Downregulation of blood-brain barrier phenotype by proinflammatory cytokines involves NADPH oxidase-dependent ROS generation: consequences for interendothelial adherens and tight junctions
Open accessAbstract
BACKGROUND AND OBJECTIVES: Blood-brain barrier (BBB) dysfunction is an integral feature of neurological disorders and involves the action of multiple proinflammatory cytokines on the microvascular endothelial cells lining cerebral capillaries. There is still however, considerable ambiguity throughout the scientific literature regarding the mechanistic role(s) of cytokines in this context, thereby warranting a comprehensive in vitro investigation into how different cytokines may cause dysregulation of adherens and tight junctions leading to BBB permeabilization. METHODS: The present study employs human brain microvascular endothelial cells (HBMvECs) to compare/contrast the effects of TNF-α and IL-6 on BBB characteristics ranging from the expression of interendothelial junction proteins (VE-cadherin, occludin and claudin-5) to endothelial monolayer permeability. The contribution of cytokine-induced NADPH oxidase activation to altered barrier phenotype was also investigated. RESULTS: In response to treatment with either TNF-α or IL-6 (0-100 ng/ml, 0-24 hrs), our studies consistently demonstrated significant dose- and time-dependent decreases in the expression of all interendothelial junction proteins examined, in parallel with dose- and time-dependent increases in ROS generation and HBMvEC permeability. Increased expression and co-association of gp91 and p47, pivotal NADPH oxidase subunits, was also observed in response to either cytokine. Finally, cytokine-dependent effects on junctional protein expression, ROS generation and endothelial permeability could all be attenuated to a comparable extent using a range of antioxidant strategies, which included ROS depleting agents (superoxide dismutase, catalase, N-acetylcysteine, apocynin) and targeted NADPH oxidase blockade (gp91 and p47 siRNA, NSC23766). CONCLUSION: A timely and wide-ranging investigation comparing the permeabilizing actions of TNF-α and IL-6 in HBMvECs is presented, in which we demonstrate how either cytokine can similarly downregulate the expression of interendothelial adherens and tight junction proteins leading to elevation of paracellular permeability. The cytokine-dependent activation of NADPH oxidase leading to ROS generation was also confirmed to be responsible in-part for these events.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Once in the circulation, these substances activate a pro-inflammatory immune response mediated by peripheral T cells and macrophages and compromise the integrity of the BBB.
“Once in the circulation, these substances activate a pro-inflammatory immune response mediated by peripheral T cells and macrophages and compromise the integrity of the BBB (Rochfort et al., 2014).”
From Microbiome–microglia connections via the gut–brain axis (2019) · cites this paper as Rochfort et al., 2014
Activated microglia express ROS-generating enzymes such as NADPH oxidase, which are responsible for degeneration of oligodendrocytes and increased BBB permeability by downregulating expression of VE-cadherin, occludin and claudin-5 proteins in microvascular endothelial cells.
“Activated microglia are known to express ROS-generating enzymes, such as NADPH oxidase, which are responsible for degeneration of oligodendrocytes and increased BBB permeability by downregulating the expression of VE-cadherin, occludin and claudin-5 proteins in microvascular ECs (Fischer et al., 2012; Rochfort et al., 2014).”
