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Ruscogenin Attenuates Cerebral Ischemia-Induced Blood-Brain Barrier Dysfunction by Suppressing TXNIP/NLRP3 Inflammasome Activation and the MAPK Pathway

View Article: PubMed Central - PubMed

ABSTRACT

Ruscogenin, an important steroid sapogenin derived from Ophiopogon japonicus, has been shown to inhibit cerebral ischemic injury. However, its potential molecular action on blood-brain barrier (BBB) dysfunction after stroke remains unclear. This study aimed to investigate the effects of ruscogenin on BBB dysfunction and the underlying mechanisms in middle cerebral artery occlusion/reperfusion (MCAO/R)-injured mice and oxygen–glucose deprivation/reoxygenation (OGD/R)-injured mouse brain microvascular endothelial cells (bEnd.3). The results demonstrated that administration of ruscogenin (10 mg/kg) decreased the brain infarction and edema, improved neurological deficits, increased cerebral brain flow (CBF), ameliorated histopathological damage, reduced evans blue (EB) leakage and upregulated the expression of tight junctions (TJs) in MCAO/R-injured mice. Meanwhile, ruscogenin (0.1–10 µM) treatment increased cell viability and trans-endothelial electrical resistance (TEER) value, decreased sodium fluorescein leakage, and modulated the TJs expression in OGD/R-induced bEnd.3 cells. Moreover, ruscogenin also inhibited the expression of interleukin-1β (IL-1β) and caspase-1, and markedly suppressed the expression of Nucleotide-binding domain (NOD)-like receptor family, pyrin domain containing 3 (NLRP3) and thiredoxin-interactive protein (TXNIP) in vivo and in vitro. Furthermore, ruscogenin decreased reactive oxygen species (ROS) generation and inhibited the mitogen-activated protein kinase (MAPK) pathway in OGD/R-induced bEnd.3 cells. Our findings provide some new insights into its potential application for the prevention and treatment of ischemic stroke.

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Effects of ruscogenin on BBB permeability and the expression of tight junction proteins in mice following MCAO/R. (A,B) Representative gross appearance of EB-stained brain section in mice and quantitative analysis of EB extravasation by spectrophotometry (n = 6); (C,D) representative Western blot bands and quantitative analysis of the expression levels of ZO-1 and occludin in different groups. The band intensities were measured using scanning densitometry. The data were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression (n = 3). NS means “not significant”. The data are expressed as means ± SD. ##p < 0.01 vs. Sham, * p < 0.05, ** p < 0.01 vs. Model.
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ijms-17-01418-f003: Effects of ruscogenin on BBB permeability and the expression of tight junction proteins in mice following MCAO/R. (A,B) Representative gross appearance of EB-stained brain section in mice and quantitative analysis of EB extravasation by spectrophotometry (n = 6); (C,D) representative Western blot bands and quantitative analysis of the expression levels of ZO-1 and occludin in different groups. The band intensities were measured using scanning densitometry. The data were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression (n = 3). NS means “not significant”. The data are expressed as means ± SD. ##p < 0.01 vs. Sham, * p < 0.05, ** p < 0.01 vs. Model.

Mentions: BBB permeability was evaluated by the EB staining in the brains of mice, and the result of the quantitative spectrometry detected EB leakage in the model group and showed a significant EB extravasation compared with sham group and sham + RUS group at 24 h after reperfusion. Ruscogenin (10 mg/kg) treatment significantly reduced the leaked EB content (Figure 3A,B). We also investigated the effects of ruscogenin on the expression of ZO-1 and occludin at 24 h after reperfusion. Western blot analysis indicated that ruscogenin treatment could upregulate the decreased expressions of ZO-1 and occludin after ischemic stroke in mice (Figure 3C,D). No obvious change was observed between Sham and Sham + RUS in the above experiments.


Ruscogenin Attenuates Cerebral Ischemia-Induced Blood-Brain Barrier Dysfunction by Suppressing TXNIP/NLRP3 Inflammasome Activation and the MAPK Pathway
Effects of ruscogenin on BBB permeability and the expression of tight junction proteins in mice following MCAO/R. (A,B) Representative gross appearance of EB-stained brain section in mice and quantitative analysis of EB extravasation by spectrophotometry (n = 6); (C,D) representative Western blot bands and quantitative analysis of the expression levels of ZO-1 and occludin in different groups. The band intensities were measured using scanning densitometry. The data were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression (n = 3). NS means “not significant”. The data are expressed as means ± SD. ##p < 0.01 vs. Sham, * p < 0.05, ** p < 0.01 vs. Model.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5037697&req=5

ijms-17-01418-f003: Effects of ruscogenin on BBB permeability and the expression of tight junction proteins in mice following MCAO/R. (A,B) Representative gross appearance of EB-stained brain section in mice and quantitative analysis of EB extravasation by spectrophotometry (n = 6); (C,D) representative Western blot bands and quantitative analysis of the expression levels of ZO-1 and occludin in different groups. The band intensities were measured using scanning densitometry. The data were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression (n = 3). NS means “not significant”. The data are expressed as means ± SD. ##p < 0.01 vs. Sham, * p < 0.05, ** p < 0.01 vs. Model.
Mentions: BBB permeability was evaluated by the EB staining in the brains of mice, and the result of the quantitative spectrometry detected EB leakage in the model group and showed a significant EB extravasation compared with sham group and sham + RUS group at 24 h after reperfusion. Ruscogenin (10 mg/kg) treatment significantly reduced the leaked EB content (Figure 3A,B). We also investigated the effects of ruscogenin on the expression of ZO-1 and occludin at 24 h after reperfusion. Western blot analysis indicated that ruscogenin treatment could upregulate the decreased expressions of ZO-1 and occludin after ischemic stroke in mice (Figure 3C,D). No obvious change was observed between Sham and Sham + RUS in the above experiments.

View Article: PubMed Central - PubMed

ABSTRACT

Ruscogenin, an important steroid sapogenin derived from Ophiopogon japonicus, has been shown to inhibit cerebral ischemic injury. However, its potential molecular action on blood-brain barrier (BBB) dysfunction after stroke remains unclear. This study aimed to investigate the effects of ruscogenin on BBB dysfunction and the underlying mechanisms in middle cerebral artery occlusion/reperfusion (MCAO/R)-injured mice and oxygen&ndash;glucose deprivation/reoxygenation (OGD/R)-injured mouse brain microvascular endothelial cells (bEnd.3). The results demonstrated that administration of ruscogenin (10 mg/kg) decreased the brain infarction and edema, improved neurological deficits, increased cerebral brain flow (CBF), ameliorated histopathological damage, reduced evans blue (EB) leakage and upregulated the expression of tight junctions (TJs) in MCAO/R-injured mice. Meanwhile, ruscogenin (0.1&ndash;10 &micro;M) treatment increased cell viability and trans-endothelial electrical resistance (TEER) value, decreased sodium fluorescein leakage, and modulated the TJs expression in OGD/R-induced bEnd.3 cells. Moreover, ruscogenin also inhibited the expression of interleukin-1&beta; (IL-1&beta;) and caspase-1, and markedly suppressed the expression of Nucleotide-binding domain (NOD)-like receptor family, pyrin domain containing 3 (NLRP3) and thiredoxin-interactive protein (TXNIP) in vivo and in vitro. Furthermore, ruscogenin decreased reactive oxygen species (ROS) generation and inhibited the mitogen-activated protein kinase (MAPK) pathway in OGD/R-induced bEnd.3 cells. Our findings provide some new insights into its potential application for the prevention and treatment of ischemic stroke.

No MeSH data available.


Related in: MedlinePlus