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探讨慢病毒介导GDNF对帕金森病的治疗作用。将gdnf片段替代pNL lacZ CMV质粒中的LacZ编码区 ,构建pNL gdnf质粒。采用磷酸钙转染方法 ,将慢病毒系统中三个质粒瞬时共转染 2 93T细胞 ,并收集病毒粒子。用立体定位仪将高滴度病毒注射入PD大鼠的纹状体中。分别在治疗后 14 ,30 ,6 0天检测阿朴吗啡 (APO)诱导旋转反应的变化 ;用Western印迹方法测定蛋白质的表达 ;用免疫组织化学方法检测lacZ和TH的表达 ;移植治疗后PD鼠的行为学逐渐有了改善 ,尤以治疗后14天明显。GDNF蛋白在大鼠脑内至少表达了 6 0天 ,对多巴胺神经有一定的神经营养作用。因此 ,慢病毒介导GDNF能显著改善PD鼠的行为学表现 ,是治疗帕金森病的有效方法之一。
To investigate the lentivirus-mediated GDNF treatment of Parkinson’s disease. The gdnf fragment was substituted for the LacZ coding region in the pNL lacZ CMV plasmid to construct the pNL gdnf plasmid. Three plasmids were transiently cotransfected into 293T cells using the calcium phosphate transfection method and virus particles were collected. High-titer virus was injected into the striatum of PD rats using a stereotaxic instrument. The apomorphine (APO) -induced rotational response was detected at 14, 30 and 60 days after treatment respectively. The protein expression was detected by Western blotting. The expression of lacZ and TH was detected by immunohistochemistry. Behavioral learning gradually improved, especially after 14 days of treatment. GDNF protein was expressed in rat brain for at least 60 days and had certain neurotrophic effect on dopamine nerve. Therefore, lentivirus-mediated GDNF can significantly improve the behavioral performance of PD mice is one of the effective ways to treat Parkinson’s disease.