Abstract
Aims It is a huge challenge to understand the blood–brain barrier (BBB), which is a key element in neuroinflammation associated with many brain diseases. The BBB also regulates the passage of xenobiotics into the central nervous system (CNS), and therefore influences drug efficacy. This may be due to the presence of ATP binding cassette transporters such as P-glycoprotein (Pgp) on the BBB, which are efflux pumps known to transport many drugs. The peptide endothelin 1 (ET-1) is involved in different kinds of CNS diseases and neuroinflammation, and is known to modulate Pgp transport activity. Although there are data from animal models, data from human models are scarce. We evaluated Pgp expression and transport activity in adult human brain microvascular endothelial cells (HBMECs) when exposing an adult human in vitro BBB model to ET-1. Methods Adult HBMECs were cocultured with human adult glial cells on a TranswellsR to mimic blood and CNS compartments. These human in vitro BBBs were exposed for 24 h to 100 nM and 10 nM ET-1. Pgp expression was assessed by flow cytometry and its transport activity by measuring radiolabelled digoxin passage. Results After exposure to ET-1, flow cytometry showed no shift of fluorescence intensity for a Pgp specific antibody. The passage of digoxin increased with a significant decrease of Q ratio for 10 nM ET-1. Conclusion Our results show that ET-1 has no effect on Pgp expression of adult HBMECs, but does modulate Pgp transport activity.
References
Arai H, Hori S, Aramori I, Ohkubo H, Nakanishi S (1990) Cloning and expression of a cDNA encoding an endothelin receptor. Nature 348:730–732
Bagnato A, Rosano L, Spinella F, Di Castro V, Tecce R, Natali PG (2004) Endothelin B receptor blockade inhibits dynamics of cell interactions and communications in melanoma cell progression. Cancer Res 64:1436–1443
Ballabh P, Braun A, Nedergaard M (2004) The blood-brain barrier: an overview: structure, regulation, and clinical implications. Neurobiol Dis 16:1–13
Barone FC, Willette RN, Yue TL, Feurestein G (1995) Therapeutic effects of endothelin receptor antagonists in stroke. Neurol Res 17:259–264
Bauer B, Hartz AM, Miller DS (2007) Tumor necrosis factor alpha and endothelin-1 increase P-glycoprotein expression and transport activity at the blood-brain barrier. Mol Pharmacol 71:667–675
Chauhan A, Hahn S, Gartner S, Pardo CA, Netesan SK, McArthur J, Nath A (2007) Molecular programming of endothelin-1 in HIV-infected brain: role of Tat in up-regulation of ET-1 and its inhibition by statins. FASEB J 21:777–789
Cirrito JR, Deane R, Fagan AM, Spinner ML, Parsadanian M, Finn MB, Jiang H, Prior JL, Sagare A, Bales KR, Paul SM, Zlokovic BV, Piwnica-Worms D, Holtzman DM (2005) P-glycoprotein deficiency at the blood-brain barrier increases amyloid-beta deposition in an Alzheimer disease mouse model. J Clin Invest 115:3285–3290
Cordon-Cardo C, O’Brien JP, Casals D, Rittman-Grauer L, Biedler JL, Melamed MR, Bertino JR (1989) Multidrug-resistance gene (P-glycoprotein) is expressed by endothelial cells at blood-brain barrier sites. Proc Natl Acad Sci USA 86:695–698
de Vries HE, Kuiper J, de Boer AG, Van Berkel TJ, Breimer DD (1997) The blood-brain barrier in neuroinflammatory diseases. Pharmacol Rev 49:143–155
Didier N, Banks WA, Creminon C, Reuddre-Bosquet N, Mabondzo A (2002) HIV-1-induced production of endothelin-1 in an in vitro model of the human blood-brain barrier. Neuroreport 13:1179–1183
Didier N, Romero IA, Creminon C, Wijkhuisen A, Grassi J, Mabondzo A (2003) Secretion of interleukin-1beta by astrocytes mediates endothelin-1 and tumour necrosis factor-alpha effects on human brain microvascular endothelial cell permeability. J Neurochem 86:246–254
Ehrenreich H, Costa T, Clouse KA, Pluta RM, Ogino Y, Coligan JE, Burd PR (1993) Thrombin is a regulator of astrocytic endothelin-1. Brain Res 600:201–207
Fernandez C, Buyse M, German-Fattal M, Gimenez F (2004) Influence of the pro-inflammatory cytokines on P-glycoprotein expression and functionality. J Pharm Pharm Sci 7:359–371
Goralski KB, Hartmann G, Piquette-Miller M, Renton KW (2003) Downregulation of mdr1a expression in the brain and liver during CNS inflammation alters the in vivo disposition of digoxin. Br J Pharmacol 139:35–48
