萜烯类经皮促透剂对皮肤活性表皮层的影响及其机制研究(1)
[摘要]该文选择HaCaT皮肤角质形成细胞测定萜烯类经皮促透剂对皮肤活性表皮层的影响，探讨其促进药物透皮吸收的作用机制。选择具良好经皮促透效果的萜烯类化合物，即薄荷醇、柠檬烯、1，8-桉树脑、薄荷酮、4-萜品醇和长叶薄荷酮;采用MTT试验测定萜烯类经皮促透剂的细胞毒性，利用荧光漂白恢复技术(FRAP)考察不同萜烯类促透剂对细胞膜流动性的影响，采用流式细胞仪测定不同萜烯类促透剂对细胞膜电位的改变，同时考察了萜烯类促透剂对HaCaT细胞内Ca2+浓度的影响，采用超微量Ca2+-ATP酶试剂盒测定萜烯促透剂对HaCaT细胞内Ca2+-ATP酶活性的影响。实验结果表明，相对于常用化学促透剂氮酮，6种萜烯类促透剂均显示较低细胞毒性;所选择的萜烯类促透剂可显著增加HaCaT细胞膜流动性、降低细胞膜电位，同时可降低HaCaT细胞Ca2+-ATP酶活性和细胞内Ca2+浓度而影响细胞Ca2+平衡。因此，萜烯类促透剂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，增加皮肤活性表皮流动性而降低皮肤屏障作用，从而利于药物的透皮吸收。
[关键词]萜烯类促透剂;HaCaT细胞;细胞膜电位;细胞膜流动性;Ca2+;经皮促透剂
, 百拇医药
Effect of terpene penetration enhancer and its mechanisms on membrane
fluidity and potential of HaCaT keratinocytes
LAN Yi， WANG Jing-yan， LIU Yan， RU Qing-guo， WANG Yi-fei， YU Jing-xin， WU Qing*
(School of Chinese Materia Medica， Beijing University of Chinese Medicine， Beijing 100102， China)
[Abstract]The aim of this paper was to investigate the effect of terpene penetration enhancers on membrane fluidity and membrane potential using HaCaT keratinocytes， and study the potential mechanisms of these terpene compounds using as natural transdermal penetration enhancer. Six terpene compounds， namely menthol， limonene， 1，8-cineole， menthone，terpinen-4-ol and pulegone， were chosen in this study on account of their good penetration-enhancement activities. The cytotoxicity of these terpene compounds was measured using an MTT assay. The fluorescence recovery after photobleaching (FRAP) technique was employed to measure the change of membrane fluidity of HaCaT cells. The flow cytometer was used to study the alteration of membrane fluidity of HaCaT cells， and investigate the effect of terpene compounds on intracellular Ca2+. It was found that 6 terpene compounds possessed low cytotoxicity in comparison to the well-established and standard penetration enhancer azone. Those terpene compounds could significantly enhance HaCaT cells membrane fluidity and decrease HaCaT cells membrane potentials. Meanwhile， after treated with various terpene compounds， the Ca2+-ATPase activity and intracellular Ca2+ of HaCaT cells was decreased significantly. Terpene penetration enhancers perhaps changed the membrane fluidity and potentials of HaCaT cells by altering the Ca2+ balance of the cell inside and outside， resulting in the low skin permeability to increase the drug transdermal absorption.
[Key words]terpene penetration enhancer; HaCaT; membrane fluidity; membrane potential; Ca2+; transdermal penetration enhancer
doi：10.4268/cjcmm20150414, 百拇医药(兰颐 王景雁 刘艳 茹庆国 王逸飞 于竞新 吴清)
[关键词]萜烯类促透剂;HaCaT细胞;细胞膜电位;细胞膜流动性;Ca2+;经皮促透剂
, 百拇医药
Effect of terpene penetration enhancer and its mechanisms on membrane
fluidity and potential of HaCaT keratinocytes
LAN Yi， WANG Jing-yan， LIU Yan， RU Qing-guo， WANG Yi-fei， YU Jing-xin， WU Qing*
(School of Chinese Materia Medica， Beijing University of Chinese Medicine， Beijing 100102， China)
[Abstract]The aim of this paper was to investigate the effect of terpene penetration enhancers on membrane fluidity and membrane potential using HaCaT keratinocytes， and study the potential mechanisms of these terpene compounds using as natural transdermal penetration enhancer. Six terpene compounds， namely menthol， limonene， 1，8-cineole， menthone，terpinen-4-ol and pulegone， were chosen in this study on account of their good penetration-enhancement activities. The cytotoxicity of these terpene compounds was measured using an MTT assay. The fluorescence recovery after photobleaching (FRAP) technique was employed to measure the change of membrane fluidity of HaCaT cells. The flow cytometer was used to study the alteration of membrane fluidity of HaCaT cells， and investigate the effect of terpene compounds on intracellular Ca2+. It was found that 6 terpene compounds possessed low cytotoxicity in comparison to the well-established and standard penetration enhancer azone. Those terpene compounds could significantly enhance HaCaT cells membrane fluidity and decrease HaCaT cells membrane potentials. Meanwhile， after treated with various terpene compounds， the Ca2+-ATPase activity and intracellular Ca2+ of HaCaT cells was decreased significantly. Terpene penetration enhancers perhaps changed the membrane fluidity and potentials of HaCaT cells by altering the Ca2+ balance of the cell inside and outside， resulting in the low skin permeability to increase the drug transdermal absorption.
[Key words]terpene penetration enhancer; HaCaT; membrane fluidity; membrane potential; Ca2+; transdermal penetration enhancer
doi：10.4268/cjcmm20150414, 百拇医药(兰颐 王景雁 刘艳 茹庆国 王逸飞 于竞新 吴清)