A Novel Role of Prolidase in Cocaine-Mediated Breach in the Barrier of Brain Microvascular Endothelial Cells

Aim: The aim of the current work is to assess the role of proBDNF/BDNF in the interaction between brain microvascular endothelial cells and MDA-MB-231 breast cancer cell line that has been consistently reported to cause brain metastasis. Background: Breast cancer brain metastasis (BM) is a significant health problem with limited therapeutic options. The development of BM is a multistep process that requires constant interaction with brain vasculature and development of tumor blood supply. The benefits of antiangiogenic modalities based on antagonizing vascular endothelial growth factor in breast cancer metastasis did not prove to be effective. Brain derived neurotrophic factor (BDNF) is a neurotrophin with reported angiogenic effect. There is a lack of data regarding the involvement of BDNF in metastatic breast cancer interaction with brain microvascular endothelial cells (HBEC-5i). Objectives: To determine the role of the pro form of BDNF in the interaction between MDA-MB-231 cells and brain endothelial cells. Methods: using an adaptive transfer design, the cross talk between HBEC-5i and MDA-MB-231 was investigated. HBEC-5i were treated with MDA-MB-231-conditioned media and the involvement of BDNF/proBDNF in the interaction was assessed using both release and inhibitor-based assays in migration and in vitro tube formation assay. Results: MDA-MB-231 and HBEC-5i released total BDNF (250 vs. 80 pg/ml, respectively). MDA-MB-231 conditioned media inhibited the migration of HBEC-5i by more than 80% (p<0.05) and tube formation by 75% (p<0.05). Neutralizing mature BDNF did not alter the MDA-MB-231 induced antiangiogenic effect which was completely blunted by antagonizing proBDNF. MDA-MB-231 released proBDNF (131.5 pg/ml) and more that 60% of total BDNF released was in the pro-form. Conclusion: proBDNF is a novel mediator of the breast cancer induced antiangiogenic effect in brain endothelial cells.

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The Role of Sphingomyelin Metabolism in the Protection of Rat Brain Microvascular Endothelial Cells by Mild Hypothermia

Neurocritical Care ◽

10.1007/s12028-021-01338-6 ◽

2021 ◽

Author(s):

Hai-hong Zhang ◽

Lin Wang ◽

Wei Zhang ◽

Zhi Wan

Keyword(s):

Endothelial Cells ◽

Rat Brain ◽

Mild Hypothermia ◽

Microvascular Endothelial Cells ◽

Brain Microvascular Endothelial Cells ◽

Microvascular Endothelial

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Age-related Beta-synuclein Alters the p53/Mdm2 Pathway and Induces the Apoptosis of Brain Microvascular Endothelial Cells In Vitro

Cell Transplantation ◽

10.1177/0963689718755706 ◽

2018 ◽

Vol 27 (5) ◽

pp. 796-813 ◽

Cited By ~ 4

Author(s):

Katrin Brockhaus ◽

Michael R. R. Böhm ◽

Harutyun Melkonyan ◽

Solon Thanos

Keyword(s):

Endothelial Cells ◽

Neurodegenerative Diseases ◽

Microvascular Endothelial Cells ◽

Mrna Levels ◽

Brain Microvascular Endothelial Cells ◽

Age Related ◽

Microvascular Endothelial ◽

Dose Dependent

Increased β-synuclein (Sncb) expression has been described in the aging visual system. Sncb functions as the physiological antagonist of α-synuclein (Snca), which is involved in the development of neurodegenerative diseases, such as Parkinson’s and Alzheimer’s diseases. However, the exact function of Sncb remains unknown. The aim of this study was to elucidate the age-dependent role of Sncb in brain microvascular endothelial cells (BMECs). BMECs were isolated from the cortices of 5- to 9-d-old Sprague-Dawley rats and were cultured with different concentrations of recombinant Sncb (rSncb) up to 72 h resembling to some degree age-related as well as pathophysiological conditions. Viability, apoptosis, expression levels of Snca, and the members of phospholipase D2 (Pld2)/ p53/ Mouse double minute 2 homolog (Mdm2)/p19(Arf) pathway, response in RAC-alpha serine/threonine-protein kinase (Akt), and stress-mediating factors such as heme oxygenase (decycling) 1 (Hmox) and Nicotinamide adenine dinucleotide phosphate oxygenase 4 (Nox4) were examined. rSncb-induced effects were confirmed through Sncb small interfering RNA (siRNA) knockdown in BMECs. We demonstrated that the viability decreases, while the rate of apoptosis underly dose-dependent alterations. For example, apoptosis increases in BMECs following the treatment with higher dosed rSncb. Furthermore, we observed a decrease in Snca immunostaining and messenger RNA (mRNA) levels following the exposure to higher rScnb concentrations. Akt was shown to be downregulated and pAkt upregulated by this treatment, which was accompanied by a dose-independent increase in p19(Arf) levels and enhanced intracellular Mdm2 translocation in contrast to a dose-dependent p53 activation. Moreover, Pld2 activity was shown to be induced in rSncb-treated BMECs. The expression of Hmox and Nox4 after Sncb treatment was altered on BEMCs. The obtained results demonstrate dose-dependent effects of Sncb on BMECs in vitro. For example, the p53-mediated and Akt-independent apoptosis together with the stress-mediated response of BMECs related to exposure of higher SNCB concentrations may reflect the increase in Sncb with duration of culture as well as its impact on cell decay. Further studies, expanding on the role of Sncb, may help understand its role in the neurodegenerative diseases.

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