Edaravone Protects against Methylglyoxal-Induced Barrier Damage in Human Brain Endothelial Cells

Tóth, Anna Zsófia; Walter, Fruzsina R.; Bocsik, Alexandra; Sántha, Petra; Veszelka, Szilvia; Nagy, L; Puskás, László G.; Couraud, Pierre Olivier; Takata, Fuyuko; Dohgu, Shinya; Kataoka, Yasufumi; Deli, Mária A.

doi:10.1371/journal.pone.0100152

Cited by 35 publications

(43 citation statements)

References 67 publications

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“…Rat brain endothelial cells, astrocytes and pericytes were treated with kainate (213 Da; 100 mM stock solution was prepared in sterile water by the addition of NaOH) at 10 µM and 100 µM concentrations in culture medium for 1 and 24 h, based on our preliminary experiment and in agreement with literature data [32]. Simvastatin, edaravone (for both 1 mM stock solution was prepared in dimethyl sulfoxide) and dexamethasone (cyclodextrin complex; 10 mM stock solution was prepared in sterile water) were applied at a concentration of 1 µM based on our preliminary study and literature data [21,23,25]. The control group received culture medium.…”

Section: Treatmentsmentioning

confidence: 88%

“…This method has been established to follow not only cell attachment and growth, but cell viability as well. We have verified by endpoint colorimetric tests and morphological methods that impedance changes can sensitively detect cell damage [23,29,33] and protection [23,29]. The RTCA-SP system (ACEA Biosciences, CA, USA) registers the impedance of cells automatically every 10 min.…”

Section: Real-time Cell Analysismentioning

confidence: 96%

“…Edaravone is an excellent free radical scavenger molecule, which is clinically used for treating acute stroke and amyotrophic lateral sclerosis [17]. Our group demonstrated the protective effect of edaravone on brain endothelial cells against methylglyoxal-induced barrier damage [23]. In a kainate-induced epilepsy model in rats edaravone significantly decreased neuronal cell death and hyperexcitability [24].…”

Section: Introductionmentioning

confidence: 92%

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Simvastatin, edaravone and dexamethasone protect against kainate-induced brain endothelial cell damage

Barna

Walter

Harazin

et al. 2020

Fluids Barriers CNS

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Background: Excitotoxicity is a central pathological pathway in many neurological diseases with blood-brain barrier (BBB) dysfunction. Kainate, an exogenous excitotoxin, induces epilepsy and BBB damage in animal models, but the direct effect of kainate on brain endothelial cells has not been studied in detail. Our aim was to examine the direct effects of kainate on cultured cells of the BBB and to test three anti-inflammatory and antioxidant drugs used in clinical practice, simvastatin, edaravone and dexamethasone, to protect against kainate-induced changes.Methods: Primary rat brain endothelial cell, pericyte and astroglia cultures were used to study cell viability by impedance measurement. BBB permeability was measured on a model made from the co-culture of the three cell types. The production of nitrogen monoxide and reactive oxygen species was followed by fluorescent probes. The mRNA expression of kainate receptors and nitric oxide synthases were studied by PCR.Results: Kainate damaged brain endothelial cells and made the immunostaining of junctional proteins claudin-5 and zonula occludens-1 discontinuous at the cell border indicating the opening of the barrier. The permeability of the BBB model for marker molecules fluorescein and albumin and the production of nitric oxide in brain endothelial cells were increased by kainate. Simvastatin, edaravone and dexamethasone protected against the reduced cell viability, increased permeability and the morphological changes in cellular junctions caused by kainate. Dexamethasone attenuated the elevated nitric oxide production and decreased the inducible nitric oxide synthase (NOS2/iNOS) mRNA expression increased by kainate treatment. Conclusion:Kainate directly damaged cultured brain endothelial cells. Simvastatin, edaravone and dexamethasone protected the BBB model against kainate-induced changes. Our results confirmed the potential clinical usefulness of these drugs to attenuate BBB damage. Publisher's NoteSpringer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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Section: Treatmentsmentioning

confidence: 88%

Section: Real-time Cell Analysismentioning

confidence: 96%

Section: Introductionmentioning

confidence: 92%

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Simvastatin, edaravone and dexamethasone protect against kainate-induced brain endothelial cell damage

