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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 investigate the effect of Npc1 gene mutation on the development of spinal cord by observing the changes of the activity of astrocytes and microglia in different spinal cord segments of type-1 Niemann-Pick disease. Methods Npc 1 ~ (+/-) mice were intercropped to produce Npc ~ (1 - / -) (n = 3) and Npc1 ~ (+ ; 35-day-old Npc1 ~ (- / -) and Npc1 ~ (+ / +) mice were selected and immunofluorescence staining was used to observe the changes of astrocytes in different segments of the spinal cord (neck, thoracolumbar, Glial cell activity changes. Immunofluorescence staining was used to detect the expression of inflammatory cytokines in glial cells. The expression of interleukin-1β (IL-1β), SMI31 and phosphorylated tau protein were detected by Western blotting. Results The activities of astrocytes and microglia in the dorsal horn and ventral horn of Npc1 ~ (- / -) mice at 35-day-old were significantly increased (P <0.05) The expression of IL-1β was significantly increased. Meanwhile, spinal neurofilament and skeletal protein tau hyperphosphorylated. Conclusion The mutation of Npc1 causes pathological changes of glial cells in spinal cord and may be an important reason of pathological injury of spinal neurons.