马钱子生物碱类成分在MDCK-MDR1单层细胞模型中的转运机制研究
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摘要
目的研究马钱子碱、士的宁在MDCK-MDR1单层细胞模型中的双向转运机制。方法 MTT法确定马钱子碱、士的宁对MDCK-MDR1单层细胞毒性范围,以MDCK-MDR1单层细胞为体外研究模型,考察转运时间、药物作用质量浓度、P-糖蛋白(P-gp)抑制剂维拉帕米对马钱子碱、士的宁的累积吸收浓度(C_(cum))和表观渗透系数(P_(app))的影响。结果马钱子碱和士的宁P_(app)均>1×10~(-5)cm/s,P_(app)(BL→AP)/P_(app)(AP→BL)均<2。马钱子碱、士的宁与维拉帕米合用,显著降低了P_(app)(BL→AP)/P_(app)(AP→BL)值。结论马钱子碱、士的宁以被动转运为主,P-gp抑制剂维拉帕米对其吸收有明显抑制作用,马钱子碱、士的宁可能是P-gp底物。
Objective To study the bi-direction transport behavior of brucine and strychnine in the MDCK-MDR1 cell monolayer model.Methods MTT method was employed to confirm the safe concentration of brucine and strychnine towards MDCK-MDR1 cells. The effects of transport time, drug concentration, and P-glycoprotein inhibitor verapamil on cumulative absorption concentration(C_(cum)) and apparent permeability coefficient(P_(app)) of brucine and strychnine in MDCK-MDR1 monolayer cells were studied. Results The P_(app) value of brucine and strychnine was larger than 1 × 10~(-5) cm/s and the ratio of P_(app)(BL→AP) vs P_(app)(AP→BL) was less than 2. Brucine/strychnine combined with verapamil decreased the ratio of P_(app)(BL→AP) vs P_(app)(AP→BL). Conclusion The absorption of brucine and strychnine in MDCK-MDR1 cell monolayer model was well and the passive transference was its main intestinal absorption mechanism. The P-gp inhibitor verapamil has a significant inhibitory effect on brucine and strychnine absorption. Brucine and strychnine may be a substrate of P-glycoprotein.
Objective To study the bi-direction transport behavior of brucine and strychnine in the MDCK-MDR1 cell monolayer model.Methods MTT method was employed to confirm the safe concentration of brucine and strychnine towards MDCK-MDR1 cells. The effects of transport time, drug concentration, and P-glycoprotein inhibitor verapamil on cumulative absorption concentration(C_(cum)) and apparent permeability coefficient(P_(app)) of brucine and strychnine in MDCK-MDR1 monolayer cells were studied. Results The P_(app) value of brucine and strychnine was larger than 1 × 10~(-5) cm/s and the ratio of P_(app)(BL→AP) vs P_(app)(AP→BL) was less than 2. Brucine/strychnine combined with verapamil decreased the ratio of P_(app)(BL→AP) vs P_(app)(AP→BL). Conclusion The absorption of brucine and strychnine in MDCK-MDR1 cell monolayer model was well and the passive transference was its main intestinal absorption mechanism. The P-gp inhibitor verapamil has a significant inhibitory effect on brucine and strychnine absorption. Brucine and strychnine may be a substrate of P-glycoprotein.
引文
[1]中国药典[S].一部.2015.
[2]Pastan I,Gottesman M M,Ueda K,et al.A retrovirus carrying an MDR1 cDNA confers multidrug resistance and polarized expression of p-glycoprotrin in MDCKcells[J].Proc Natl Acad Sci USA,1998,85(12):4486-4490.
[3]Horio M,Chin K V,Currier S J,et al.Transepithelial transport of drugs by the mulitidrug transporter in cultured Madin-Darby canine kidney cell epithelia[J].JBiol Chem,1989,264(25):14880-14884.
[4]Hong L,Xu C,O’Neal S,et al.Roles of P-glycoprotein and multidrug resistance protein in transporting para-aminosalicylic acid and its N-acetylated metabolite in mice brain[J].Acta Pharmacol Sin,2014,35(12):1577-1585.
