中国药理学通报2011，Vol.27Issue(2)：234-239,6.
慢病毒介导的新型Tet-On系统大鼠GDNF和TH双基因脑内转移对帕金森病大鼠模型的保护作用
Protective effects of the intracerebral transfer of the lentiviral-mediated GDNF and TH bi-gene on the basis of improved Tet-on system in a rat model of Parkinson's disease
摘要
Abstract
Aim To study the protective effect of lentiviral-mediated rat glial cell line-derived neurotrophic factor( GDNF ) and tyrosine hydroxylase( TH ) on the basis of improved Tet-on system gene transfer on dopaminergic neurons in a rat model of Parkinson's disease( PD ).Methods Recombinant lentivirus( Lv-THGDNF )carrying rat GDNF and TH genes and tetracy cline response element containing mouse albumin gene promoter( Palb ) together with tetracycline-controlled transactivator rtTA2s-M2 virus was injected into the left striate bodies SD rats.After that.the expression of GDNF and TH genes was regulated by doxycycline.One week later, ipsilateral intrastriatal injection of 6hydroxydopamine( 6-OHDA )was to injury dopaminergic neurons.The neuroprotective effects of Lv-TH-GDNF were evaluated by apomorphine-induced rotational behavior.immunohistochemistry assay of the tyrosine hydroxylase positive neurons in the substantia nigra,and the measurement of dopamine level in the striatum by high performance liquid chromatography-electric chemical discharge( HPLC-ECD ).RT-PCR and western blotting were performed to check the expression of Lv-TH-GDNF in the brain.Tests were compared with the PBS control group and the non-injected control group.Results Tests showed apomorphine-induced contralateral turning effect was significantly reduced in Lv-TH-GDNF+ rtTA2s-M2 + DOX-pre-treated rats as compared with PBS-pre-treated rats when measured at 4 weeks post lesion by 6-OHDA( P＜0.01 ), the expression of TH-positive cells in the lesioned substantia nigra and DA levels in the lesioned striatum were significantly higher in Lv-TH-GDNF + rtTA2s-M2 + DOXpre-treated rats than that in PBS-pre-treated rats( P ＜0.01 ) , but both were significantly lower than that of in non-injected rats( P ＜0.01 ).RT-PCR and western blotting showed the expression of exogenous GDNF and TH genes was significantly raised in the lesioned striatum in Lv-TH-CDNF+rtTA2s-M2 + DOX -pre-treated rats as compared with PBS-pre-treated rats.Conclusion The results suggest that lentiviral-mediated rat CDNF and TH bi-gene on the basis of improved Tet-on system ,which is intrastriataly transferred to be induced by doxycycline, can significantly retard progressive degeneration of dopaminergic neurons of nigrostriatal system from 6-OHDA-induced injury, which shows that it has neuroprotective effect on the dopaminergic neurons of PD.关键词
胶质细胞源性神经营养因子/酪氨酸羟化酶/慢病毒/Tet-On系统/活体直接转移/帕金森病分类
医药卫生引用本文复制引用
张阳,张志坚,俞晓岚,黄志新,吴秀丽..慢病毒介导的新型Tet-On系统大鼠GDNF和TH双基因脑内转移对帕金森病大鼠模型的保护作用[J].中国药理学通报,2011,27(2):234-239,6.基金项目
福建省卫生厅医学创新课题(No 2007-CX-15) （No 2007-CX-15）
