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目的:观察星形胶质细胞在帕金森病(Parkinson’s disease,PD)模型大鼠脑室下区、中脑的分布并进行PD模型大鼠脑室下区星形胶质细胞的体外培养、纯化和鉴定。方法:采用免疫荧光方法观察胶原纤维酸性蛋白(glial fibrillary acidic protein,GFAP)在PD模型大鼠中脑、脑室下区的表达;取PD模型大鼠侧脑室下区细胞进行体外培养、传代、贴壁分化和免疫荧光法鉴定。结果:(1)GFAP阳性细胞在PD模型大鼠中脑病侧较健侧明显增生,且GFAP荧光定量强度值在病侧较健侧明显增大(P<0.05);GFAP阳性细胞在侧脑室、第三脑室和第四脑室室周区、视上核、下丘脑室旁核外侧大细胞部、正中隆起大量分布;(2)PD模型大鼠侧脑室下区培养分化的细胞具有星形胶质细胞的典型形态,其中原浆性星形胶质细胞突起粗短,分支多;纤维性星形胶质细胞突起细长,分支较少。结论:(1)星形胶质细胞在PD模型大鼠中脑和脑室下区的分布有明显区域性,可能与病侧的病理性增生相关,与相应区域的生理活动、调节功能及有关;(2)取PD模型大鼠侧脑室下区进行体外培养、分化获得星形胶质细胞的实验方法可靠。
OBJECTIVE: To observe the distribution of astrocytes in the subventricular zone and midbrain of Parkinson’s disease (PD) model rats and to culture, identify and purify the astrocytes in the subventricular zone of rat model of PD . Methods: Immunofluorescence was used to observe the expression of glial fibrillary acidic protein (GFAP) in the midbrain and subventricular zone of PD model rats. The cells in the subventricular zone of PD model rats were cultured, passaged and pasted Wall differentiation and immunofluorescence identification. Results: (1) GFAP-positive cells proliferated more significantly in the midbrain than in the contralateral side in PD model rats, and the GFAP fluorescence intensity was significantly increased in the diseased side as compared with the contralateral side (P <0.05); GFAP positive cells in the lateral ventricle, The third ventricle and the fourth ventricle of the periventricular area, as the supraoptic nucleus, the hypothalamic paraventricular nucleus outside the large cell, the median bulge distributed; (2) PD model rat subventricular zone cultured differentiated cells with astrocytes The typical cell morphology, in which prothoracic astrocytes protruding stubby, branching and more; fibrous astrocytes protruding slender, fewer branches. Conclusion: (1) The distribution of astrocytes in the midbrain and the subventricular zone of PD model rats is obviously regional, which may be related to the pathological hyperplasia of the diseased side and related to the physiological activities and regulatory functions in the corresponding area. (2) The PD model rats were cultured in vitro in the subventricular zone, and the experimental method of obtaining astrocytes by differentiation was reliable.