有机阴离子转运体1和3在不同药性中药成分跨膜转运中的作用
|
摘要
目的研究有机阴离子转运体1和3(OAT1、OAT3)在不同药性(苦、辛、酸、甘)中药成分的跨膜转运过程中的作用。方法建立OAT1、OAT3过表达单克隆细胞株(MDCK-OAT1、S2-OAT3),通过100μmol·L~(-1)不同药性成分对OAT1、OAT3介导放射性标记探针底物转运活性的影响,预测各药性成分对OAT1、OAT3的抑制作用;测定不同药性成分对OAT1、OAT3的抑制作用的半数抑制浓度(IC_(50));测定OAT1、OAT3过表达细胞株(MDCK-OAT1、S2-OAT3)和空白对照(MDCK-Mock、S2-Mock)对不同药性中药成分的摄取情况,判断各药性成分是否为OAT1、OAT3的底物,从而预测OAT1、OAT3在不同药性成分跨膜转运中的作用。结果 100μmol·L~(-1)的辛味药和酸味药对OAT1和OAT3有较强的抑制作用,其中川陈皮素、没食子酸、原儿茶酸和β-甘草次酸对OAT1抑制作用的IC_(50)分别是1. 86,2. 88,4. 58和10. 43μmol·L~(-1);欧前胡素、没食子酸、原儿茶酸和β-甘草次酸对OAT3抑制作用的IC_(50)分别是3. 15,3. 39,3. 69和3. 93μmol·L~(-1)。与空白对照相比,OAT1转运体细胞对延胡索乙素、川陈皮素、欧前胡素和异欧前胡素具有较强的摄取能力; OAT3对延胡索乙素、原阿片碱、川陈皮素、欧前胡素和异欧前胡素具有较强的摄取能力。结论多数辛味药为OAT1、OAT3的转运底物,因此OAT1、OAT3对多数辛味药具有较强的摄取能力;而多数的酸味药为OAT1、OAT3的抑制剂,能够抑制OAT1、OAT3对其他底物的转运活性。
Objective To study the role of organic anion transporters 1,3( OAT1 and OAT3) in the transmembrane transport process of Chinese herb ingredients with different properties such as bitter,pungent,sour and sweet. Methods The OAT1-overexpressed cell line( MDCK-OAT1) and the OAT3-overexpressed cell line( S2-OAT3) was constructed. The inhibition of different ingredients to the transport activity of OAT1 and OAT3 were preliminary screened at 100 μmol·L~(-1) concentration. Furthermore,the half maximal inhibitory concentration( IC_(50)) of different medicinal ingredients to transport activity of OAT1 and OAT3 were also determined. In order to predict the ingredients whether or not the substrates of OAT1 and OAT3,the uptake pattern of them were measured in the OAT1-overexpressed cell line( MDCK-OAT1) and the OAT3-overexpressed cell line( S2-OAT3) and the blank control( S2-Mock and MDCK-Mock). Results 100 μmol·L~(-1) pungent-taste herbs and sour-taste herbs showed strong inhibition to OAT1 and OAT3. The IC_(50) of nobiletin,gallic acid,protocatechuic acid,glycyrrhetinic acid on OAT1 were 1. 86,2. 88,4. 58,and 10. 43 μmol·L~(-1) separately. While imperatorin,gallic acid,protocatechuic acid,glycyrrhetinic acid showed strong inhibition to OAT3. The IC_(50) of imperatorin,gallic acid,protocatechuic acid,glycyrrhetinic acid on OAT3 were 3. 15,3. 39,3. 69,and 3. 93 μmol·L~(-1) separately. The results of uptake study showed that tetrahydropalmatine,nobiletin,imperatorin isoimperatorin and hesperidincan can be absorbed by MDCK-OAT1 while the tetrahydropalmatine,protopine,nobiletin,imperatorin,isoimperatorin and hesperidincan can be absorbed by S2-OAT3. Conclusion Most pungent-taste herbs are the substrate of OAT1 and OAT3 which can be identified by OAT1 and OAT3. Most sour-taste herbs herbs are the inhibitors of OAT1 and OAT3,which can inhibit the transport activity of OAT1 and OAT3 to other substrates.
