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目的观察星形胶质细胞在不同年龄(青壮年及中老年)cuprizone(CPZ)脱髓鞘小鼠模型中的激活特点,探讨星形胶质细胞激活与髓鞘脱失可能存在的内在联系。方法选取6周龄(青壮年)及9月龄(中老年)雄性C57BL/6小鼠建立CPZ导致的急性脱髓鞘动物模型,经0.2%CPZ处理6周后,通过体质量测量及卢卡斯快蓝(luxol fast blue,LFB)染色观察模型建立是否成功,利用免疫组织化学染色检测实验组(CPZ)和对照组(CTL)小鼠胼胝体(corpus callosum,CC)及皮层(cortex,CTX)区胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)和髓鞘碱性蛋白(myelin basic protein,MBP)表达变化,并通过Western blot方法进一步检测GFAP及MBP蛋白表达差异。结果 0.2%CPZ处理6周后,青壮年CPZ小鼠于胼胝体及皮层区GFAP阳性细胞数显著增多(P<0.01),同时伴有显著的MBP表达下降(P<0.01);中老年小鼠于胼胝体区GFAP阳性细胞数增多较明显(P<0.05),并伴有MBP表达明显下降(P<0.05);GFAP阳性细胞数的增多和MBP表达下降均表现为青壮年组更加明显(P<0.01)。结论星形胶质细胞的激活于青壮年小鼠更加明显,并与髓鞘损伤呈现出一定的正相关性,星形胶质细胞激活可能是参与髓鞘损伤的重要原因。
Objective To observe the activation of astrocytes in different age (young and middle-aged) cuprizone (CPZ) demyelination mouse models and to explore the possible relationship between astrocyte activation and demyelination. Methods Acute demyelination animal model induced by CPZ was established in male C57BL / 6 mice of 6 weeks old (young and middle-aged) and 9 months old (middle-aged and old). After treated with 0.2% CPZ for 6 weeks, The establishment of the model of luxol fast blue (LFB) staining was observed successfully. The corpus callosum (CC) and cortex (CTX) were detected by immunohistochemical staining in the mice of experimental group (CPZ) and control group (CTL) The changes of glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP) were detected by Western blot. The difference of GFAP and MBP protein expression was further detected by Western blot. Results After 6 weeks of CPZ treatment with 0.2% CPZ, the number of GFAP positive cells in the corpus callosum and cortex of CPZ mice increased significantly (P <0.01), accompanied by significant decrease of MBP expression (P <0.01) The number of GFAP positive cells in the corpus callosum area increased more significantly (P <0.05), accompanied with the decrease of MBP expression (P <0.05). The increase of GFAP positive cells and the decrease of MBP expression were more obvious in young and middle-aged patients (P <0.01) ). Conclusion The activation of astrocytes in young and middle-aged mice is more obvious and shows some positive correlation with myelin damage. Activation of astrocytes may play an important role in the involvement of myelin damage.