Dopaminergic trophism after intrastriatal injection of lentivirus-transferred GDNF in Parkinson rat model.

Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica Pub Date : 2003-10-01

Bing Sun, Guo-Zhen Hui, Li-He Guo, Jakob Reiser

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Abstract

To investigate the effects of lentivirus-mediated transfection of GDNF on Parkinson s disease (PD). The pNL-gdnf plasmid was constructed by replacing the LacZ-coding region present in pNL-lacZ/CMV. Vector particles involved a three-plasmid lentivirus expression system were co-transferred into 293T cells through calcium phosphate method. High-titer virus was collected from infected 293T cells and injected into lesion-side striatum of PD rats, and their apomorphine-induced rotations were assayed at day 14, 30 and 60, respectively. GDNF protein was detected by Western blot analysis, and the expression of lacZ and TH were detected by immunochemistry. Results showed that behavioral recovery gradually appeared after transplantation, and a significant reduction in the rotational response was observed at the 14th day. Meanwhile, gdnf expression maintained for at least 60 d, which had dopaminergic trophism to a certain degree, indicating that lentivirus-mediated transfection gdnf could effectively improve the clinical function of PD rats, which provide a potential attractive tool of gene therapy for PD.

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慢病毒转移GDNF在帕金森模型大鼠纹状体内注射后多巴胺能的营养作用。

目的探讨慢病毒介导的GDNF转染对帕金森病(PD)的影响。pNL-gdnf质粒通过替换存在于pNL-lacZ/CMV中的lacz编码区构建。采用磷酸钙法将慢病毒三质粒表达系统的载体颗粒共转移到293T细胞中。从感染的293T细胞中收集高滴度病毒，注射到PD大鼠病变侧纹状体中，分别在第14、30和60天检测阿帕吗啡诱导的旋转。Western blot法检测GDNF蛋白表达，免疫化学法检测lacZ、TH蛋白表达。结果显示，移植后行为逐渐恢复，第14天旋转反应明显降低。同时，gdnf表达维持至少60 d，具有一定程度的多巴胺能营养作用，说明慢病毒介导转染gdnf可有效改善PD大鼠的临床功能，为PD基因治疗提供了一种潜在的有吸引力的工具。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica

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