CoChat
Sign Up
Cited by 289Barrier Structure and Function StudiesNeuroinflammation and Neurodegeneration MechanismsDrug Transport and Resistance Mechanisms

The DOI matches its Crossref record; 48 of the 49 references it cites were found in a registry or an index.

Interleukin-1β Induces Blood–Brain Barrier Disruption by Downregulating Sonic Hedgehog in Astrocytes

Open access
Wang, Yue, Jin, Shijie, Sonobe, Yoshifumi, Cheng, Yi, Horiuchi, Hiroshi, Parajuli, Bijay, Kawanokuchi, Jun, Mizuno, Tetsuya, Takeuchi, Hideyuki, Suzumura, Akio·2014·PLoS ONE
DOI 10.1371/journal.pone.0110024PMC PMC4196962
Open this paper in CoChat — free accountOpen PDF

Abstract

The blood-brain barrier (BBB) is composed of capillary endothelial cells, pericytes, and perivascular astrocytes, which regulate central nervous system homeostasis. Sonic hedgehog (SHH) released from astrocytes plays an important role in the maintenance of BBB integrity. BBB disruption and microglial activation are common pathological features of various neurologic diseases such as multiple sclerosis, Parkinson's disease, amyotrophic lateral sclerosis, and Alzheimer's disease. Interleukin-1β (IL-1β), a major pro-inflammatory cytokine released from activated microglia, increases BBB permeability. Here we show that IL-1β abolishes the protective effect of astrocytes on BBB integrity by suppressing astrocytic SHH production. Astrocyte conditioned media, SHH, or SHH signal agonist strengthened BBB integrity by upregulating tight junction proteins, whereas SHH signal inhibitor abrogated these effects. Moreover, IL-1β increased astrocytic production of pro-inflammatory chemokines such as CCL2, CCL20, and CXCL2, which induce immune cell migration and exacerbate BBB disruption and neuroinflammation. Our findings suggest that astrocytic SHH is a potential therapeutic target that could be used to restore disrupted BBB in patients with neurologic diseases.

Overview

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.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1371/journal.pone.0110024

