花椒油对HaCaT细胞膜流动性及膜电位的影响及其机制研究

目的：本文选择HaCaT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HaCaT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HaCaT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HaCaT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HaCaT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。

Effect of Zanthoxylum Bungeanum Oil on HaCaT Cell Membrane Fluidity, Membrane Potential and Its Mechanisms

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 cells, and to investigate the effect of Z. bungeanum oil on intracellular Ca²⁺. The ultramicro Ca²⁺-ATPase kit was used in the determination of Ca²⁺-ATPase activity within HaCaT cells. 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, decrease HaCaT cell membrane potential. Meanwhile, it reduced Ca²⁺-ATPase activity, increased the concentration of intracellular Ca²⁺ within HaCaT cells to influence the Ca²⁺ balance. It was concluded that Z. bungeanum oil may alter the intracellular Ca²⁺ balance to influence the membrane fluidity and potential, in order to increase the epidermal fluidity, reduce the epidermal barrier effect, and increase the drug transdermal absorption.