鼠不同脊髓节段星形胶质细胞和小胶质细胞的活性变化,探讨Npc1基因突变对脊髓发育的影响。方法 Npc1^(+/-)小鼠交配繁殖产生Npc^(1-/-)(n=3)和Npc1^(+/+)小鼠(n=3),PCR检测新生小鼠的基因型;选取35日龄的Npc1^(-/-)和Npc1^(+/+)小鼠,采用免疫荧光方法观察对比脊髓不同节段(颈、胸、腰、骶)星形胶质细胞和小胶质细胞的活性变化。采用免疫双染色检测胶质细胞中炎性因子的表达情况,采用免疫印迹方法检测白细胞介素-1β(IL-1β)、SMI31和磷酸化的tau蛋白表达情况。结果在35日龄Npc1^(-/-)小鼠脊髓的各个节段,其背角和腹角的星形胶质细胞和小胶质细胞的活性均明显增强(P<0.05),并伴随细胞炎性因子IL-1β表达量的显著增加;同时,脊髓神经丝蛋白和骨架蛋白tau蛋白发生超磷酸化。结论 Npc1基因突变引起脊髓神经胶质细胞发生病理性变化,可能是脊髓神经元病理性损伤的重要原因。Objective To explore the impact of Niemann-pick disease type C1 （NPC1） on the developing spinal cord by observing the activation of astrocytes and microglia in different segments of Npcl-/- spinal cord. Methods Npcl ＋/ mice bred to generate Npcl -/- mice （n=3） and wild type mice （Npcl ＋/＋）（n=3）, and the mice genotypes were detected by PCR. Immunofluorescent staining was performed on different levels of spinal cord （cervical, thoracic, lumbar, sacrum） and the activations of astrocytes and microglia were compared between Npcl -/- and Npcl ＋/＋ mice at the postnatal day 35. Double Immunofluorescent staining using GFAP and F4/80 with interleukin-113 （IL-113） investigated the distribution of IL-113 in the Npcl -/- spinal cord. IL-113, SMI31 and phos-tau were detected by Western blotting. Results GFAP and F4/80 immunofluorescent staining indicated a significantly increased glial activation （ P 〈 O. 05 ） in both dorsal and ventral horn of Npcl -/- spinal cord, which was associated with enhanced IL-1 13 expression in the glial cells. Western blotting indicated that an up-regulation of phosphorylated neurofilaments and tau protein resulted in axon accumulation in NPC1 -/- spinal cord. Conclusion Our data show pathological changes of glial cells in the NPC1 -/- spinal cord, which is the possible reason of neuronal defects in the NPC1 -/- mice.

关 键 词：C1型尼曼-匹克症 脊髓 星形胶质细胞 小胶质细胞 免疫印迹法 小鼠 

分 类 号：Q954.48[生物学—动物学]