Objective To study the role of organic anion transporters 1,3( OAT1 and OAT3) in the transmembrane transport process of Chinese herb ingredients with different properties such as bitter,pungent,sour and sweet. Methods The OAT1-overexpressed cell line( MDCK-OAT1) and the OAT3-overexpressed cell line( S2-OAT3) was constructed. The inhibition of different ingredients to the transport activity of OAT1 and OAT3 were preliminary screened at 100 μmol·L~(-1) concentration. Furthermore,the half maximal inhibitory concentration( IC_(50)) of different medicinal ingredients to transport activity of OAT1 and OAT3 were also determined. In order to predict the ingredients whether or not the substrates of OAT1 and OAT3,the uptake pattern of them were measured in the OAT1-overexpressed cell line( MDCK-OAT1) and the OAT3-overexpressed cell line( S2-OAT3) and the blank control( S2-Mock and MDCK-Mock). Results 100 μmol·L~(-1) pungent-taste herbs and sour-taste herbs showed strong inhibition to OAT1 and OAT3. The IC_(50) of nobiletin,gallic acid,protocatechuic acid,glycyrrhetinic acid on OAT1 were 1. 86,2. 88,4. 58,and 10. 43 μmol·L~(-1) separately. While imperatorin,gallic acid,protocatechuic acid,glycyrrhetinic acid showed strong inhibition to OAT3. The IC_(50) of imperatorin,gallic acid,protocatechuic acid,glycyrrhetinic acid on OAT3 were 3. 15,3. 39,3. 69,and 3. 93 μmol·L~(-1) separately. The results of uptake study showed that tetrahydropalmatine,nobiletin,imperatorin isoimperatorin and hesperidincan can be absorbed by MDCK-OAT1 while the tetrahydropalmatine,protopine,nobiletin,imperatorin,isoimperatorin and hesperidincan can be absorbed by S2-OAT3. Conclusion Most pungent-taste herbs are the substrate of OAT1 and OAT3 which can be identified by OAT1 and OAT3. Most sour-taste herbs herbs are the inhibitors of OAT1 and OAT3,which can inhibit the transport activity of OAT1 and OAT3 to other substrates.
引文
[1]MANDAL A,AGRAHARI V,KHURANA V,et al. Transporter effects on cell permeability in drug delivery[J]. Exp Opin Drug Delivery,2017,14(3):385-401.
[2]张铁军,许浚,申秀萍,等.基于中药质量标志物(Q-Marker)的元胡止痛滴丸的“性-效-物”三元关系和作用机制研究[J].中草药,2016,47(13):2199-2211.
[3]武卫党,张星艳,魏滋鸿,等.小檗碱对有机阴离子转运体抑制作用及双向跨膜转运研究[J].药物评价研究,2017,49(6):778-782.
[4]谭润雅,雷鹏,周政,等. HPLC法同时测定枸橘中欧前胡素和异欧前胡素的含量[J].中药新药与临床药理,2016,27(2):255-258.
[5]徐欢,陈海芳,介磊,等. HPLC法测定枳壳中川陈皮素,红橘素的含量[J].药物分析杂志,2009,38(9):1411-1414.
[6]黄晓丹. HPLC法同时测定炙甘草汤中甘草苷、甘草素、异甘草素的含量[J].中国药房,2012,23(35):3346-3348.
[7]ZHANG Z Y,LIU C X. LC/MS/MS and radioisotope method combined for recognizing the affinity between catalpol and OCT2 transporter[J]. Front Chem Sci Engineering,2012,6(4):436-442.
[8]UWAI Y,HONJO E. Transport of xanthurenic acid by rat/human organic anion transporters OAT1 and OAT3[J]. J Agricult Chem Society Jpn,2013,77(7):1517-1521.
[2]张铁军,许浚,申秀萍,等.基于中药质量标志物(Q-Marker)的元胡止痛滴丸的“性-效-物”三元关系和作用机制研究[J].中草药,2016,47(13):2199-2211.
[3]武卫党,张星艳,魏滋鸿,等.小檗碱对有机阴离子转运体抑制作用及双向跨膜转运研究[J].药物评价研究,2017,49(6):778-782.
[4]谭润雅,雷鹏,周政,等. HPLC法同时测定枸橘中欧前胡素和异欧前胡素的含量[J].中药新药与临床药理,2016,27(2):255-258.
[5]徐欢,陈海芳,介磊,等. HPLC法测定枳壳中川陈皮素,红橘素的含量[J].药物分析杂志,2009,38(9):1411-1414.
[6]黄晓丹. HPLC法同时测定炙甘草汤中甘草苷、甘草素、异甘草素的含量[J].中国药房,2012,23(35):3346-3348.
[7]ZHANG Z Y,LIU C X. LC/MS/MS and radioisotope method combined for recognizing the affinity between catalpol and OCT2 transporter[J]. Front Chem Sci Engineering,2012,6(4):436-442.
[8]UWAI Y,HONJO E. Transport of xanthurenic acid by rat/human organic anion transporters OAT1 and OAT3[J]. J Agricult Chem Society Jpn,2013,77(7):1517-1521.