花椒油对HaCaT细胞膜流动性及膜电位的影响及其机制研究

Effect of Zanthoxylum Bungeanum Oil on Ha Ca T Cell Membrane Fluidity, Membrane Potential and Its Mechanisms

目的:本文选择Ha Ca T角质形成细胞测定花椒挥发油对其细胞膜流动性和膜电位的影响及其机制,研究花椒挥发油对皮肤活性表皮的影响,探讨其促进药物透皮吸收的作用机制。方法:采用CCK-8试验测定花椒挥发油的细胞毒性,利用荧光漂白恢复技术(FRAP)考察不同浓度花椒挥发油对细胞膜流动性的影响,采用流式细胞仪测定不同浓度花椒挥发油对细胞膜电位的改变,同时考察了花椒挥发油对Ha Ca T细胞内Ca2+浓度的影响,采用超微量Ca2+-ATP酶试剂盒测定花椒挥发油对Ha Ca T细胞内Ca2+-ATP酶活性的影响。结果:相对于常用化学促透剂氮酮,花椒挥发油具有较低细胞毒性,花椒挥发油可增加Ha Ca T细胞膜流动性、降低细胞膜电位,同时可降低Ca2+-ATP酶活性、增加Ha Ca T细胞内Ca2+浓度而影响细胞Ca2+平衡。结论:花椒挥发油可能通过改变细胞内Ca2+平衡而影响细胞膜流动性及膜电位,增加活性表皮流动性以降低皮肤表皮屏障作用,从而利于药物的透皮吸收。

This study was aimed to investigate the effect of essential oil from Zanthoxylum bungeanum Maxim. （Z. bungeanum oil） on cell membrane fluidity and cell membrane potential of HaCaT cells, in order to discuss its influence on epidermis and its action mechanism on medication transdermal penetration enhancer. The cytotoxicity of Z. bungeanum oil was measured by CCK-8 assay. The fluorescence recovery after photobleaching （FRAP） technique was employed to measure changes of membrane fluidity by different concentrations of Z. bungeanum oil. The flow cytometry was used to study the alteration of membrane potential of HaCaT ceils, and to investigate the effect of Z. bungeanum oil on intracellular Ca^2＋. The ultramicro Ca^2＋-ATPase kit was used in the determination of Ca^2＋-ATPase activity within HaCaT eells. The results showed that Z. bungeanum oil possessed low cytotoxicity in comparison to the well-established and standard penetration enhancer Azone. Z. bungeanum oil can enhance HaCaT cell membrane fluidity, deerease HaCaT cell membrane potential. Meanwhile, it redueed Ca^2＋-ATPase activity, increased the concentration of intracellular Ca^2＋ within HaCaT cells to influence the Ca^2＋ balance. It was concluded that Z. bungeanum oil may alter the intracellular Ca^2＋ balance to influence the membrane fluidity and potential, in order to increase the epidermal fluidity, reduee the epidermal barrier effect, and increase the drug transdermal absorption.