[5]Hu P Y,Liu D,Qing Q Z,et al.Elucidation of transport mechanism of paeoniflorin and the influence of ligustilide,senkyunolide I and senkyunolide A on paeoniflorin transport through MDCK-MDR1 cells as blood-brain barrier in vitro model[J].Molecules,2016,doi:10.3390/molecules21030300.
[6]Xu D H,Yan M,Li H D,et al.Influence of P-glycoprotein on brucine transport at the in vitro blood-brain barrier[J].Europ J Pharmacol,2012,690(1/3):68-76.
[7]Ren T K,Li M L,Zheng H,et al.Microdialysis combined with RRLC-MS/MS for the pharmacokinetics of two major alkaloids of Bi qi capsule and the potential roles of P-gp and BCRP on their penetration[J].J Chromatogr B,2018,1092(15):72-81.
[8]Lécuyer M A,Kebir H,Prat A,et al.Glial influences on BBB functions and molecular players in immune cell trafficking[J].Biochim Biophys Acta Mol Basis Disease,2016,1862(3):472-482.
[9]Candela P,Saint-Pol J,Kuntz M,et al.In vitro discrimination of the role of LRP1 at the BBB cellular level:Focus on brain capillary endothelial cells and brain pericytes[J].Brain Res,2015,1594(12):15-26.
[10]朱狄峰.丹参素及芍药苷在Caco-2细胞模型中转运的研究[D].杭州:浙江大学,2006.
[11]杨冰,张紫薇,赵博,等.微乳对葛根素在MDCK-MDR1细胞的转运影响及机制研究[J].中草药,2018,49(12):2890-2896.
[12]刘强,高小玲,柴逸峰,等.四氢帕马丁在MDCK-MDR1细胞系中的跨膜转运机制[J].药学服务与研究,2012,12(1):34-37.
[13]Mariline G,Renata S,Carolina R P,et al.Cellular models and in vitro assays for the screening of modulators of P-gp,MRP1 and BCRP[J].Molecules,2017,doi:10.3390/molecules22040600.
[14]Yang B,Du S Y,et al.Influence of paeoniflorin and menthol on puerarin transport across MDCK and MDCK-MDR1 cells as blood-brain barrier in vitro model[J].J Pharm Pharmacol,2018,70(3):349-360.
[15]王俊俊,廖晓欢,叶敏,等.Caco-2单层细胞模型上士的宁的体外吸收机制及其与甘草苷的转运相互作用[J].药学学报,2010,45(9):1160-1164.
[16]叶敏.马钱子碱、次乌头碱的体外吸收机制及其他们分别与甘草苷的转运相互作用研究[D].武汉:湖北大学,2012.
[17]陈海波,马凤森,方剑乔,等.马钱子组分对人HaCaT细胞毒性作用的比较[J].中成药,2015,37(1):16-21.
[18]Liu L L,Guan Y M,Lu X P,et al.Mechanisms of P-glycoprotein modulation by Semen Strychni combined with Radix Paeoniae Alba[J].Evidence-Based Compl Altern Med,2017,doi:10.1155/2017/1743870.
[19]Li H,Jin H E,Kim W,et al.Involvement of P-glycoprotein,multidrug resistance protein 2 and breast cancer resistance protein in the transport of belotecan and topotecan in Caco-2 and MDCKII cells[J].Res Paper,2008,25(11):2601-2612.
[20]Chen Z Z,Lu Y,Du S Y,et al.Influence of borneol and muscone on geniposide transport through MDCK and MDCK-MDR1 cells as blood-brain barrier in vitro model[J].Int J Pharm,2013,456(1):73-79.
[21]郑琴,周欢,熊文海,等.乌头碱在Caco-2细胞模型上的转运行为[J].中国实验方剂学杂志,2014,20(20):121-124.
[2]Pastan I,Gottesman M M,Ueda K,et al.A retrovirus carrying an MDR1 cDNA confers multidrug resistance and polarized expression of p-glycoprotrin in MDCKcells[J].Proc Natl Acad Sci USA,1998,85(12):4486-4490.
[3]Horio M,Chin K V,Currier S J,et al.Transepithelial transport of drugs by the mulitidrug transporter in cultured Madin-Darby canine kidney cell epithelia[J].JBiol Chem,1989,264(25):14880-14884.
