马桑内酯体外诱导大鼠星形胶质细胞P-糖蛋白表达

Coriaria Lactone Induces in vitro the Overexpression of Multidrug Resistance P-glycoprotein in Astrocyte from Rat Brain

目的马桑内酯体外诱导耐抗癫痫药物(AED)的星形胶质细胞,并探讨可能的耐药机制。方法培养新生大鼠的星形胶质细胞,传代后向其培养液中按体积1μL,3μL,9μL,15μL滴加马桑内酯,分别在加入药物后第7d,14d,21d,28d用免疫细胞化学和图像分析的方法检测并分析P-糖蛋白的表达。结果从第14d开始至28d,随诱导时间的延长,P-糖蛋白的表达明显增加(P<0.01);在21d,28d组,随马桑内酯剂量的增加,P-糖蛋白的表达无明显增加(P>0.05)。结论马桑内酯体外诱导的大鼠星形胶质细胞对AEDs有耐药性,P-糖蛋白的表达增强可能是其耐药性产生的重要机制。

Objective The aim of this study was to explore the possible pharmacoresistant mechanism by developing a kind of astrocyte model induced by Coriaria lactone （CL）. Methods Astrocyte cells from natal Sprague-Dawley rats （within 24 hours of birth） were cultured. Different volumes of CL（1 μL,3 μL, 9 μL, 15 μL） were added to the cultures for 7, 14, 21 or 28 days. The expression of P-glycoprotein （Pgp）, the protein product of multidrug resistance 1 （MDR1）, was investigated with immunocytochemistry and image analysis. Results CL induced the overexpression of Pgp in astrocytes in a time-dependent manner. The up-regulation of Pgp expression was detected to any concentration of CL from day 14 to 28 （P〈0. 01）. On the day21,28, the up-regulation of Pgp expression level was no dose-dependent manner（P〉0. 05）. Conclusion The study provides a ideal astrocyte model of pharmacoresistant epilepsy, which is induced by CL. The increased expressions of P-gp may involve in induction of the pharmacoresistance.