C1型尼曼-匹克症小鼠脊髓胶质细胞的活性变化

杨恩慧 乔梁 林俊堂* 闫欣*

doi:10.16098/j.issn.0529-1356.2017.03.003

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解剖学报 ›› 2017, Vol. 48 ›› Issue (3) : 260-265. DOI: 10.16098/j.issn.0529-1356.2017.03.003

神经生物学

C1型尼曼-匹克症小鼠脊髓胶质细胞的活性变化

杨恩慧^{1, 2}乔梁^{1, 2}林俊堂^{1, 2*}闫欣^{1, 2*}

作者信息 +

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

YANG En-hui^{1, 2} QIAO Liang^{1, 2} LIN Jun-tang ^{1, 2*} YAN Xin ^{1, 2*}

Author information +

文章历史 +

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Npc1^-/-（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。

Abstract

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 Npc1^-/- spinal cord. Methods Npc1^+/- mice bred to generate Npc1^-/- mice （n=3）and wild type mice (Npc1^+/+ )（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 Npc1-/- and Npc1+/+ mice at the postnatal day 35. Double Immunofluorescent staining using GFAP and F4/80 with interleukin-1β(IL-1β) investigated the distribution of IL-1β in the Npc1-/- spinal cord. IL-1β, SMI31 and phos-tau were detected by Western blotting. Results GFAP and F4/80 immunofluorescent staining indicated a significantly increased glial activation (P<0.05) in both dorsal and ventral horn of Npc1^-/- spinal cord, which was associated with enhanced IL-1β 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.

导出引用

杨恩慧乔梁林俊堂闫欣. C1型尼曼-匹克症小鼠脊髓胶质细胞的活性变化[J]. 解剖学报. 2017, 48(3): 260-265 https://doi.org/10.16098/j.issn.0529-1356.2017.03.003

YANG En-hui QIAO Liang LIN Jun-tang YAN Xin. Activation of glial cells in the spinal cord of Niemann-Pick disease type C1 mice[J]. Acta Anatomica Sinica. 2017, 48(3): 260-265 https://doi.org/10.16098/j.issn.0529-1356.2017.03.003

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基金

C1型尼曼-匹克氏症轴突发育异常的病理机制;C1型尼曼-匹克氏症的神经发育基础研究;miR-205在C1型尼曼-匹克病小鼠髓鞘形成中的作用机制研究;microRNA-205调控tau蛋白磷酸化在C1型尼曼-匹克病中的作用机制及潜在治疗研究;钙粘蛋白在神经发育中的作用及对干细胞诱导分化的影响;鸡胚模型中研究ADAM17超表达引起微血管增生及促进人神经瘤发展的机制