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Cited by 77Barrier Structure and Function StudiesDrug Transport and Resistance MechanismsAcute Ischemic Stroke Management

Impact of cigarette smoke extract and hyperglycemic conditions on blood–brain barrier endothelial cells

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Prasad, Shikha, Sajja, Ravi K, Park, Jee Hyun, Naik, Pooja, Kaisar, Mohammad Abul, Cucullo, Luca·2015·Fluids and Barriers of the CNS
DOI 10.1186/s12987-015-0014-xPMC PMC4513397
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Abstract

BACKGROUND: Diabetes and tobacco smoking are significant public health concerns which have been shown to independently impact the blood-brain barrier (BBB). Since smoking is a risk factor for diabetes and shares some of the common pathological pathways leading to metabolic abnormalities, it is hypothesized that their combination would produce additive or synergistic BBB dysfunction. Therefore, the objective of this study was to assess this hypothesis and evaluate the magnitude of these effects in vitro using hCMEC/D3 cells; a well-established human BBB endothelial cell line. METHODS: Monolayers of hCMEC/D3 cells were exposed to hyperglycemic conditions (HG; 35 mM) or 5% soluble cigarette smoke extracts (CSE, model of mainstream smoke exposure) for 12-24 h. Cells were then harvested for subsequent biochemical analyses. Transendothelial electrical resistance (TEER) and paracellular permeability to florescent dextrans were used to assess monolayer integrity. Analysis of released factors and cytokines was carried out by ELISA. Western blot (WB) analysis/immunofluorescence of relevant molecular targets was carried out. P-gp efflux activity was measured using rhodamine 123. RESULTS: Immunofluorescence and WB data showed a significant ZO-1 down-regulation by HG and/or CSE over 24 h exposure. CSE in presence of HG produced a synergistic increase in release of vascular endothelial growth factor that was accompanied by decreased TEER and augmented permeability to labeled dextrans in a size-dependent manner. Moreover, CSE increased the expression of GLUT-1 and SGLT-1 in isolated membrane fractions of hCMEC/D3 cells. The effect was amplified by HG. Both, HG and CSE elicited the membrane upregulation of P-glycoprotein (P-gp) expression which however, was not paralleled by a comparable efflux activity. Interestingly, concomitant exposure to HG and CSE evoked a marked upregulation of PECAM-1 and other pro-inflammatory markers including IL-6 and -8, when compared to each condition alone. Moreover, exposure to all tested conditions amplified (to a different degree) cellular oxidative stress response denoted by increased Nrf2 nuclear translocation. CONCLUSION: Overall, our results have clearly shown an additive pattern in the release of angiogenic and inflammatory factors following concomitant exposure to HG and CSE. This suggests the involvement of common key modulators in BBB impairment by both CS and HG possibly through the activation of oxidative stress responses.

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Authors
Prasad, S., Sajja, R. K., Park, J. H., Naik, P., Kaisar, M. A., & Cucullo, L.
Affiliations
Texas Tech University Health Sciences Center (US)
Corresponding author
Cucullo, Luca
ORCID
Venue
Fluids and Barriers of the CNS
Publisher
Springer Science and Business Media LLC
Published
2015-07-24
Volume
12
Issue
1
Pages
18-18
ISSN
2045-8118 (ISSN-L)
DOI
10.1186/s12987-015-0014-x
PMC
PMC4513397
OpenAlex
W1902782356
Cited by
77
References
61
Citations by year
2026: 7 · 2025: 6 · 2024: 9 · 2023: 3 · 2022: 10 · 2021: 5 · 2020: 7 · 2019: 6 · 2018: 7 · 2017: 6 · 2016: 7 · 2015: 4
FWCI
4.28
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Fluids and Barriers of the CNS (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • Institut National de la Santé et de la Recherche Médicale
  • National Institutes of Health
  • Weill Cornell Medical College
  • National Institute on Drug Abuse (R01DA029121, R01-DA029121-01A1)
Topics
  • Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
  • Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
  • Health Sciences › Medicine › Epidemiology › Acute Ischemic Stroke Management
Keywords
cigarette smoke extract, hyperglycemia, blood-brain barrier, endothelial dysfunction, diabetes mellitus, smoking, ZO-1 tight junction, vascular endothelial growth factor, GLUT1, SGLT1, P-glycoprotein, CD31, inflammatory cytokines, oxidative stress, Nrf2, endothelial permeability, synergistic effect
MeSH terms
Blood-Brain Barrier, Cell Line, Cell Survival, Humans, Hyperglycemia, Inflammation, ATP Binding Cassette Transporter, Subfamily B, Member 1, Endothelial Cells, Glucose Transport Proteins, Facilitative, Tobacco Products, Zonula Occludens-1 Protein

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