Barna

Walter

Harazin

et al. 2020

Fluids Barriers CNS

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“…Culture-based BBB models greatly differ in complexity. Immortalized cell lines and mono-cultures of brain endothelial cells can be used as simplified BBB models [9][10][11], but the barrier properties of primary cell-based co-culture models are better [2,12,13]. We established a syngeneic rat BBB model based on the co-culture of primary brain endothelial cells with pericytes and glial cells mimicking the in vivo anatomical position of the cells.…”

mentioning

confidence: 99%

A versatile lab-on-a-chip tool for modeling biological barriers

Walter

Valkai

Kincses

et al. 2016

Sensors and Actuators B: Chemical

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136

155

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Keywords: Microfluidics Lung and intestinal epithelial cells Endothelial cells Blood-brain barrier PermeabilityTrans-epithelial/endothelial electric resistance a b s t r a c t Models of biological barriers are important to study physiological functions, transport mechanisms, drug delivery and pathologies. However, there are only a few integrated biochips which are able to monitor several of the crucial parameters of cell-culture-based barrier models. The aim of this study was to design and manufacture a simple but versatile device, which allows a complex investigation of barrier functions. The following functions and measurements are enabled simultaneously: co-culture of 2 or 3 types of cells; flow of culture medium; visualization of the entire cell layer by microscopy; real-time transcellular electrical resistance monitoring; permeability measurements. To this end, a poly(dimethylsiloxane)-based biochip with integrated transparent gold electrodes and with a possibility to connect to a peristaltic pump was built. Unlike previous systems, the structure of the device allowed a constant visual observation of cell growth over the whole membrane surface. Morphological characterization of the layers was also accomplished by immunohistochemical staining. The chip was applied to monitor and characterize models of the intestinal and lung epithelial barriers, and the blood-brain barrier. The models were established using human Caco-2 intestinal and A549 lung epithelial cell lines, hCMEC/D3 human brain endothelial cell line and primary rat brain endothelial cells co-cultured with primary astrocytes and brain pericytes. This triple primary co-culture blood-brain barrier model was assembled on a lab-on-a-chip device and investigated under fluid flow for the first time. Such a versatile tool is expected to facilitate the kinetic investigation of various biological barriers.

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“…Immortalized human cerebrovascular endothelial cell line (hCMEC/D3) was used in this study as an in vitro model of human BBB (Manda et al, 2011; Toth et al, 2014). Cell monolayers (p28-31) were cultured in buffered endothelial basal media containing 5% FBS, antibiotics and all supplements provided in EGM™ 2MV BulletKit (Lonza), as described previously (Sajja et al, 2015).…”

Section: Methodsmentioning

confidence: 99%

Hyperglycemia exacerbates antiretroviral drug combination induced blood-brain barrier endothelial toxicity

et al. 2016

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In this study, we sought to investigate how concomitant hyperglycemia influences the impact of combination antiretroviral therapy on blood-brain barrier (BBB) endothelial function. Immortalized human brain microvascular endothelial cell line (hCMEC/D3) was exposed to azidothymidine (AZT; a nucleoside reverse transcriptase inhibitor) and/or indinavir (IND; protease inhibitor) in normal glycemic (5.5mM) or hyperglycemic (HG; 25mM) media containing D-glucose for 24-72h. Cellular reactive oxygen species (ROS) and mitochondria-specific superoxide levels were assayed in addition to membrane potential to determine the extent of mitochondrial dysfunction. Nrf2 expression was analyzed by immunofluorescence. Our results indicated a significant increase in BBB endothelial toxicity (decreased ATP) by HG and AZT+IND with progression of time (24-72h). Concurrent HG and antiviral drug combination synergistically elevated BBB endothelial ROS induced by either condition alone. Further, HG and AZT+IND mutually interact to elicit a pronounced increase in mitochondrial superoxide levels post 24h (vs. either condition alone or controls). In addition, HG and AZT+IND complemented each other to induce potential loss of mitochondrial membrane potential. While HG or AZT+IND alone for 24h increased Nrf2 nuclear distribution, co-exposure conditions induced a potential loss of Nrf2 expression/nuclear translocation in BBB endothelium. In summary, our data strongly suggest that antiretroviral drug combination potentially interacts with concomitant HG and triggers exacerbated mitochondrial dysfunction and BBB endothelial toxicity, possibly through dysregulation of Nrf2 signaling. Thus, this study warrants the critical need for safety evaluation and monitoring of neurovascular complications of HAART regimens in HIV-infected diabetic patient cohort.

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Edaravone Protects against Methylglyoxal-Induced Barrier Damage in Human Brain Endothelial Cells

Cited by 35 publications

References 67 publications

Simvastatin, edaravone and dexamethasone protect against kainate-induced brain endothelial cell damage

Simvastatin, edaravone and dexamethasone protect against kainate-induced brain endothelial cell damage

A versatile lab-on-a-chip tool for modeling biological barriers

Hyperglycemia exacerbates antiretroviral drug combination induced blood-brain barrier endothelial toxicity

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