RNA interference inhibiting permeability glycoprotein over-expression in pharmaco-resistance rat astrocytes model
- VernacularTitle:核糖核酸干扰抑制P-糖蛋白在大鼠耐药星形胶质细胞中过度表达
- Author:
Lei CHEN
;
Linyu TIAN
;
Tianhua YANG
;
Dong ZHOU
- Publication Type:Journal Article
- Keywords:
Adenoviridae;
RNA,small interference;
P-glycoprotein;
Astrecytes;
Transfection
- From:
Chinese Journal of Neurology
2008;41(10):699-703
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the effect of adenoviral-delivered short hairpin RNA (shRNA) target against permeability glycoprotein (Pgp) as a new drug in anti-epileptic drug resistance epilepsy treatment and to evaluate its efficiency. Methods MDR Sprague-Dawley (SD) rat estrocyte model was induced by Coriaria Lactone (CL), mainly over-expressing mdrlb. To reverse the drug resistance, astrecytes were treated with constructed replication deficient adencvirus AdS-EGFP-shRNAI-U6 delivering short hairpin (shRNA) target agianst mdrlb gene. Total RNA and protein were extracted from the infected cells, mdr1 b level was detected by Quantitative Real-time PCR whereas Pgp by Western blot, Rhodamine123 (Rho123) efflux ratio by Flow Cytometry. Results AdS-EGFP-shRNA1-U6 was succesfully constucted with high virus titer of 6×1010 pfu/ml. The interference efficency of AdS-EGFP-shRNA1-U6 agianst mdrlb in rat astrecyte model was about 94%. The Rho123 efllux ratio was about 15. 8%, significiently lower than control group which was 56. 2% (F = 127.5, P < 0. 05). Conclusions Pgp over-expression has been successfully suppressed and MDR has been reversed, which may provide a premising approach for refractory epilepsy remedy.
