帕金森病细胞旁路开放对FLZ体外跨血脑屏障模型转运特性的影响

中国药物警戒 ›› 2018, Vol. 15 ›› Issue (1): 6-11.

帕金森病细胞旁路开放对FLZ体外跨血脑屏障模型转运特性的影响

刘潜^1,2, 侯金凤², 张金兰², 张丹², 鲍秀琦², 孙华^2,*

1 北京市神经外科研究所,北京 100050;
2 中国医学科学院药物研究所,北京 100050;

收稿日期:2018-02-12 修回日期:2018-02-12 出版日期:2018-01-20 发布日期:2018-02-12
通讯作者: *孙华女,博士,副研究员·硕导,肝脏生化药理学及肿瘤多药耐药性。E-mail：sunhua@imm.ac.cn
作者简介:刘潜,女,博士,中枢神经系统肿瘤及分子药理研究。
基金资助:
国家自然科学基金（81072697）：新型番荔枝有效成分衍生物FLZ脑内代谢特点的研究; 北京市自然科学基金（7122113）：抗帕金森氏病活性成分FLZ通过血脑屏障机制研究

Effect of Paracellular Permeability Increasing on Transport of Squamosamide Derivative FLZ Across the in vitro BBB Models under Parkinson's Disease Pathological

LIU Qian^1,2, HOU Jin-feng², ZHANG Jin-lan², ZHANG Dan², BAO Xiu-qi², SUN Hua^2,*

1 Beijing Neurosurgical Institute, Capital Medical University, Beijing 100050, China;
2 Department of Pharmacology, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China;

Received:2018-02-12 Revised:2018-02-12 Online:2018-01-20 Published:2018-02-12

摘要/Abstract

摘要： 目的考察生理和帕金森病（Parkinson's disease,PD）病理状态下细胞旁路开放对番荔枝酰胺衍生物FLZ体外跨血脑屏障（blood-brain barrier,BBB）转运特性的影响。方法采用体外原代培养的正常和PD模型大鼠脑微血管内皮细胞（rat cerebral microvessel endothelial cells,rCMECs）,与同种属的C6胶质细胞建立非接触式和接触式体外BBB共培养模型,分别考察转运蛋白中与PD相关且人和大鼠BBB均高表达的P-糖蛋白（P-glycoprotein,P-gp）和PD病理下细胞旁路开放对FLZ跨生理和PD病理BBB转运的影响。结果 FLZ在细胞旁路未开放的生理和细胞旁路开放的PD病理体外BBB模型中的跨膜转运,都表现出剂量依赖的极性转运特性,Papp (B~A) > Papp (A~B),具有易透过BBB能力和高外排现象,并呈现出良好的剂量依赖关系。P-gp抑制剂5 μM zosuquidar可以显著降低FLZ外排,增加转运。而细胞旁路途径开放会促进FLZ进入BBB,但外排率与未开放细胞旁路途径无显著差异。表明虽然PD病理状态下细胞旁路通透性增加引起了FLZ入脑量升高,但这一作用依然被P-gp的外排所抵消。结论生理和PD病理状态下影响FLZ透过BBB的主要因素还是由BBB中P-gp介导的外排。

关键词: FLZ, 血脑屏障, 帕金森病, 细胞旁路, P-糖蛋白

Abstract: Objective To study the effect of paracellular permeability on the transport characteristics of squamosamide derivative FLZ across the physiological and Parkinson's disease (PD) pathological in vitro blood-brain barrier (BBB) models. Methods In vitro BBB models mimicking physiological and PD pathological-related BBB properties were constructed by C6 astroglial cells co-cultured with primary normal or PD rat cerebral microvessel endothelial cells (rCMECs) and in vitro permeability experiments of FLZ were carried out. The effects of P-gp inhibitor zosuquidar and increasing paracellular permeability on the in vitro permeability and efflux ratio of FLZ were also investigated. Results The transport of various concentrations of FLZ (1, 5, 10 μM) through the out of contact co-culture model and in contact co-culture model occurred in both A~B and B~A directions, and Papp B~A transport were significantly higher than those for A~B transport at each FLZ concentration. Upon specific blocking of P-gp using zosuquidar increased intracellular accumulation of FLZ both in the physiological and PD pathological in vitro BBB models and resulted in significantly reduced efflux ratio. Results also showed that more brain penetration of FLZ was detected in the PD pathological model compared to the physiological group with increasing paracellular permeability, but the alter of efflux ratio was not observed as P-glycoproteinon mediated FLZ efflux. Conclusion The poor brain penetration of FLZ are mainly due to the P-gp transport system in the BBB under physiological and PD pathological in vitro BBB models.

Key words: FLZ, blood-brain barrier, Parkinson's disease, paracellular, P-glycoprotein

中图分类号:

R965

刘潜, 侯金凤, 张金兰, 张丹, 鲍秀琦, 孙华. 帕金森病细胞旁路开放对FLZ体外跨血脑屏障模型转运特性的影响[J]. 中国药物警戒, 2018, 15(1): 6-11.

LIU Qian, HOU Jin-feng, ZHANG Jin-lan, ZHANG Dan, BAO Xiu-qi, SUN Hua. Effect of Paracellular Permeability Increasing on Transport of Squamosamide Derivative FLZ Across the in vitro BBB Models under Parkinson's Disease Pathological[J]. Chinese Journal of Pharmacovigilance, 2018, 15(1): 6-11.

