摘要
评价转基因成肌细胞移植治疗帕金森病（ＰＤ）的疗效。方法移植转酪氨酸羟化酶（ＴＨ）基因的成肌细胞（ｎ＝２４）和未经遗传修饰的成肌细胞（ｎ＝１０）于偏侧ＰＤ鼠损毁侧纹状体。用阿朴吗啡（ＡＰＯ）诱发旋转行为，反向聚合酶链反应（ＲＴ－ＰＣＲ）、ＴＨ免疫组化检测ＴＨ基因的表达和ＴＨ蛋白的合成以及高效液相色谱仪－电化学检测器（ＨＰＬＣ－ＥＣＤ）检测纹状体多巴胺及代谢产物含量以此评估基因治疗的效果。结果治疗组移植治疗后ＡＰＯ诱发的旋转行为明显改善（Ｐ＜０．０１），且可持续１３个月，而对照组ＡＰＯ诱发的旋转行为无改善（Ｐ＞０．０５），应用ＲＴ－ＰＣＲ、ＴＨ免疫组化和ＨＰＬＣ－ＥＣＤ在治疗组移植部位检测到ＴＨ基因的表达、ＴＨ蛋白的合成和移植侧纹状体部多巴胺及其代谢产物含量增高。结论遗传修饰的成肌细胞能够在体内长时间、有效地表达ＴＨ，并改善ＰＤ鼠的病理行为，是ＰＤ基因治疗的合适靶细胞之一。
Objective To evaluate the effect of intracerebral grafting of the genetically modified myoblasts expressing tyrosine hydroxylase (TH) on Parkinsonian rats. Methods Myoblasts infected with TH ( n =24) and uninfected myoblast ( n =10) were grafted separatly into the lesioned striatum of rats with unilateral 6 OHDA lesions of nigrostriatal pathway. After grafting, the rotation rate was measured as an index of behavior recovery, and the expression of TH gene was detected by reverse transcription polymerase chain reaction (RT PCR),immunohistochemistry and HPLC ECD. Results Only the animal received the transplantation of genetically modified myoblasts expressing TH displayed a partial reversal of APO induced turning behavior ( P <0.01). Behavioral recovery was maintained for 13 months after grafting. The elevation of TH activity and dopamine content were noted in the lesioned striatum. And the expression of TH mRNA and its immunoactivity were detected.Conclusion The genetically modified myoblasts expressing TH is an attractive candidate for gene therapy of Parkinson disease.
出处
《中华神经科杂志》
CAS
CSCD
1997年第4期201-204,共4页
Chinese Journal of Neurology
基金
国家自然科学基金
卫生部优秀青年人才基金
上海市科委青年科技启明星后计划
关键词
震颤性麻痹
基因治疗
转基因成肌细胞
Parkinson disease Gene therapy Genetically Modified myoblast Tyrosine hydroxylase PT PCR