[4]Hong L,Xu C,O’Neal S,et al.Roles of P-glycoprotein and multidrug resistance protein in transporting para-aminosalicylic acid and its N-acetylated metabolite in mice brain[J].Acta Pharmacol Sin,2014,35(12):1577-1585.
[5]Hu P Y,Liu D,Qing Q Z,et al.Elucidation of transport mechanism of paeoniflorin and the influence of ligustilide,senkyunolide I and senkyunolide A on paeoniflorin transport through MDCK-MDR1 cells as blood-brain barrier in vitro model[J].Molecules,2016,doi:10.3390/molecules21030300.
[6]Xu D H,Yan M,Li H D,et al.Influence of P-glycoprotein on brucine transport at the in vitro blood-brain barrier[J].Europ J Pharmacol,2012,690(1/3):68-76.
[7]Ren T K,Li M L,Zheng H,et al.Microdialysis combined with RRLC-MS/MS for the pharmacokinetics of two major alkaloids of Bi qi capsule and the potential roles of P-gp and BCRP on their penetration[J].J Chromatogr B,2018,1092(15):72-81.
[8]Lécuyer M A,Kebir H,Prat A,et al.Glial influences on BBB functions and molecular players in immune cell trafficking[J].Biochim Biophys Acta Mol Basis Disease,2016,1862(3):472-482.
[9]Candela P,Saint-Pol J,Kuntz M,et al.In vitro discrimination of the role of LRP1 at the BBB cellular level:Focus on brain capillary endothelial cells and brain pericytes[J].Brain Res,2015,1594(12):15-26.
[10]朱狄峰.丹参素及芍药苷在Caco-2细胞模型中转运的研究[D].杭州:浙江大学,2006.
[11]杨冰,张紫薇,赵博,等.微乳对葛根素在MDCK-MDR1细胞的转运影响及机制研究[J].中草药,2018,49(12):2890-2896.
[12]刘强,高小玲,柴逸峰,等.四氢帕马丁在MDCK-MDR1细胞系中的跨膜转运机制[J].药学服务与研究,2012,12(1):34-37.
[13]Mariline G,Renata S,Carolina R P,et al.Cellular models and in vitro assays for the screening of modulators of P-gp,MRP1 and BCRP[J].Molecules,2017,doi:10.3390/molecules22040600.
[14]Yang B,Du S Y,et al.Influence of paeoniflorin and menthol on puerarin transport across MDCK and MDCK-MDR1 cells as blood-brain barrier in vitro model[J].J Pharm Pharmacol,2018,70(3):349-360.
[15]王俊俊,廖晓欢,叶敏,等.Caco-2单层细胞模型上士的宁的体外吸收机制及其与甘草苷的转运相互作用[J].药学学报,2010,45(9):1160-1164.
[16]叶敏.马钱子碱、次乌头碱的体外吸收机制及其他们分别与甘草苷的转运相互作用研究[D].武汉:湖北大学,2012.
[17]陈海波,马凤森,方剑乔,等.马钱子组分对人HaCaT细胞毒性作用的比较[J].中成药,2015,37(1):16-21.
[18]Liu L L,Guan Y M,Lu X P,et al.Mechanisms of P-glycoprotein modulation by Semen Strychni combined with Radix Paeoniae Alba[J].Evidence-Based Compl Altern Med,2017,doi:10.1155/2017/1743870.
[19]Li H,Jin H E,Kim W,et al.Involvement of P-glycoprotein,multidrug resistance protein 2 and breast cancer resistance protein in the transport of belotecan and topotecan in Caco-2 and MDCKII cells[J].Res Paper,2008,25(11):2601-2612.
[20]Chen Z Z,Lu Y,Du S Y,et al.Influence of borneol and muscone on geniposide transport through MDCK and MDCK-MDR1 cells as blood-brain barrier in vitro model[J].Int J Pharm,2013,456(1):73-79.
[21]郑琴,周欢,熊文海,等.乌头碱在Caco-2细胞模型上的转运行为[J].中国实验方剂学杂志,2014,20(20):121-124.