From The impact of microglial activation on blood-brain barrier in brain diseases (2014) · cites this paper as Rochfort et al., 2014
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Measurement of protein using bicinchoninic acid
Smith, P. K., Krohn, R. I., Hermanson, G. T., Mallia, A. K., Gartner, F. H., Provenzano, M. D., Fujimoto, E. K., Goeke, N. M., Olson, B. J., & Klenk, D. C. (1985) · Analytical Biochemistry · doi:10.1016/0003-2697(85)90442-7 · Cited by 18,482 · see citing works
Griendling, K. K., Sorescu, D., Lassegue, B. P., & Ushio‐Fukai, M. (2000) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/01.atv.20.10.2175 · Cited by 955 · see citing works
Waßmann, S., Stumpf, M., Strehlow, K., Schmid, A., Schieffer, B., Böhm, M., & Nickenig, G. (2003) · Circulation Research · doi:10.1161/01.res.0000115557.25127.8d · Cited by 469 · see citing works
NADPH Oxidase-Dependent Signaling in Endothelial Cells: Role in Physiology and Pathophysiology
Frey, R. S., Ushio‐Fukai, M., & Malik, A. B. (2008) · Antioxidants and Redox Signaling · doi:10.1089/ars.2008.2220 · Cited by 423 · see citing works
Zou, J., & Crews, F. T. (2005) · Brain Research · doi:10.1016/j.brainres.2004.11.014 · Cited by 410 · see citing works
Occludin: One Protein, Many Forms
Cummins, P. M. (2011) · Molecular and Cellular Biology · doi:10.1128/mcb.06029-11 · Cited by 409 · see citing works
Self-citationAveleira, C. A., Lin, C.-M., Abcouwer, S. F., Ambrósio, A. F., & Antonetti, D. A. (2010) · Diabetes · doi:10.2337/db09-1606 · Cited by 403 · see citing works
IL-6 increases endothelial permeability in vitro
MARUO, N., Morita, I., Shirao, M., & Murota, S. (1992) · Endocrinology · doi:10.1210/endo.131.2.1639018 · Cited by 386 · see citing works
Inflammatory Cytokines in Acute Ischemic Stroke
Tuttolomondo, A., Di Raimondo, D., di Sciacca, R., Pinto, A., & Licata, G. (2008) · Current Pharmaceutical Design · doi:10.2174/138161208786848739 · Cited by 325 · see citing works
Förster, C. Y., Burek, M., Romero, I. A., Weksler, B. B., Couraud, P., & Drenckhahn, D. (2008) · The Journal of Physiology · doi:10.1113/jphysiol.2007.146852 · Cited by 306 · see citing works
Role of NAD(P)H Oxidase in Angiotensin II–Induced JAK/STAT Signaling and Cytokine Induction
Schieffer, B., Luchtefeld, M., Braun, S., Hilfiker, A., Hilfiker‐Kleiner, D., & Drexler, H. (2000) · Circulation Research · doi:10.1161/01.res.87.12.1195 · Cited by 273 · see citing works
Bennett, J. L., Basivireddy, J., Kollar, A., Biron, K. E., Reickmann, P., Jefferies, W. A., & McQuaid, S. (2010) · Journal of Neuroimmunology · doi:10.1016/j.jneuroim.2010.08.011 · Cited by 263 · see citing works
Basuroy, S., Bhattacharya, S., Leffler, C. W., & Parfenova, H. (2009) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.00381.2008 · Cited by 254 · see citing works
Roles of Rho/ROCK and MLCK in TNF‐α‐induced changes in endothelial morphology and permeability
McKenzie, J. A. G., & Ridley, A. J. (2007) · Journal of Cellular Physiology · doi:10.1002/jcp.21114 · Cited by 250 · see citing works
PKCζ Regulates TNF-α–Induced Activation of NADPH Oxidase in Endothelial Cells
Frey, R. S., Rahman, A., Kefer, J., Minshall, R. D., & Malik, A. B. (2002) · Circulation Research · doi:10.1161/01.res.0000017631.28815.8e · Cited by 225 · see citing works
Breaking down the barrier: The effects of HIV-1 on the blood–brain barrier
Strazza, M., Pirrone, V., Wigdahl, B., & Nonnemacher, M. R. (2011) · Brain Research · doi:10.1016/j.brainres.2011.05.015 · Cited by 218 · see citing works
Szeto, A., Nation, D. A., Mendez, A. J., Domínguez‐Bendala, J., Brooks, L., Schneiderman, N., & McCabe, P. M. (2008) · American Journal of Physiology-Endocrinology and Metabolism · doi:10.1152/ajpendo.90718.2008 · Cited by 207 · see citing works
