Experimental study on expression of TH and GDNF bi-gene of lentivirus by improved Tet-On system for a rat model of Parkinson’ s disease
10.3969/j.issn.1001-1978.2015.02.021
- VernacularTitle:慢病毒介导新型Tet-On系统调控大鼠GDNF和TH双基因表达对帕金森病大鼠的实验研究
- Author:
Yang ZHANG
;
Zhijian ZHANG
;
Xiaolan YU
;
Dongping CHEN
;
Xiuli WU
;
Zhihong WANG
;
Zusheng CHEN
;
Qiuhong CHEN
- Publication Type:Journal Article
- Keywords:
Tet-On system;
TH gene;
GDNF gene;
Parkinson’ s disease;
lentivirus;
rat
- From:
Chinese Pharmacological Bulletin
2015;(2):251-255,256
- CountryChina
- Language:Chinese
-
Abstract:
Aim To investigate TH and GDNF genes expression and regulation of lentivirus ( Lv-TH-GDNF ) based on improved Tet-On system and the effect of the Lv-TH-GDNF intrastriatal transfer on a rat Parkinson’ s disease( PD) model. Methods 1. HeLa cells were infected by obtained Lv-TH-GDNF and rtTA2 s-M2 vi-rus. The expression of tyrosine hydroxylase ( TH ) and glial cell line-derived neurotrophic factor( GDNF) genes was induced by doxycycline( Dox) which was examined by Western blot. 2. The Lv-TH-GDNF together with rtTA2 s-M2 viruses were injected into lesion-side stria-tum of a rat PD model, and the expression of GDNF and TH genes was induced by Dox. Then, the effects of Lv-TH-GDNF were evaluated by the apomorphine-induced rotational behavior, the number of dopaminer-gic neurons in substantia nigra,DA and DOPAC levels in the lesion-side striatum. In addition, Western blot was performed to check the expression of TH and GD-NF genes in the transplanted striatum. Results 1. In vitro studies on HeLa cells, Western blot showed clear protein bands of TH and GDNF in the Dox-positive group, but not in the Dox-negative group. 2. In vivo experiments in animals, the results showed that, 4 weeks after transplantation, the apomorphine-induced turning effect was significantly improved ( P<0 . 01 ) , the number of TH-positive cells in the lesion-side sub-stantia nigra pars compacta as well as the content of DA and DOPAC, the protein level of GDNF and TH genes in the lesion-side striatum was significantly in-creased ( P<0 . 01 ) , each of which was only in Lv-TH-GDNF+rtTA2 s-M2+Dox-treated rats as compared with PBS-treated rats. Conclusion The expression of TH and GDNF genes in Lv-TH-GDNF based on im-proved Tet-On system is effectively regulated by tetra-cycline antibiotics without basal activity in vitro, and the intrastriatal transfer of which has certain therapeutic effect on PD rats.
