冰片对HaCaT细胞膜流动性及膜电位的影响

摘要

Objective To investigate the action mode of borneol on activity of epidermal skin;To investigate action mode of borneol as penetration enhancer. Methods The well-established and standard penetration enhancer Azone was employed as a positive control in this study. The cytotoxicities of borneol and Azone on HaCaT cells were detected by CCK-8 assay, and their half 50% inhibitory concentrations (IC50) were calculated. The fluorescence recovery after photo bleaching was employed to investigate the effect of borneol and Azone on membrane fluidity, and the flow cytometer was used to monitor the changes of membrane potential of HaCaT cell after treated with these penetration enhancers. Results The IC50 values of borneol and Azone were 2.826 , 0.172 mmol/L, respectively. Borneol could significantly improve the membrane fluidity in a concentration-dependent manner, and effectively decrease the membrane potential of HaCaT cell, which exhibited the performances similar to those of Azone. Conclusion The penetration enhancement mechanism of borneol was associated with the concentrations of Ca2+ in keratinocytes, which changes the membrane fluidity and membrane potential of HaCaT cell.%目的：研究冰片对皮肤活性表皮作用方式，探究冰片作为经皮促透剂的促透作用机制。方法选择常用经典化学经皮促透剂氮酮作为阳性对照促透剂，采用CCK-8试剂盒测定并计算冰片对HaCaT细胞的半数抑制率(IC50)，利用荧光漂白恢复技术测定不同浓度冰片对HaCaT细胞膜流动性的影响，采用流式细胞仪测定冰片对HaCaT细胞膜电位及细胞膜内Ca2+浓度的影响。结果冰片和氮酮的药物IC50分别为2.826、0.172 mmol/L；冰片可增加HaCaT细胞膜流动性，且随着药物浓度的增大而增加，呈剂量依赖性关系；冰片可降低HaCaT细胞膜电位和HaCaT细胞内Ca2+浓度，表现出类似氮酮作用特点。结论冰片的经皮促透作用机制可能与其影响角质细胞内Ca2+浓度进而改变活性表皮角质形成细胞膜电位和膜流动性有关。