甘草酸18位差向异构体不同配比和浓度对Caco-2细胞P-gp功能的影响

doi:10.3969/j.issn.1000-1565.2016.03.006

摘要/Abstract

摘要： 研究甘草酸18位差向异构体18α-甘草酸(18α-Gly)与18β-甘草酸(18β-Gly)不同配比和浓度对结肠癌(Caco-2)细胞P-糖蛋白(P-gp)功能的影响,选择最佳浓度比例.建立细胞模型,选择甘草酸总浓度为1∶10∶30∶60∶120∶240 μmol/L,18α-Gly与18β-Gly物质的量比分别为10∶0、8∶2、6∶4、5∶5、4∶6、2∶8、0∶10,根据细胞存活率,选择合适浓度比例,利用流式细胞仪测定细胞内荧光强度,寻找荧光强度最大的浓度比,即对P-gp抑制作用最强,P-gp功能最弱.研究结果表明甘草酸总浓度为1 μmol/L时,随着18α-Gly比例的减少,荧光强度逐渐减弱,P-gp诱导作用逐渐增加.甘草酸总浓度为10 μmol/L和60 μmol/L时,随着两者物质的量比的变化,荧光强度变化明显;当总浓度为10 μmol/L,n(18α-Gly)∶n(18β-Gly)=4∶6时,荧光强度较强,抑制作用明显;当总浓度为60 μmol/L,两者物质的量比为5∶5时荧光强度最强,抑制作用最强.

关键词: 18α-甘草酸, 18β-甘草酸, P-糖蛋白功能, Caco-2细胞, 不同配比/浓度

Abstract: To evaluate the effect of glycyrrhizic acid 18 epimers,18α-glycyrrhizic acid(18α-Gly)and 18β-glycyrrhizic acid(18β-Gly)at different proportion and concentration on the P-glycoprotein(P-gp)activity in colon cancer Caco-2 cell to select their appropriate concentration and proportion in use.Making a cell model,selecting the total glycyrrhizic acid concentration of 1,10,30,60,120,240 μmol/L and proportion of 10∶0,8∶2,6∶4,5∶5,4∶6,2∶8,0∶10,determining their optimum concentration and proportion by cell survival rate,and detecting the maximum fluorescence value by flow cytometer,that is,thestrongest inhibitory effect of P-gp and its weakest function.When the concentration of glycyrrhizic acid was 1 μmol/L,the fluorescence intensity became gradually weak and its induction fuction was strengthed gradually with the reduction in proportion of 18α-Gly.When the concentration of glycyrrhizic acid was 10 μmol/L and- DOI:10.3969/j.issn.1000-1565.2016.03.006甘草酸18位差向异构体不同配比和浓度对Caco-2细胞P-gp功能的影响赵燕燕^1,2,孙东¹,刘丽艳³,柳亚飞¹,王静¹( 1.河北大学化学与环境科学学院,河北保定 071002;2.河北大学药学院,河北保定 071002;3.河北大学医学实验中心,河北保定 071000)摘要:研究甘草酸18位差向异构体18α-甘草酸(18α-Gly)与18β-甘草酸(18β-Gly)不同配比和浓度对结肠癌(Caco-2)细胞P-糖蛋白(P-gp)功能的影响,选择最佳浓度比例.建立细胞模型,选择甘草酸总浓度为1∶10∶30∶60∶120∶240 μmol/L,18α-Gly与18β-Gly物质的量比分别为10∶0、8∶2、6∶4、5∶5、4∶6、2∶8、0∶10,根据细胞存活率,选择合适浓度比例,利用流式细胞仪测定细胞内荧光强度,寻找荧光强度最大的浓度比,即对P-gp抑制作用最强,P-gp功能最弱.研究结果表明甘草酸总浓度为1 μmol/L时,随着18α-Gly比例的减少,荧光强度逐渐减弱,P-gp诱导作用逐渐增加.甘草酸总浓度为10 μmol/L和60 μmol/L时,随着两者物质的量比的变化,荧光强度变化明显;当总浓度为10 μmol/L,n(18α-Gly)∶n(18β-Gly)=4∶6时,荧光强度较强,抑制作用明显;当总浓度为60 μmol/L,两者物质的量比为5∶5时荧光强度最强,抑制作用最强.关键词:18α-甘草酸;18β-甘草酸;P-糖蛋白功能;Caco-2细胞;不同配比/浓度中图分类号:R285.5 文献标志码:A 文章编号:1000-1565(2016)03-0249-08Different proportion and concentration of Glycyrrhizicacid -18-epimers effect on P-gp function in Caco-2 cellsZHAO Yanyan^1,2,SUN Dong¹,LIU Liyan³,LIU Yafei¹,WANG Jing¹(1.College of Chemistry and Environmental Science,Hebei University,Baoding 071002,China;2.College of Pharmaceutical Sciences,Hebei University,Baoding 071002,China;3.Experimental Center of Medicine,Hebei University,Baoding 071000,China)Abstract:To evaluate the effect of glycyrrhizic acid 18 epimers,18α-glycyrrhizic acid(18α-Gly)and 18β-glycyrrhizic acid(18β-Gly)at different proportion and concentration on the P-glycoprotein(P-gp)activity in colon cancer Caco-2 cell to select their appropriate concentration and proportion in use.Making a cell model,selecting the total glycyrrhizic acid concentration of 1,10,30,60,120,240 μmol/L and proportion of 10∶0,8∶2,6∶4,5∶5,4∶6,2∶8,0∶10,determining their optimum concentration and proportion by cell survival rate,and detecting the maximum fluorescence value by flow cytometer,that is,thestrongest inhibitory effect of P-gp and its weakest function.When the concentration of glycyrrhizic acid was 1 μmol/L,the fluorescence intensity became gradually weak and its induction fuction was strengthed gradually with the reduction in proportion of 18α-Gly.When the concentration of glycyrrhizic acid was 10 μmol/L and- 收稿日期:2015-10-08 基金项目:河北省自然科学基金资助项目(H2013201203);河北大学医学学科专项资金建设项目(2014A1003) 第一作者:赵燕燕(1960—),女,天津人,河北大学教授,主要从事药学以及将现代分离技术用于临床、药学、食品、环境等方面的研究.E-mail:zhaoyany606@tom.com第3期赵燕燕等:甘草酸18位差向异构体不同配比和浓度对Caco-2细胞P-gp功能的影响60 μmol/L,the fluorescence intensity changed significantly.When the concentration of glycyrrhizic acid was 10 μmol/L and their proportion was n(18α-Gly)∶n(18β-Gly)=4∶6,the fluorescence intensity became stronger and its inhibitory effect was remarkable.When the concentration of glycyrrhizic acid was 60 μmol/L and their proportion was n(18α-Gly)∶n(18β-Gly)=5∶5,the fluorescence intensity was the highest,and the strongest inhibitory effect of P-gp appeared.

Key words: 18α-glycyrrhizic acid, 18β-glycyrrhizic acid, P-glycoprotein, Caco-2 cell, different proportion/ concentration

中图分类号:

R285.5

赵燕燕,孙东,刘丽艳,柳亚飞,王静. 甘草酸18位差向异构体不同配比和浓度对Caco-2细胞P-gp功能的影响[J]. 河北大学学报(自然科学版), 2016, 36(3): 249-256.

ZHAO Yanyan,SUN Dong,LIU Liyan,LIU Yafei,WANG Jing. Different proportion and concentration of Glycyrrhizic acid -18-epimers effect on P-gp function in Caco-2 cells[J]. Journal of Hebei University (Natural Science Edition), 2016, 36(3): 249-256.

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