Hagenbuch B, Gao B, Meier PJ (2002) Transport of xenobiotics across the blood-brain barrier. News Physiol Sci 17:231–234
Hartz AM, Bauer B, Fricker G, Miller DS (2004) Rapid regulation of P-glycoprotein at the blood-brain barrier by endothelin-1. Mol Pharmacol 66:387–394
Hawkins RA, O’Kane RL, Simpson IA, Vina JR (2006) Structure of the blood-brain barrier and its role in the transport of amino acids. J Nutr 136:218S–226S
Hebert VY, Crenshaw BL, Romanoff RL, Ekshyyan VP, Dugas TR (2004) Effects of HIV drug combinations on endothelin-1 and vascular cell proliferation. Cardiovasc Toxicol 4:117–131
Jiang B, Hebert VY, Zavecz JH, Dugas TR (2006) Antiretrovirals induce direct endothelial dysfunction in vivo. J Acquir Immune Defic Syndr 42:391–395
Josserand V, Pelerin H, de Bruin B, Jego B, Kuhnast B, Hinnen F, Duconge F, Boisgard R, Beuvon F, Chassoux F, umas-Duport C, Ezan E, Dolle F, Mabondzo A, Tavitian B (2006) Evaluation of drug penetration into the brain: a double study by in vivo imaging with positron emission tomography and using an in vitro model of the human blood-brain barrier. J Pharmacol Exp Ther 316:79–86
Kim JH, Kim JH, Park JA, Lee SW, Kim WJ, Yu YS, Kim KW (2006) Blood-neural barrier: intercellular communication at glio-vascular interface. J Biochem Mol Biol 39:339–345
Kodawara T, Masuda S, Wakasugi H, Uwai Y, Futami T, Saito H, Abe T, Inu K (2002) Organic anion transporter oatp2-mediated interaction between digoxin and amiodarone in the rat liver. Pharm Res 19:738–743
Loscher W, Potschka H (2005a) Drug resistance in brain diseases and the role of drug efflux transporters. Nat Rev Neurosci 6:591–602
Loscher W, Potschka H (2005b) Role of drug efflux transporters in the brain for drug disposition and treatment of brain diseases. Prog Neurobiol 76:22–76
Mangahas CR, la Cruz GV, Friedman-Jimenez G, Jamal S (2005) Endothelin-1 induces CXCL1 and CXCL8 secretion in human melanoma cells. J Invest Dermatol 125:307–311
Megard I, Garrigues A, Orlowski S, Jorajuria S, Clayette P, Ezan E, Mabondzo A (2002) A co-culture-based model of human blood-brain barrier: application to active transport of indinavir and in vivo-in vitro correlation. Brain Res 927:153–167
Mikkaichi T, Suzuki T, Onogawa T, Tanemoto M, Mizutamari H, Okada M, Chaki T, Masuda S, Tokui T, Eto N, Abe M, Satoh F, Unno M, Hishinuma T, Inui K, Ito S, Goto J, Abe T (2004) Isolation and characterization of a digoxin transporter and its rat homologue expressed in the kidney. Proc Natl Acad Sci USA 101:3569–3574
Milane A, Fernandez C, Vautier S, Bensimon G, Meininger V, Farinotti R (2007) Minocycline and riluzole brain disposition: interactions with p-glycoprotein at the blood-brain barrier. J Neurochem 103:164–173
Morgan ET, Goralski KB, Piquette-Miller M, Renton KW, Robertson GR, Chaluvadi MR, Charles KA, Clarke SJ, Kacevska M, Liddle C, Richardson TA, Sharma R, Sinal CJ (2008) Regulation of drug-metabolizing enzymes and transporters in infection, inflammation, and cancer. Drug Metab Dispos 36:205–216
Nakajima M, Morimoto S, Takamoto S, Kitano S, Fukuo K, Onishi T, Ogihara T (1994) Endothelin-1 in cerebrospinal fluid in elderly patients with hypertension and dementia. Hypertension 24:97–100
Nie XJ, Olsson Y (1996) Endothelin peptides in brain diseases. Rev Neurosci 7:177–186
Pardridge WM (2007) Blood-brain barrier delivery. Drug Discov Today 12:54–61
Polli JW, Jarrett JL, Studenberg SD, Humphreys JE, Dennis SW, Brouwer KR, Woolley JL (1999) Role of P-glycoprotein on the CNS disposition of amprenavir (141W94), an HIV protease inhibitor. Pharm Res 16:1206–1212
Porcheray F, Viaud S, Rimaniol AC, Leone C, Samah B, Reuddre-Bosquet N, Dormont D, Gras G (2005) Macrophage activation switching: an asset for the resolution of inflammation. Clin Exp Immunol 142:481–489
Rautio J, Humphreys JE, Webster LO, Balakrishnan A, Keogh JP, Kunta JR, Serabjit-Singh CJ, Polli JW (2006) In vitro p-glycoprotein inhibition assays for assessment of clinical drug interaction potential of new drug candidates: a recommendation for probe substrates. Drug Metab Dispos 34:786–792
Rolinski B, Heigermoser A, Lederer E, Bogner JR, Loch O, Goebel FD (1999) Endothelin-1 is elevated in the cerebrospinal fluid of HIV-infected patients with encephalopathy. Infection 27:244–247
