蛋白酶抑制剂Lactacystin建立帕金森病大鼠模型的胶质细胞反应

目的：探讨蛋白酶抑制剂造成的蛋白酶功能损伤的帕金森病（PD）动物模型黑质区胶质细胞的病理特点.方法：将SD大鼠随机分成3组：Lactacystin组、生理盐水组及正常对照组.使用立体定向注射方法向黑质区注射Lactacystin制作PD大鼠单侧模型.采用免疫组化方法观察大鼠黑质区酪氨酸羟化酶（TH）、胶质纤维酸性蛋白（GFAP）、植物凝集素（BSI-B4）免疫阳性细胞并对其进行细胞计数及灰度分析,对黑质区的多巴胺神经元、星形胶质细胞及小胶质细胞进行分析.结果：模型组病变侧黑质区TH阳性细胞较正常组和生理盐水组明显减少（P〈0.05）,胞浆染色浅.GFAP免疫组化染色模型组灰度值较正常组及生理盐水组降低（P〈0.05）.BSI-B4免疫组化染色模型组阳性细胞均数较正常组和生理盐水明显增加（P〈0.05）;模型组灰度值较正常组及生理盐水组低（P〈0.05）.结论：泛素蛋白酶抑制剂lactacystin可导致多巴胺能神经元变性死亡,并可导致黑质区小胶质细胞增生活化与星形胶质细胞的活化.

The reaction of glial cells in the Parkinson's rat model induced by proteinase inhibitor Lactacystin

Objective：To study the pathologic characteristics of the glial cells in the substantia nigra in rat model with Parkinson Disease （PD） induced by proteasome inhibitor. Methods： SD rats were randomly divided into 3 groups： the Lactacystin group, normal saline group and control group. Stereotaxic injection of Lactacystin into substantia nigra was used to establish unilateral PD rat model. Immunohistochemical stain were use to study the change of tyrosine hydroxylase （TH）, glial fibrillary acidic protein （GFAP）, and BSI-B4 induced positive cells in the substantia nigra region in rats, positive cell count and gradation analysis were performed; besides, dopamine neuron, astrocytes and microglia over substantia nigra region were also analyzed. Results： Compared with the control and normal saline group, the TH immunopositive cells decreased significantly （ P 〈 0.05 ） and endoehylema staining became shallower in the lesion lateral of the Lactacystin group ; the Gray-Scale in GFAP immunohistochemistry group was lower,and positive cell count in BSI-IM immunohistochemistry group increased significantly than that of the other two groups （ P 〈 0.05 ） ; the Gray-Scale analysis in model group was lower than that of physical saline group （P 〈 0. 05 ）. Conclusion： Ubiquitin proteinase inhibitor Laetacystin may induce the degeneration of the dopaminergic neuron and might lead to activation of glial cells and astrocytes in the substantia nigra region.