艾片与麝香酮芳香开窍作用机制研究

Mechanism Research of Aromatics Borneol and Muscone

目的:研究艾片与麝香酮对血脑屏障体外模型MDCK和MDCK-MDR1细胞膜流动性、膜电位、钠钾ATP酶(Na+,K+-ATPase)活性和细胞内游离钙离子浓度的影响,探讨中药芳香开窍的作用机制。方法:体外培养MDCK、MDCK-MDR1细胞。选择对数生长期的细胞,以栀子苷50μg/mL作为对照,与不同浓度艾片与麝香酮配伍干预3 h,采用激光共聚焦显微镜测定细胞膜流动性,流式细胞仪检测细胞膜电位及细胞内游离钙离子浓度的变化,试剂盒检测Na+,K+-ATPase活性。结果:与栀子苷50μg/mL比较,配伍艾片(55.6、111.2μg/mL)和麝香酮(8.34、16.68μg/mL)后可以提高MDCK和MDCK-MDR1细胞膜流动性;配伍艾片(27.8、55.6、111.2μg/mL)和麝香酮(4.17、8.34、16.68μg/mL)可以降低MDCK和MDCK-MDR1细胞膜电位;艾片可使MDCK细胞内钙离子浓度升高,但可降低MDCK-MDR1细胞内钙离子浓度,麝香酮对MDCK和MDCK-MDR1细胞内钙离子浓度具有升高作用;配伍艾片和麝香酮能提高MDCK和MDCK-MDR1细胞Na+,K+-ATPase活性。结论:艾片与麝香酮具有芳香开窍作用,其作用机制可能与其提高细胞膜流动性、Na+,K+-ATPase活性、降低细胞膜电位以及调节细胞内钙离子浓度有关。

Objective: To study the effects of borneol and muscone on membrane fluidity,membrane potential,Na+,K+-ATPase activity and calcium ions of blood brain barrier(BBB) model cells(MDCK and MDCK-MDR1) for exploring the mechanism of aromatics. Methods: MDCK and MDCK-MDR1 cells were incubated and the experiment was performed as following. Cells were incubated with aromatic herbs for 3 h. The membrane fluidity were detected by FRAP. The changes of membrane potentials and the concentration of calcium ions were detected by flow cytometer. Results: Borneol(55. 6,111. 2 μg/mL) and muscone(8. 34,16. 68 μg/mL) significantly enhanced the cell membrane fluidity. Borneol(27. 8,55. 6,111. 2 μg/mL) and muscone(4. 17,8. 34,16. 68 μg/mL) made the MDCK and MDCK-MDR1 membrane potentials less negative or depolarized. Borneol increased the concentration of intracellular free calcium in MDCK while decreased the concentration of intracellular free calcium in MDCK-MDR1 cells. Muscone increased the concentration of calcium in MDCK and MDCK-MDR1 cells. Na+,K+-ATPase activity was significantly increased in borneol and muscone group. Conclusion: The regulating effect of membrane fluidity,membrane potential,Na+,K+-ATPase activity and calcium ions in MDCK and MDCK-MDR1 cells might be one of the mechanisms of borneol and muscone's BBB opening function.