RNA interference inhibiting permeability glycoprotein over-expression in pharmaco-resistance rat astrocytes model / 中华神经科杂志

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.