Chen Lei, Tian Linyu, Yang Tianhua, Cheng Xinwang, Hermann Stefan, Zhou Dong
Department of Neurology, West China Hospital, Sichuan University, Chengdu, Sichuan, People's Republic of China.
Cell Mol Neurobiol. 2008 Dec;28(8):1057-66. doi: 10.1007/s10571-008-9283-0. Epub 2008 Apr 25.
Over-expression of P-glycoprotein (Pgp), a protein responsible for multidrug resistance (MDR), is responsible for general resistance to anti-epileptic drugs (AEDs). We explored the potential use of gene therapy with adenoviral-delivered RNA interference against mdr1b as a method to sensitize refractory epilepsy to AEDs. We constructed replication-deficient recombinant adenovirus Adeno-mdr1b1 carrying short hairpin RNA (shRNA) targeting against mdr1b, and successfully infected the established Sprague-Dawley rat astrocyte model of Coriaria Lactone-induced Pgp over-expression. The expression levels of mdr1b and Pgp and the Rhodamine123 efflux ratio in trial groups were significantly lower than that of blank control (P < 0.05) during the first 7 days post-infection, with the most inhibition at 48 h. The results suggest that knockdown of MDR using adenovirus not only avoided the toxicity and low rate of plasmid nucleofection, but also overcame its poor efficiency of mdr1b silencing. More importantly, this study may pave the way for a promising approach to remedy refractory epilepsy.
P-糖蛋白（Pgp）是一种导致多药耐药（MDR）的蛋白质，其过度表达会导致对抗癫痫药物（AEDs）产生普遍耐药性。我们探索了利用腺病毒介导的针对mdr1b的RNA干扰进行基因治疗，作为使难治性癫痫对AEDs敏感的一种方法。我们构建了携带靶向mdr1b的短发夹RNA（shRNA）的复制缺陷型重组腺病毒Adeno-mdr1b1，并成功感染了已建立的马桑内酯诱导Pgp过度表达的Sprague-Dawley大鼠星形胶质细胞模型。感染后前7天，试验组中mdr1b和Pgp的表达水平以及罗丹明123外排率均显著低于空白对照组（P<0.05），在48小时时抑制作用最强。结果表明，利用腺病毒敲低MDR不仅避免了质粒核转染的毒性和低转染率，还克服了其对mdr1b沉默效率低下的问题。更重要的是，本研究可能为治疗难治性癫痫开辟一条有前景的途径。