Sharief, M. K., & Thompson, E. J. (1992) · Journal of Neuroimmunology · doi:10.1016/0165-5728(92)90087-2 · Cited by 202 · see citing works
Gp91 <i> <sup>phox</sup> </i> is the heme binding subunit of the superoxide-generating NADPH oxidase
Yu, L., Quinn, M. T., Cross, A. R., & Dinauer, M. C. (1998) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.95.14.7993 · Cited by 202 · see citing works
Nishioku, T., Matsumoto, J., Dohgu, S., Sumi, N., Miyao, K., Takata, F., Shuto, H., Yamauchi, A., & Kataoka, Y. (2010) · Journal of Pharmacological Sciences · doi:10.1254/jphs.09292sc · Cited by 168 · see citing works
Manea, A., Tanase, L. I., Raicu, M., & Simionescu, M. (2009) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/atvbaha.109.193896 · Cited by 159 · see citing works
Wilson, C. M., Gaber, M. W., Sabek, O. M., et al. (2009) · International Journal of Radiation Oncology*Biology*Physics · doi:10.1016/j.ijrobp.2009.02.035 · Cited by 152 · see citing works
Roles of IL-6-gp130 Signaling in Vascular Inflammation
Hou, T., Tieu, B. C., Ray, S., Recinos, A., Cui, R., Tilton, R. G., & Brasier, A. R. (2008) · Current Cardiology Reviews · doi:10.2174/157340308785160570 · Cited by 149 · see citing works
Mark, K. S., & Miller, D. W. (1999) · Life Sciences · doi:10.1016/s0024-3205(99)00139-3 · Cited by 143 · see citing works
Role of Caspases in Cytokine-Induced Barrier Breakdown in Human Brain Endothelial Cells
Lopez‐Ramirez, M. A., Fischer, R., Torres-Badillo, C. C., Davies, H. A., Logan, K. E., Pfizenmaier, K., Male, D. K., Sharrack, B., & Romero, I. A. (2012) · The Journal of Immunology · doi:10.4049/jimmunol.1103460 · Cited by 140 · see citing works
Blood–brain barrier and traumatic brain injury
Alves, J. L. (2013) · Journal of Neuroscience Research · doi:10.1002/jnr.23300 · Cited by 135 · see citing works
Aslam, M., Ahmad, N., Srivastava, R., & Hemmer, B. (2011) · Cytokine · doi:10.1016/j.cyto.2011.10.016 · Cited by 135 · see citing works
Pro-inflammatory cytokine TNF-alpha as a neuroprotective agent in the brain
Figiel, I. (2008) · Acta Neurobiologiae Experimentalis · doi:10.55782/ane-2008-1720 · Cited by 124 · see citing works
Lv, S., Song, H.-L., Zhou, Y., Li, L.-X., Cui, W., Wang, W., & Liu, P. (2010) · Liver International · doi:10.1111/j.1478-3231.2010.02211.x · Cited by 120 · see citing works
Szöcs, K. (2004) · PubMed · Cited by 117 · see citing works
Walsh, T. G., Murphy, R. P., Fitzpatrick, P., Rochfort, K. D., Guinan, A. F., Murphy, A. W., & Cummins, P. M. (2011) · Journal of Cellular Physiology · doi:10.1002/jcp.22655 · Cited by 112 · see citing works
Self-citationBasuroy, S., Bhattacharya, S., Tcheranova, D., Qu, Y., Regan, R. F., Leffler, C. W., & Parfenova, H. (2006) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.00032.2006 · Cited by 112 · see citing works
NAD(P)H oxidase mediates the endothelial barrier dysfunction induced by TNF-α
Gertzberg, N., Neumann, P. H., Rizzo, V., & Johnson, A. (2004) · American Journal of Physiology-Lung Cellular and Molecular Physiology · doi:10.1152/ajplung.00116.2003 · Cited by 98 · see citing works
Stat3 Mediates Interelukin-6 Inhibition of Human Endothelial Nitric-oxide Synthase Expression
Saura, M., Zaragoza, C., Bao, C., Herránz, B., Rodrı́guez-Puyol, M., & Lowenstein, C. J. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m606279200 · Cited by 90 · see citing works
Cohen, S. S., Min, M., Cummings, E. E., Chen, X. F., Sadowska, G. B., Sharma, S., & Stonestreet, B. S. (2013) · NeuroImmunoModulation · doi:10.1159/000350470 · Cited by 70 · see citing works
Role of glutathione redox cycle in TNF-α-mediated endothelial cell dysfunction