参考文献

[1] Cardoso F L, Brites D, Brito M A.Looking at the blood-brain barrier: molecular anatomy and possible investigation approaches[J]. Brain research reviews, 2010,64(2):328-363.
[2] Liddelow S A.Fluids and barriers of the CNS: a historical viewpoint[J]. Fluids and barriers of the CNS, 2011,8(1):2.
[3] Ballabh P, Braun A, Nedergaard M.The blood-brain barrier: an overview: structure, regulation, and clinical implications[J]. Neurobiol Dis,2004,16(1):1-13.
[4] Abbott N J, R nnb ck L, Hansson E. Astrocyte-endothelial interactions at the blood-brain barrier[J]. Nature Reviews Neuroscience,2006,7(1):41-53.
[5] Cecchelli R, Berezowski V, Lundquist S, et al.Modelling of the blood-brain barrier in drug discovery and development[J]. Nature Reviews Drug Discovery, 2007,6(8):650-661.
[6] Golden P L, Pollack G M.Blood-brain barrier efflux transport[J]. J Pharm Sci, 2003,92(9):1739-1753.
[7] ElAli A, Hermann D M. ATP-Binding Cassette Transporters and Their Roles in Protecting the Brain[J]. The Neuroscientist, 2011,17(4):423-436.
[8] Liddelow S A.Fluids and barriers of the CNS: a historical viewpoint[J]. Fluids and barriers of the CNS, 2011,8(1):2.
[9] Colgan O C, Collins N T, Ferguson G, et al.Influence of basolateral condition on the regulation of brain microvascular endothelial tight junction properties and barrier function[J]. Brain Res, 2008,1193:84-92.
[10] Etame A B, Diaz R J, Smith C A, et al.Focused ultrasound disruption of the blood-brain barrier: a new frontier for therapeutic delivery in molecular neurooncology[J]. Neurosurgical Focus, 2012,32(1):E3.
[11] Pardridge W M.Tyrosine hydroxylase replacement in experimental Parkinson's disease with transvascular gene therapy[J]. NeuroRx, 2005,2(1):129-138.
[12] Correale J, Villa A.Cellular elements of the blood-brain barrier[J]. Neurochemical research, 2009,34(12):2067-2077.
[13] Liu Q, Hou J, Chen X, et al.P-glycoprotein mediated efflux limits the transport of the novel anti-Parkinson's disease candidate drug FLZ across the physiological and PD pathological in vitro BBB models[J]. PLoS One, 2014,18,9(7):e102442.
[14] Rosenberg G A.Neurological diseases in relation to the blood-brain barrier[J]. Journal of Cerebral Blood Flow & Metabolism, 2012,32(7):1139-1151.
[15] Garbayo E, Ansorena E, Blanco-Prieto M J. Drug development in Parkinson's disease: from emerging molecules to innovative drug delivery systems. Drug development in Parkinson's disease: from emerging molecules to innovative drug delivery systems[J].Maturitas,2013,76(3):272-278.
[16] Lima L O, Scianni A, Rodrigues-de-Paula F. Progressive resistance exercise improves strength and physical performance in people with mild to moderate Parkinson's disease: a systematic review[J]. Journal of physiotherapy, 2013,59(1):7-13.
[17] Zhang D, Zhang J J, Liu G T.The novel squamosamide derivative FLZ protects against 6-hydroxydopamine-induced apoptosis through inhibition of related signal transduction in SH-SY5Y cells[J]. Eur J Pharmacol, 2007,30(561):1-6.
[18] Kanwar J R, Sriramoju B, Kanwar R K.Neurological disorders and therapeutics targeted to surmount the blood-brain barrier[J]. International journal of nanomedicine, 2012,7(7):3259-3278.
[19] Nakagawa S, Deli M A, Kawaguchi H, et al.A new blood-brain barrier model using primary rat brain endothelial cells, pericytes and astrocytes[J].Neurochemistry international, 2009,54(3-4):253-263.
[20] Cecchelli R, Berezowski V, Lundquist S, et al.Modelling of the blood-brain barrier in drug discovery and development[J].Nature Reviews Drug Discovery, 2007,6(8):650-61.
[21] Colgan O C, Collins N T, Ferguson G, et al.Influence of basolateral condition on the regulation of brain microvascular endothelial tight junction properties and barrier function[J].Brain Res , 2008,1193(5):84-92.
[22] 刘潜, 陈晓光, 侯金凤, 等. P-糖蛋白对番荔枝酰胺衍生物FLZ在Caco-2细胞模型中跨膜转运的影响[J].中国药物警戒. 2015,12(2):69-72.
[23] Reichel A, Begley D J, Abbott N J.An overview of in vitro techniques for blood-brain barrier studies[J]. Methods in molecular medicine 2003,89:307-324.
[24] Mahar Doan K M, Humphreys J E, Webster L O, et al. Passive permeability and P-glycoprotein-mediated efflux differentiate central nervous system (CNS) and non-CNS marketed drugs[J]. J Pharmacol Exp Ther, 2002,303(3):1029-1037.
[25] Abbott N J.Blood-brain barrier structure and function and the challenges for CNS drug delivery[J]. Journal of inherited metabolic disease, 2013,36(3):437-449.