Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Mar 21;16(7):5905-5915.
doi: 10.18632/aging.205678. Epub 2024 Mar 21.

Cepharanthine maintains integrity of the blood-brain barrier (BBB) in stroke via the VEGF/VEGFR2/ZO-1 signaling pathway

Yunfang Yang  1 , Changjiang Li  1 , Sijin Yang  1 , Zhuo Zhang  2 , Xue Bai  1 , Hongmei Tang  1 , Jiang Huang  3
Affiliations

Cepharanthine maintains integrity of the blood-brain barrier (BBB) in stroke via the VEGF/VEGFR2/ZO-1 signaling pathway

Yunfang Yang et al. Aging (Albany NY). .

Abstract

Dysfunction of tight junctions such as zonula occludens protein-1 (ZO-1)-associated aggravation of blood-brain barrier (BBB) permeability plays an important role in the progression of stroke. Cepharanthine (CEP) is an extract from the plant Stephania cepharantha. However, the effects of CEP on stroke and BBB dysfunction have not been previously reported. In this study, we report that CEP improved dysfunction in neurological behavior in a middle cerebral artery occlusion (MCAO) mouse model. Importantly, CEP suppressed blood-brain barrier (BBB) hyperpermeability by increasing the expression of ZO-1. Notably, we found that CEP inhibited the expression of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor 2 (VEGFR2) in the cortex of MCAO mice. Additionally, the results of in vitro experiments demonstrate that treatment with CEP ameliorated cytotoxicity of human bEnd.3 brain microvascular endothelial cells against hypoxia/reperfusion (H/R). Also, CEP attenuated H/R-induced aggravation of endothelial permeability in bEND.3 cells by restoring the expression of ZO-1. Further study proved that the protective effects of CEP are mediated by inhibition of VEGF-A and VEGFR2. Based on the results, we conclude that CEP might possess a therapeutic prospect in stroke through protecting the integrity of the BBB mediated by the VEGF/VEGFR2/ZO-1 axis.

Keywords: Cepharanthine; blood-brain barrier (BBB); hypoxia/reperfusion (H/R); stroke; zonula occludens-1 (ZO-1).

PubMed Disclaimer

Conflict of interest statement

CONFLICTS OF INTEREST: All authors declared that there are no financial interests that are directly or indirectly related to the work submitted for publication.

Figures

Figure 1
Figure 1
Cepharanthine (CEP) improved neurological dysfunction in a middle cerebral artery occlusion (MCAO) mice model. (A) Molecular structure of Cepharanthine; (B) Neurological score graph of the three experimental groups (**, P<0.01 vs. vehicle group; ##, P<0.01 vs. MCAO group).
Figure 2
Figure 2
Cepharanthine (CEP) prevented the increase in blood-brain barrier (BBB) permeability in MCAO mice model. Blood-brain barrier permeability was measured by diffusion of sodium fluorescein assay (**, P<0.01 vs. vehicle group; ##, P<0.01 vs. MCAO group).
Figure 3
Figure 3
Cepharanthine (CEP) restored the expression of ZO-1 in the cortex of MCAO mice model. (A) mRNA of ZO-1; (B) Protein of ZO-1 as measured using immunostaining. Scale bars, 100 μm (**, P<0.01 vs. vehicle group; ##, P<0.01 vs. MCAO group).
Figure 4
Figure 4
Cepharanthine (CEP) reduced the expression of VEGF-A and VEGFR2 in the cortex of MCAO mice model. (A) mRNA levels of VEGF-A and VEGFR2; (B) Protein levels of VEGF-A and VEGFR2 as measured using immunostaining. Scale bars, 100 μm (**, P<0.01 vs. vehicle group; ##, P<0.01 vs. MCAO group).
Figure 5
Figure 5
Cepharanthine (CEP) ameliorated cytotoxicity of human bEnd.3 brain microvascular endothelial cells against hypoxia/reperfusion (H/R). Cells were exposed to hypoxia/reperfusion condition (6 h/24 h) with or without CEP (1.5, 3 μM). (A) Cell viability measured by MTT assay; (B) LDH release (**, P<0.01 vs. vehicle group; #, ##, P<0.05, 0.01 vs. H/R group).
Figure 6
Figure 6
Cepharanthine (CEP) attenuated hypoxia/reperfusion-induced aggravation of endothelial permeability in brain bEND.3 endothelial cells. (A) Endothelial permeability was measured using FITC-dextran; (B) The trans-endothelial electrical resistance (TEER) (**, P<0.01 vs. vehicle group; #, ##, P<0.05, 0.01 vs. H/R group).
Figure 7
Figure 7
Cepharanthine (CEP) restored the expression of ZO-1 in brain bEND.3 endothelial cells. (A) mRNA of ZO-1; (B) Protein of ZO-1 as measured by western blot analysis (**, P<0.01 vs. vehicle group; #, ##, P<0.05, 0.01 vs. H/R group).
Figure 8
Figure 8
Cepharanthine (CEP) reduced the expression of VEGF-A and VEGFR2 in brain bEND.3 endothelial cells. (A) mRNA of VEGF-A and VEGFR2; (B) Protein of VEGF-A and VEGFR2 (**, P<0.01 vs. vehicle group; #, ##, P<0.05, 0.01 vs. H/R group).
Figure 9
Figure 9
VEGF-A abolished the protective effects of CEP on the expression of ZO-1 and endothelial permeability. Cells were exposed to OGD/R in the presence or absence of CEP (3 μM) or VEGF-A (10 ng/ml). (A) mRNA of ZO-1; (B) Endothelial permeability was measured using FITC-dextran; (C) The trans-endothelial electrical resistance (TEER) (**, P<0.01 vs. vehicle group; ##, P<0.01 vs. H/R group group; ^, P<0.05 vs. H/R+CEP group).
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. George PM, Steinberg GK. Novel Stroke Therapeutics: Unraveling Stroke Pathophysiology and Its Impact on Clinical Treatments. Neuron. 2015; 87:297–309. 10.1016/j.neuron.2015.05.041 - DOI - PMC - PubMed
    1. Abbott NJ, Patabendige AA, Dolman DE, Yusof SR, Begley DJ. Structure and function of the blood-brain barrier. Neurobiol Dis. 2010; 37:13–25. 10.1016/j.nbd.2009.07.030 - DOI - PubMed
    1. Liebner S, Czupalla CJ, Wolburg H. Current concepts of blood-brain barrier development. Int J Dev Biol. 2011; 55:467–76. 10.1387/ijdb.103224sl - DOI - PubMed
    1. Ballabh P, Braun A, Nedergaard M. The blood-brain barrier: an overview: structure, regulation, and clinical implications. Neurobiol Dis. 2004; 16:1–13. 10.1016/j.nbd.2003.12.016 - DOI - PubMed
    1. Fujimura M, Gasche Y, Morita-Fujimura Y, Massengale J, Kawase M, Chan PH. Early appearance of activated matrix metalloproteinase-9 and blood-brain barrier disruption in mice after focal cerebral ischemia and reperfusion. Brain Res. 1999; 842:92–100. 10.1016/s0006-8993(99)01843-0 - DOI - PubMed

Publication types

MeSH terms

-