Toborek, M. J., Barger, S. W., Mattson, M. P., McClain, C. J., & Hennig, B. (1995) · Atherosclerosis · doi:10.1016/0021-9150(95)05568-h · Cited by 67 · see citing works
Bluyssen, H. A. R., Rastmanesh, M. M., Tilburgs, C., Jie, K. E., Wesseling, S., Goumans, M., de Boer, P., Joles, J. A., & Braam, B. G. (2010) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.00513.2009 · Cited by 66 · see citing works
The effect of TNF-α and IL-6 on the permeability of the rat blood-retinal barrier in vivo
Bamforth, S. D., Lightman, S., & Greenwood, J. R. (1996) · Acta Neuropathologica · doi:10.1007/s004010050476 · Cited by 57 · see citing works
Effects of TNF-α and IFN-γ on Nitric Oxide-Induced Neurotoxicity in the Mouse Brain
Blais, V., & Rivest, S. (2004) · The Journal of Immunology · doi:10.4049/jimmunol.172.11.7043 · Cited by 55 · see citing works
Tang, C., Xue, H., Bai, C., & Fu, R. (2010) · Phytotherapy Research · doi:10.1002/ptr.3278 · Cited by 54 · see citing works
Gloor, S. M., Weber, A., Adachi, N., & Frei, K. (1997) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.1997.7557 · Cited by 52 · see citing works
Zhang, S., Liu, X., GOLDSTEIN, S., et al. (2012) · Molecular Medicine Reports · doi:10.3892/mmr.2012.1159 · Cited by 47 · see citing works
TNF-α and IL-1β Increase Pericyte/Endothelial Cell Co-Culture Permeability
Kerkar, S. P., Williams, M., Blocksom, J. M., Wilson, R. F., Tyburski, J. G., & Steffes, C. P. (2005) · Journal of Surgical Research · doi:10.1016/j.jss.2005.06.033 · Cited by 47 · see citing works
Cytokine-Regulated Protein Degradation by the Ubiquitination System
Baek, K. (2006) · Current Protein and Peptide Science · doi:10.2174/138920306776359740 · Cited by 44 · see citing works
Sury, M. D., Frese‐Schaper, M., Mühlemann, M. K., et al. (2006) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2006.07.016 · Cited by 42 · see citing works
Neuroinflammatory Cytokines - The Common Thread in Alzheimer Pathogenesis
Griffin, W. S. T., Barger, S. W., & Chairman, D. P. A. V. (2011) · European Neurological Review · doi:10.17925/enr.2011.06.02.89 · Cited by 39 · see citing works
Oxidative mechanisms of IL-4-induced IL-6 expression in vascular endothelium
Lee, Y. W., Lee, W. H., & Kim, P. H. (2009) · Cytokine · doi:10.1016/j.cyto.2009.08.009 · Cited by 39 · see citing works
Beling, A., Rahnefeld, A., Kloetzel, P. M., & Krüger, E. (2013) · Frontiers in Physiology · doi:10.3389/fphys.2013.00042 · Cited by 36 · see citing works
Neuroinflammatory Cytokines-The Common Thread in Alzheimer's Pathogenesis.
Griffin, W. S. T., & Barger, S. W. (2010) · PubMed · Cited by 35 · see citing works
Muzaffar, S., Shukla, N., Angelini, G. D., & Jeremy, J. Y. (2006) · European Journal of Pharmacology · doi:10.1016/j.ejphar.2006.03.047 · Cited by 32 · see citing works
Neuroinflammatory Cytokines—The Common Thread in Alzheimer Pathogenesis
Griffin, W. S. T., Barger, S. W., & Chairman, D. P. A. V. (2010) · touchREVIEWS in Neurology · doi:10.17925/usn.2010.06.02.19 · Cited by 32 · see citing works
Breaking the Barrier in Stroke: What Should we Know? A Mini-Review
Borlongan, C. V., Rodrigues, A. A., & Oliveira, M. C. (2012) · Current Pharmaceutical Design · doi:10.2174/138161212802002670 · Cited by 31 · see citing works
Effects of fumaric acid esters on blood–brain barrier tight junction proteins
Bénardais, K., Pul, R., Singh, V., Skripuletz, T., Lee, D.-H., Linker, R. A., Gudi, V., & Stangel, M. (2013) · Neuroscience Letters · doi:10.1016/j.neulet.2013.09.038 · Cited by 24 · see citing works
Guinan, A. F., Rochfort, K. D., Fitzpatrick, P. A., Walsh, T. G., Pierotti, A. R., Phelan, S., Murphy, R. P., & Cummins, P. M. (2013) · Cardiovascular Research · doi:10.1093/cvr/cvt127 · Cited by 18 · see citing works
