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

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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 the TXNIP/NLRP3 pathway in bEnd.3 cells subjected to OGD/R. (A–F) Representative Western blot bands and quantitative analysis of the ratio of IL-1β and pro-IL-1β, caspase-1 and pro-caspase-1, NLRP3 and TXNIP. The band intensities were measured using scanning densitometry. The data were normalized to GAPDH expression (n = 3). The data are expressed as means ± SD. ##p < 0.01 vs. Control, * p < 0.05, ** p < 0.01 vs. Model.
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ijms-17-01418-f007: Effects of ruscogenin on the TXNIP/NLRP3 pathway in bEnd.3 cells subjected to OGD/R. (A–F) Representative Western blot bands and quantitative analysis of the ratio of IL-1β and pro-IL-1β, caspase-1 and pro-caspase-1, NLRP3 and TXNIP. The band intensities were measured using scanning densitometry. The data were normalized to GAPDH expression (n = 3). The data are expressed as means ± SD. ##p < 0.01 vs. Control, * p < 0.05, ** p < 0.01 vs. Model.

Mentions: We also investigated the expression of IL-1β, caspase-1 and the TXNIP/NLRP3 pathway in bEnd.3 cells subjected to 6 h of OGD and 18 h reoxygenation. The Western blot results demonstrated that pretreatment with ruscogenin could downregulate the increased expression of IL-1β and caspase-1 proteins, and inhibited the expressions of NLRP3 and TXNIP (Figure 7A–F).


Ruscogenin Attenuates Cerebral Ischemia-Induced Blood-Brain Barrier Dysfunction by Suppressing TXNIP/NLRP3 Inflammasome Activation and the MAPK Pathway
Effects of ruscogenin on the TXNIP/NLRP3 pathway in bEnd.3 cells subjected to OGD/R. (A–F) Representative Western blot bands and quantitative analysis of the ratio of IL-1β and pro-IL-1β, caspase-1 and pro-caspase-1, NLRP3 and TXNIP. The band intensities were measured using scanning densitometry. The data were normalized to GAPDH expression (n = 3). The data are expressed as means ± SD. ##p < 0.01 vs. Control, * p < 0.05, ** p < 0.01 vs. Model.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5037697&req=5

ijms-17-01418-f007: Effects of ruscogenin on the TXNIP/NLRP3 pathway in bEnd.3 cells subjected to OGD/R. (A–F) Representative Western blot bands and quantitative analysis of the ratio of IL-1β and pro-IL-1β, caspase-1 and pro-caspase-1, NLRP3 and TXNIP. The band intensities were measured using scanning densitometry. The data were normalized to GAPDH expression (n = 3). The data are expressed as means ± SD. ##p < 0.01 vs. Control, * p < 0.05, ** p < 0.01 vs. Model.
Mentions: We also investigated the expression of IL-1β, caspase-1 and the TXNIP/NLRP3 pathway in bEnd.3 cells subjected to 6 h of OGD and 18 h reoxygenation. The Western blot results demonstrated that pretreatment with ruscogenin could downregulate the increased expression of IL-1β and caspase-1 proteins, and inhibited the expressions of NLRP3 and TXNIP (Figure 7A–F).

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