Ronaldson PT, Bendayan R (2006) HIV-1 viral envelope glycoprotein gp120 triggers an inflammatory response in cultured rat astrocytes and regulates the functional expression of P-glycoprotein. Mol Pharmacol 70:1087–1098
Saengkhae C, Salerno M, Ades D, Siove A, Le Moyeca L, Migonney V, Garnier-Suillerot A (2007) Ability of carbazole salts, inhibitors of Alzheimer beta-amyloid fibril formation, to cross cellular membranes. Eur J Pharmacol 559:124–131
Sakurai T, Yanagisawa M, Takuwa Y, Miyazaki H, Kimura S, Goto K, Masaki T (1990) Cloning of a cDNA encoding a non-isopeptide-selective subtype of the endothelin receptor. Nature 348:732–735
Sasaki Y, Takimoto M, Oda K, Fruh T, Takai M, Okada T, Hori S (1997) Endothelin evokes efflux of glutamate in cultures of rat astrocytes. J Neurochem 68:2194–2200
Schinelli S (2006) Pharmacology and physiopathology of the brain endothelin system: an overview. Curr Med Chem 13:627–638
Schwartz M, Butovsky O, Kipnis J (2006) Does inflammation in an autoimmune disease differ from inflammation in neurodegenerative diseases? Possible implications for therapy. J Neuroimmune Pharmacol 1:4–10
Shirakami G, Nakao K, Saito Y, Magaribuchi T, Mukoyama M, Arai H, Hosoda K, Suga S, Mori K, Imura H (1993) Low doses of endothelin-1 inhibit atrial natriuretic peptide secretion. Endocrinology 132:1905–1912
Speciale L, Sarasella M, Ruzzante S, Caputo D, Mancuso R, Calvo MG, Guerini FR, Ferrante P (2000) Endothelin and nitric oxide levels in cerebrospinal fluid of patients with multiple sclerosis. J Neurovirol 6(Suppl 2):S62–S66
Suzuki R, Masaoka H, Hirata Y, Marumo F, Isotani E, Hirakawa K (1992) The role of endothelin-1 in the origin of cerebral vasospasm in patients with aneurysmal subarachnoid hemorrhage. J Neurosurg 77:96–100
Takuwa Y, Yanagisawa M, Takuwa N, Masaki T (1989) Endothelin, its diverse biological activities and mechanisms of action. Prog Growth Factor Res 1:195–206
Tansey MG, Frank-Cannon TC, McCoy MK, Lee JK, Martinez TN, McAlpine FE, Ruhn KA, Tran TA (2008) Neuroinflammation in Parkinson’s disease: is there sufficient evidence for mechanism-based interventional therapy? Front Biosci 13:709–717
Tatsuta T, Naito M, Oh-hara T, Sugawara I, Tsuruo T (1992) Functional involvement of P-glycoprotein in blood-brain barrier. J Biol Chem 267:20383–20391
Theron D, de Barraud LS, Tardivel S, Pelerin H, Demeuse P, Mercier C, Mabondzo A, Farinotti R, Lacour B, Roux F, Gimenez F (2003) Influence of tumor necrosis factor-alpha on the expression and function of P-glycoprotein in an immortalised rat brain capillary endothelial cell line, GPNT. Biochem Pharmacol 66:579–587
Uhr M, Ebinger M, Rosenhagen MC, Grauer MT (2005) The anti-Parkinson drug budipine is exported actively out of the brain by P-glycoprotein in mice. Neurosci Lett 383:73–76
Wright CE, Fozard JR (1988) Regional vasodilation is a prominent feature of the haemodynamic response to endothelin in anaesthetized, spontaneously hypertensive rats. Eur J Pharmacol 155:201–203
Yanagisawa M, Kurihara H, Kimura S, Tomobe Y, Kobayashi M, Mitsui Y, Yazaki Y, Goto K, Masaki T (1988) A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature 332:411–415
Yoshimoto S, Ishizaki Y, Kurihara H, Sasaki T, Yoshizumi M, Yanagisawa M, Yazaki Y, Masaki T, Takakura K, Murota S (1990) Cerebral microvessel endothelium is producing endothelin. Brain Res 508:283–285
Ziv I, Fleminger G, Djaldetti R, Achiron A, Melamed E, Sokolovsky M (1992) Increased plasma endothelin-1 in acute ischemic stroke. Stroke 23:1014–1016
Acknowledgements
We would like to thank the ANRS (Agence Nationale de Recherche sur le SIDA) for financial support. We would also like to thank Gabriel Gras (CEA Fontenay-aux-Roses, France) and Jean-François Heilier (Université Catholique de Louvain, Belgium) for their advice in statistics.
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Hembury, A., Mabondzo, A. Endothelin-1 Reduces P-Glycoprotein Transport Activity in an In Vitro Model of Human Adult Blood–brain Barrier. Cell Mol Neurobiol 28, 915–921 (2008). https://doi.org/10.1007/s10571-008-9277-y
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DOI: https://doi.org/10.1007/s10571-008-9277-y