Self-citationFitzpatrick, P., Guinan, A. F., Walsh, T. G., et al. (2009) · The International Journal of Biochemistry & Cell Biology · doi:10.1016/j.biocel.2009.05.011 · Cited by 16 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
The impact of microglial activation on blood-brain barrier in brain diseases
da Fonseca, A. C. C., Matias, D., Garcia, C., Amaral, R., Geraldo, L. H., Freitas, C., & Lima, F. R. S. (2014) · Frontiers in Cellular Neuroscience · doi:10.3389/fncel.2014.00362 · Cited by 572 · see citing works
Microbiome–microglia connections via the gut–brain axis
Abdel-Haq, R., Schlachetzki, J. C. M., Glass, C. K., & Mazmanian, S. K. (2019) · Journal of Experimental Medicine · doi:10.1084/jem.20180794 · Cited by 471 · see citing works
Focusing on claudin-5: A promising candidate in the regulation of BBB to treat ischemic stroke
Lv, J., Hu, W., Yang, Z., Li, T., Jiang, S., Ma, Z., Chen, F., & Yang, Y. (2018) · Progress in Neurobiology · doi:10.1016/j.pneurobio.2017.12.001 · Cited by 148 · see citing works
Neuroinflammation as an etiological trigger for depression comorbid with inflammatory bowel disease
Craig, C. F., Filippone, R. T., Stavely, R., Bornstein, J. C., Apostolopoulos, V., & Nurgali, K. (2022) · Journal of Neuroinflammation · doi:10.1186/s12974-021-02354-1 · Cited by 124 · see citing works
Brain inflammation and hypertension: the chicken or the egg?
Winklewski, P. J., Radkowski, M., Wszedybyl-Winklewska, M., & Demkow, U. (2015) · Journal of Neuroinflammation · doi:10.1186/s12974-015-0306-8 · Cited by 116 · see citing works
Cain, M. D., Salimi, H., Gong, Y., Yang, L., Hamilton, S. L., Heffernan, J. R., Hou, J., Miller, M. J., & Klein, R. S. (2017) · Journal of Neuroimmunology · doi:10.1016/j.jneuroim.2017.04.008 · Cited by 86 · see citing works
Microbiota‐microglia connections in age‐related cognition decline
Zhou, R., Qian, S., Cho, W. C. S., Zhou, J., Jin, C., Zhong, Y., Wang, J., Zhang, X., Xu, Z., Tian, M., et al. (2022) · Aging Cell · doi:10.1111/acel.13599 · Cited by 71 · see citing works
Clinical Phenotype of Depression Affects Interleukin-6 Synthesis
Zadka, Ł., Dzięgiel, P., Kulus, M., & Olajossy, M. (2017) · Journal of Interferon & Cytokine Research · doi:10.1089/jir.2016.0074 · Cited by 20 · see citing works
- Authors
- Rochfort, K. D., Collins, L. E., Murphy, R. P., & Cummins, P. M.
- Affiliations
- Dublin City University (IE)
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2014-07-03
- Volume
- 9
- Issue
- 7
- Pages
- e101815
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC4081725
- OpenAlex
- W2010985903
- Cited by
- 261
- References
- 55
- Citations by year
- 2026: 21 · 2025: 19 · 2024: 21 · 2023: 25 · 2022: 20 · 2021: 30 · 2020: 25 · 2019: 21 · 2018: 17 · 2017: 21 · 2016: 18 · 2015: 17 · 2014: 5
- FWCI
- 10.59
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- PLoS ONE (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Figshare (submittedVersion, cc-by)
- Funders
- Higher Education Authority (PRTLI Cycle 4)
- Topics
- Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
- Health Sciences › Medicine › Immunology and Allergy › Cell Adhesion Molecules Research
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Keywords
- blood-brain barrier dysfunction, inflammatory cytokines, TNF-α, interleukin-6, NADPH oxidase, reactive oxygen species, adherens junctions, tight junctions, VE-cadherin, occludin, claudin-5, brain microvascular endothelial cells, endothelial permeability, antioxidant therapy, NSC23766
- MeSH terms
- Blood-Brain Barrier, Cells, Cultured, Gene Expression Regulation, Humans, Tumor Necrosis Factor-alpha, Down-Regulation, Interleukin-6, Reactive Oxygen Species, Tight Junctions, NADPH Oxidases, Adherens Junctions, Endothelial Cells
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
