目的 体外细胞模型法研究川芎中藁本内酯、洋川芎内酯A和洋川芎内酯Ⅰ对天麻素(GAS)跨膜转运的影响及作用机制。方法 四甲基偶氮唑蓝(MTT)法考察GAS、藁本内酯、洋川芎内酯A和洋川芎内酯Ⅰ的细胞毒性;采用MDCK-MDR1细胞模型分析GAS的跨膜转运，以及藁本内酯、洋川芎内酯A和洋川芎内酯Ⅰ对GAS跨膜转运的影响;Western Blot法研究藁本内酯、洋川芎内酯A和洋川芎内酯Ⅰ对P-gp蛋白表达的影响。结果 GAS在MDCK-MDR1上Papp(A→B)＜1×10-6cm·s-1，属于难跨膜转运的药物;P-gp抑制剂使GAS的Papp(B→A)/Papp(A→B)从1.29减少到0.79，表明GAS的跨膜转运受到了P-gp的外排作用。30 μg·mL-1藁本内酯和120 μg·mL-1洋川芎内酯Ⅰ能极显著性增加GAS的Papp(A→B)(P<0.01)，120 μg·mL-1洋川芎内酯A能显著性增加GAS的Papp(A→B)(P<0.05)。高、中、低剂量的藁本内酯、洋川芎内酯A和洋川芎内酯Ⅰ均能显著性抑制P-gp蛋白表达。结论 GAS跨膜转运机制为被动转运，兼有外排蛋白参与。川芎中的藁本内酯、洋川芎内酯A和洋川芎内酯Ⅰ可通过抑制P-gp蛋白的表达来促进GAS的跨膜转运。
Abstract
OBJECTIVE To study the influence and mechanism of bioactive ingredients of Ligusticum chuanxiong Hort. on the transport of gastrodin based on cell culture model in vitro. METHODS Cell toxicity of gastrodin, ligustilide, senkyunolide Ⅰ and senkyunolide A were detected by MTT assay. The transport mechanism of gastrodin and the influence of ligustilide, senkyunolide Ⅰ and senkyunolide A on the transport of gastrodin were studied in MDCK-MDR1 monolayer cells. The changed expressions of P-gp caused by ligustilide, senkyunolide Ⅰ and senkyunolide A were analyzed by Western blotting.RESULTS Gastrodin showed relatively poor absorption in MDCK-MDR1 cells for its apparent permeability coefficients were less than 1×10-6 cm·s-1. P-gp inhibitor made the Papp(B→A)/Papp(A→B) of gastrodin reduced from 1.29 to 0.79, which indicated that the transport of gastrodin was influenced by P-gp. In the presence of ligustilide(30 μg·mL-1) or senkyunolide Ⅰ(120 μg·mL-1), the Papp(A→B) of gastrodin in MDCK-MDR1 were significantly increased(P<0.01). In the presence of senkyunolide A(120 μg·mL-1), the Papp(A→B) of gastrodin in MDCK-MDR1 were markedly increased(P<0.05). High, medium and low dose of ligustilide, senkyunolide A and senkyunolide Ⅰ could significantly inhibit the expression of P-gp protein.CONCLUSION The results indicate that the transport mechanism of gastrodin might be passive diffusion as the dominating process with the active transportation mediated mechanism involved. Ligustilide, senkyunolide A and senkyunolide Ⅰ increase the transport of gastrodin attribute to down-regulate P-gp expression.
关键词
天麻素 /
P-糖蛋白 /
藁本内酯 /
洋川芎内酯A /
洋川芎内酯Ⅰ /
血脑屏障
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Key words
gastrodin /
p-glycoprotein /
ligustilide /
senkyunolide A /
senkyunolide Ⅰ /
blood-brain barrier
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中图分类号：
R965
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脚注
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基金
国家自然科学基金资助项目(81360645);江西省青年科学家资助项目(20122BCB23021);江西省自然科学基金资助项目(20142BAB205088);江西省教育厅资助项目(GJJ13606)
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