乳香、没药挥发油促九分散方中生物碱类成分的HaCaT细胞摄取及其机制研究  

A preliminary study on promoting effect of frankincense and myrrh volatile oil on HaCaT cell uptake of alkaloids in Jiufen Powder and its mechanism

作　　者：高玲 黄诗雨 陈丽华 李瑛瑛 管咏梅 刘丽丽 朱卫丰 GAO Ling;HUANG Shi-yu;CHEN Li-hua;LI Ying-ying;GUAN Yong-mei;LIU Li-li;ZHU Wei-feng(Key Laboratory of Modern Preparation of Chinese Materia Medica,Ministry of Education,Jiangxi University of Chinese Medicine,Nanchang 330004,China)

机构地区：[1]江西中医药大学现代中药制剂教育部重点实验室,江西南昌330004

出　　处：《中草药》2021年第8期2357-2364,共8页Chinese Traditional and Herbal Drugs

基　　金：江西省主要学科学术和技术带头人培养计划(20204BCJL22048);江西省自然科学基金项目(20181BAB205078);江西省重大科技研发专项(S2019ZDYFB0027);江西中医药大学校级大学生创新创业训练计划(202010412182)。

摘　　要：目的研究九分散中乳香、没药挥发油促进角质形成细胞HaCaT摄取马钱子碱、士的宁及盐酸麻黄碱的作用及其机制。方法采用CCK-8法检测乳香、没药挥发油单用/药对与九分散方中马钱子碱、士的宁、盐酸麻黄碱配伍对HaCaT细胞存活率的影响;采用液相色谱串联质谱(LC-MS/MS)定量分析法结合BCA蛋白试剂盒测定在乳香、没药挥发油作用下HaCaT细胞对3种生物碱(马钱子碱、士的宁及盐酸麻黄碱)细胞摄取的影响;以DiBAC4(3)为荧光探针,利用流式细胞仪测定乳香、没药挥发油对HaCaT细胞膜电位的影响。结果不同乳香、没药挥发油均对3种生物碱有一定促透作用,且对脂溶性成分马钱子碱、士的宁的促透效果较水溶性成分盐酸麻黄碱好。乳香、没药挥发油分别作用于DiBAC4(3)标记的HaCaT细胞后,可降低HaCaT细胞膜电位,随着乳香、没药挥发油浓度增加,细胞膜电位荧光强度逐渐增强,具有浓度相关性,表现出类似氮酮的作用方式。结论九分散中乳香、没药挥发油均可促进HaCaT细胞摄取3种生物碱,促进机制可能通过影响皮肤表面负电荷而改变皮肤活性表皮屏障作用,从而有利于药物透过皮肤活性表皮层,其具体机制有待进一步探明。Objective To study the effect and mechanism of the volatile oil of frankincense and myrrh in Jiufen Powder(九分散)in promoting keratinocyte HaCaT uptake of brucine,strychnine and ephedrine hydrochloride.Methods The CCK-8 method was used to detect the effect of frankincense and myrrh volatile oil single use/drug pair on the survival rate of HaCaT cells when combined with brucine,strychnine,and ephedrine hydrochloride in Jiufen Powder prescriptions;LC-MS/MS quantified analytical method combined with BCA protein kit was used to determine the influence of the volatile oil of frankincense and myrrh on the uptake of HaCaT cells to three alkaloid cells;DiBAC4(3)was used as a fluorescent probe to determine the effect of the volatile oil of frankincense and myrrh on HaCaT cell membrane potential.Results The volatile oils of different frankincense and myrrh all had a certain penetration promoting effect on the three alkaloids,and the penetration promoting effect on the lipid-soluble brucine and strychnine was better than the water-soluble ephedrine hydrochloride.After the volatile oil of frankincense and myrrh acted on the HaCaT cells labeled with DiBAC4(3),the volatile oil of frankincense and myrrh could reduce the membrane potential of HaCaT cells.As the concentration increased,the fluorescence intensity of the cell membrane potential increased and showed a significant concentration dependence,showing a similar mode of azone action.Conclusion Both frankincense and myrrh volatile oil can promote the uptake of three alkaloids by HaCaT cells.The promotion mechanism may change the active epidermal barrier function of the skin by affecting the negative charge on the skin surface,thereby facilitating the penetration of the drug through the active epidermal layer.The mechanism needs to be further explored.

关 键 词：九分散 乳香挥发油 没药挥发油 马钱子碱 士的宁 盐酸麻黄碱 细胞膜电位 

分 类 号：R285[医药卫生—中药学]