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。
目的考察薄荷油作为天然经皮促透剂对Ha Ca T皮肤角质形成细胞膜流动性及膜电位的影响,探讨薄荷挥发油经皮促透作用机制。方法采用细胞增殖-毒性试验(CCK-8)测定薄荷挥发油的细胞毒性,利用荧光漂白恢复技术(FRAP)考察不同浓度薄荷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薄荷挥发油对细胞膜电位的改变和对Ha Ca T细胞内Ca2+浓度的影响,采用超微量CA2+-ATP酶试剂盒测定薄荷油对Ha Ca T细胞内Ca2+-ATP酶活性的影响。结果与常用化学促透剂氮酮相比,薄荷挥发油具有较低细胞毒性。薄荷挥发油可增加Ha Ca T细胞膜流动性、降低细胞膜电位,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。结论薄荷挥发油可能通过影响皮肤表皮细胞Ca2+平衡而改变细胞膜流动性及膜电位,降低皮肤屏障作用,从而促进药物的透皮吸收。
Our previous studies had confirmed that the essential oil from Zanthoxylum bungeanum Maxim. （Z. bungeanum oil） could effectively enhance the percutaneous permeation of drug molecules as a natural transdermal penetration enhancer. The aim of the present study is to investigate and compare the skin penetration enhancement effect of Z. bungeanum oil and its main components on traditional Chinese medicine （TCM） active components. Toxicities of Z. bungeanum oil and three selected terpene compounds （terpinen-4-ol, 1,8-cineole, and limonene） in epidermal keratinocytes （HaCaT） and dermal flbroblast （CCC-ESF-1） cell lines were measured using an MTT （3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide） assay. Five model drugs in TCM external preparations, namely osthole （OT）, tetramethylpyrazine （TMP）, ferulic acid （FA）, puerarin （PR）, and geniposide （GP）, which were selected based on their lipophilicity denoted by IogKo^w, were tested using in vitro permeation studies in which vertical Franz diffusion ceils and rat abdominal skin were employed. The secondary structure changes of skin stratum corneum （SC） and drug thermodynamic activities were investigated to understand their mechanisms of action using Fourier transform infrared （FTIR） spectroscopy and saturation solubility studies, respectively. It was found that Z. bungeanum oil showed lower toxicities in both HaCaT cells and CCC-ESF-1 cells compared with three terpene compounds used alone. The enhancement permeation capacities by all tested agents were in the following increasing order： terpinen-4-ol=1,8-cineole〈limonene〈Z, bungeanum oil. The mechanisms of permeation enhancement suggested that these enhancers promoted the skin permeation of drugs mainly by affecting SC lipids. These results indicated that Z. bungeanum oil exhibited better performance in enhancing the skin permeation of active components in TCM preparations.
Yi LANHui LIYan-yan CHENYe-wen ZHANGNa LIUQing ZHANGQing WU
