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目的研究Cuprizone(CPZ)诱导C57BL/6小鼠脱髓鞘对胼胝体少突胶质细胞变化情况,并探讨对小鼠运动能力的调控特点。方法以正常饲料喂食的6周龄雄性小鼠作为对照,实验组用含0.2%Cuprizone的混合饲料持续喂食6周龄雄性小鼠6周,诱导中枢神经系统脱髓鞘。用LFB组化和MBP免疫组化技术鉴定脑胼胝体区的脱髓鞘改变,确认模型的稳定可靠性。结合MBP和CC-1免疫荧光双标技术,观察脱髓鞘的胼胝体区少突胶质细胞变化情况。并通过体质量监测观察小鼠的整体功能状态变化,进一步采用行为学实验技术(旋转棒测试和旷场实验)观察对小鼠运动能力的调控效果和特点。结果与正常组相比,Cuprizone诱导脱髓鞘6周后,脑胼胝体区的髓鞘缺失明显,髓鞘结构蛋白MBP含量明显减少[(0.395±0.019)vs(0.242±0.016);P<0.01];应用图像分析和统计学分析发现,成熟少突胶质细胞的胞体(CC-1+)和(MBP+)呈平行性减少[(42.62±2.48)vs(25.04±3.68)/100μm2;P<0.05];在实验监测过程中,正常小鼠体质量呈上升增加,而CPZ组小鼠体质量先急剧下降,后趋于平稳并稍有恢复;同时行为学实验结果表明,小鼠的运动协调能力、自发活动能力都有明显减弱[(156.17±22.17)vs(78.50±7.80);(240.91±29.52)vs(163.31±21.05);P<0.05],但是对小鼠的探索习性并无明显影响(3.7%/5.4%vs 3.6%/5.3%;P>0.05)。结论利用Cuprizone诱导小鼠脱髓鞘改变与少突胶质细胞减少有关,其主要影响小鼠的运动协调能力和自发活动而不影响探索习性等高级功能。
Objective To investigate the changes of oligodendrocyte in corpus callosum induced by Cuprizone (CPZ) induced demyelination in C57BL / 6 mice and to explore the regulation of the motor ability in mice. Methods 6-week-old male mice fed with normal diet served as control. The experimental group were fed 6-week-old male mice with 0.2% Cuprizone for 6 weeks to induce central nervous system demyelination. LFB histochemistry and MBP immunohistochemistry were used to identify demyelination of the corpus callosum area to confirm the stability of the model. Combined with MBP and CC-1 immunofluorescence double labeling technique, the changes of oligodendrocytes in demyelinated corpus callosum were observed. The whole functional status of the mice was observed by the monitoring of body weight, and the behavioral control effects and characteristics of the mice were observed by behavioral experiments (rotating bar test and open-field test). Results After treatment with Cuprizone for 6 weeks, the myelin deficits in the corpus callosum area were significantly reduced and the MBP content in the myelin sheath was significantly decreased ([0.395 ± 0.019] vs (0.242 ± 0.016), P <0.01] (42.62 ± 2.48) vs (25.04 ± 3.68) / 100μm2, respectively; P <0.05 (P <0.05). The results of image analysis and statistical analysis showed that the cell bodies (CC- ]; In the process of experimental monitoring, normal mice showed an increase in body weight, and CPZ mice first sharp decline in body mass, then tends to be stable and slightly restored; the same time, behavioral experiments show that the mouse’s motor coordination (156.17 ± 22.17) vs (78.50 ± 7.80); (240.91 ± 29.52) vs (163.31 ± 21.05), respectively; P <0.05], but had no significant effect on the exploratory behavior of mice 3.7% / 5.4% vs 3.6% / 5.3%; P> 0.05). Conclusions The use of Cuprizone induced demyelination in mice is associated with a decrease in oligodendrocytes, which mainly affects motor coordination and spontaneous activity in mice without affecting advanced functions such as exploratory behavior.