Cites

48 of 48 references verified0 citation errors · 0 not checkable

Statistics

Figures

Figure 1 IL-1β abolishes the protective effect of astrocytes on BBB integrity. MEBC4 cells were treated for 24 h with 2 ng/ml IL-1β, ACM, or IL-1β–treated ACM for 24 h. FITC-BSA was loaded onto the luminal side of the insert for 1 h, and then the FITC-BSA levels on the abluminal side were analyzed. All quantitative data are expressed as means ± SEM (n = 5), normalized to the corresponding values from untreated cells. *, p<0.001.
Figure 1 IL-1β abolishes the protective effect of astrocytes on BBB integrity. MEBC4 cells were treated for 24 h with 2 ng/ml IL-1β, ACM, or IL-1β–treated ACM for 24 h. FITC-BSA was loaded onto the luminal side of the insert for 1 h, and then the FITC-BSA levels on the abluminal side were analyzed. All quantitative data are expressed as means ± SEM (n = 5), normalized to the corresponding values from untreated cells. *, p<0.001.CC BY · PubMed Central
Figure 2 IL-1β downregulates SHH production in astrocytes. (A) Shh mRNA levels in astrocytes, determined by qPCR. Astrocytes were treated with IL-1β for 6 h. Values are means ± SEM (n = 5). *, p<0.05; †, p<0.01. (B) Protein levels of SHH in ACM, determined using ELISA. Astrocytes were treated with IL-1β for 24 h. Values are means ± SEM (n = 5). *, p<0.001.
Figure 2 IL-1β downregulates SHH production in astrocytes. (A) Shh mRNA levels in astrocytes, determined by qPCR. Astrocytes were treated with IL-1β for 6 h. Values are means ± SEM (n = 5). *, p<0.05; †, p<0.01. (B) Protein levels of SHH in ACM, determined using ELISA. Astrocytes were treated with IL-1β for 24 h. Values are means ± SEM (n = 5). *, p<0.001.CC BY · PubMed Central
Figure 3 SHH signaling is critical for maintenance of BBB integrity. (A) MBEC4 cells were treated with SHH or the Smo agonist purmorphamine for 24 h. (B) MBEC4 cells were treated with ACM or the Smo antagonist cyclopamine for 24 h. FITC-BSA was loaded onto the luminal side of the insert for 1 h, and then the FITC-BSA levels on the abluminal side were analyzed. All quantitative data are expressed as means ± SEM (n = 5), normalized to the corresponding values from untreated cells. *, p<0.001.
Figure 3 SHH signaling is critical for maintenance of BBB integrity. (A) MBEC4 cells were treated with SHH or the Smo agonist purmorphamine for 24 h. (B) MBEC4 cells were treated with ACM or the Smo antagonist cyclopamine for 24 h. FITC-BSA was loaded onto the luminal side of the insert for 1 h, and then the FITC-BSA levels on the abluminal side were analyzed. All quantitative data are expressed as means ± SEM (n = 5), normalized to the corresponding values from untreated cells. *, p<0.001.CC BY · PubMed Central
Figure 4 Astrocytic SHH signaling regulates expression of tight junction proteins in BBB. Western blotting of claudin-5 (A), occludin (B) and ZO-1 (C) in MBEC4 cells. Cells were treated for 24 h with ACM, SHH (100 ng/ml), the Smo agonist purmorphamine (1 µM), or the Smo antagonist cyclopamine (30 µM). All quantitative data are expressed as means ± SEM (n = 5), normalized to the corresponding values from untreated cells. *, p<0.05.
Figure 4 Astrocytic SHH signaling regulates expression of tight junction proteins in BBB. Western blotting of claudin-5 (A), occludin (B) and ZO-1 (C) in MBEC4 cells. Cells were treated for 24 h with ACM, SHH (100 ng/ml), the Smo agonist purmorphamine (1 µM), or the Smo antagonist cyclopamine (30 µM). All quantitative data are expressed as means ± SEM (n = 5), normalized to the corresponding values from untreated cells. *, p<0.05.CC BY · PubMed Central
Figure 5 IL-1β upregulates production of pro-inflammatory chemokines in astrocytes. (A) Cxcl2, Ccl2, and Ccl20 mRNA levels in astrocytes, determined by qPCR. Astrocytes were treated with IL-1β for 6 h. Values are means ± SEM (n = 5). *, p<0.05; †, p<0.01. (B) Protein levels of CXCL2, CCL2, and CCL20 in astrocytes, determined by ELISA. Astrocytes were treated with IL-1β for 24 h. Values are means ± SEM (n = 5). *, p<0.001.
Figure 5 IL-1β upregulates production of pro-inflammatory chemokines in astrocytes. (A) Cxcl2, Ccl2, and Ccl20 mRNA levels in astrocytes, determined by qPCR. Astrocytes were treated with IL-1β for 6 h. Values are means ± SEM (n = 5). *, p<0.05; †, p<0.01. (B) Protein levels of CXCL2, CCL2, and CCL20 in astrocytes, determined by ELISA. Astrocytes were treated with IL-1β for 24 h. Values are means ± SEM (n = 5). *, p<0.001.CC BY · PubMed Central
Figure 6 Model of the roles of the SHH and IL-1β pathways in the BBB. (A) Model of SHH signaling pathway in brain capillary endothelial cells. Secreted SHH binds and inactivates its receptor Patched-1, which allowed Smoothened to activate the transcription factor Gli-1. Gli-1 upregulates tight junction proteins and enhances BBB integrity. (B) Model of BBB breakdown by IL-1β. Under healthy conditions (left), astrocytes secrete SHH to upregulate tight junction proteins in endothelial cells and maintain BBB integrity. Under pathologic conditions (right), pathogenic stimuli such as amyloid β, DAMPs, or cytokines induce microglia to release IL-1β. IL-1β suppresses astrocytic SHH production, leading to downregulation of tight junction proteins in endothelial cells and disintegrity of the BBB. IL-1β also activates astrocytes to release pro-inflammatory chemokines such as CXCL2, CCL2, and CCL20. These chemokines induce migration of immune cells, thereby worsening BBB disruption and neuroinflammation. Neut, neutrophils; Mo, monocytes; MΦ, macrophage; DC, dendritic cells; T, T cells.
Figure 6 Model of the roles of the SHH and IL-1β pathways in the BBB. (A) Model of SHH signaling pathway in brain capillary endothelial cells. Secreted SHH binds and inactivates its receptor Patched-1, which allowed Smoothened to activate the transcription factor Gli-1. Gli-1 upregulates tight junction proteins and enhances BBB integrity. (B) Model of BBB breakdown by IL-1β. Under healthy conditions (left), astrocytes secrete SHH to upregulate tight junction proteins in endothelial cells and maintain BBB integrity. Under pathologic conditions (right), pathogenic stimuli such as amyloid β, DAMPs, or cytokines induce microglia to release IL-1β. IL-1β suppresses astrocytic SHH production, leading to downregulation of tight junction proteins in endothelial cells and disintegrity of the BBB. IL-1β also activates astrocytes to release pro-inflammatory chemokines such as CXCL2, CCL2, and CCL20. These chemokines induce migration of immune cells, thereby worsening BBB disruption and neuroinflammation. Neut, neutrophils; Mo, monocytes; MΦ, macrophage; DC, dendritic cells; T, T cells.CC BY · PubMed Central

Verification

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.

A few free checks a day · papers already verified open instantly · results are public

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?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Show all 49 references

Cited by

Sorted by citations

Related

See all

Metadata

Authors
Wang, Y., Jin, S., Sonobe, Y., Cheng, Y., Horiuchi, H., Parajuli, B., Kawanokuchi, J., Mizuno, T., Takeuchi, H., & Suzumura, A.
Affiliations
Research Institute of Environmental Medicine, Nagoya University (JP)
Corresponding author
Takeuchi, Hideyuki
ORCID
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2014-10-14
Volume
9
Issue
10
Pages
e110024
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0110024
PMC
PMC4196962
OpenAlex
W1974655099
Cited by
286
References
49
Citations by year
2026: 13 · 2025: 26 · 2024: 28 · 2023: 28 · 2022: 31 · 2021: 38 · 2020: 22 · 2019: 19 · 2018: 24 · 2017: 20 · 2016: 16 · 2015: 19
FWCI
9.55
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Nagoya University Repository (NAGOYA Repository) (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Institutional Repositories DataBase (IRDB) (submittedVersion, cc-by-nc-nd)
Funders
  • Ministry of Education, Culture, Sports, Science and Technology
  • Ministry of Health, Labour and Welfare
  • National Institute of Biomedical Innovation
Topics
  • Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
Keywords
IL-1β, blood-brain barrier disruption, Sonic hedgehog, astrocytes, tight junction proteins, blood-brain barrier permeability, neuroinflammation, chemokines, MCP-1, CCL20, CXCL2, microglial activation, neurological disorders
MeSH terms
Animals, Astrocytes, Blood-Brain Barrier, Cell Line, Down-Regulation, Gene Expression, Chemokines, Tight Junctions, Paracrine Communication, Mice, Interleukin-1beta, Hedgehog Proteins

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