Cultured cells of the blood-brain barrier from apolipoprotein B-100 transgenic mice: effects of oxidized low-density lipoprotein treatment

• Published in: Fluids and barriers of the CNS 2015-07, Vol.12 (1), p.17, Article 17
• Main Authors: Lénárt, Nikolett, Walter, Fruzsina R, Bocsik, Alexandra, Sántha, Petra, Tóth, Melinda E, Harazin, András, Tóth, Andrea E, Vizler, Csaba, Török, Zsolt, Pilbat, Ana-Maria, Vígh, László, Puskás, László G, Sántha, Miklós, Deli, Mária A
• Format: Article
• Language: English
• Subjects: Animals; Apolipoprotein B-100 - genetics; Apolipoprotein B-100 - metabolism; Apolipoproteins; Atherosclerosis; Atherosclerosis - metabolism; Blood-Brain Barrier - cytology; Blood-Brain Barrier - drug effects; Blood-Brain Barrier - physiology; Brain; Care and treatment; Cell Survival - drug effects; Cells, Cultured; Disease Models, Animal; Endothelial Cells - drug effects; Endothelial Cells - metabolism; Endothelium; Genetic engineering; Immunohistochemistry; Lipoproteins, LDL - toxicity; Low density lipoproteins; Membrane Fluidity - drug effects; Mice; Mice, Inbred C57BL; Mice, Transgenic; Nervous system diseases; Neuroglia - drug effects; Neuroglia - metabolism; Nitric oxide; Nitric Oxide - metabolism; Pericytes - drug effects; Pericytes - metabolism; Permeability; Physiological aspects; Reactive Oxygen Species - metabolism
• ISSN: 2045-8118; 2045-8118
• DOI: 10.1186/s12987-015-0013-y

The apolipoprotein B-100 (ApoB-100) transgenic mouse line is a model of human atherosclerosis. Latest findings suggest the importance of ApoB-100 in the development of neurodegenerative diseases and microvascular/perivascular localization of ApoB-100 protein was demonstrated in the cerebral cortex of ApoB-100 transgenic mice. The aim of the study was to characterize cultured brain endothelial cells, pericytes and glial cells from wild-type and ApoB-100 transgenic mice and to study the effect of oxidized low-density lipoprotein (oxLDL) on these cells. Morphology of cells isolated from brains of wild type and ApoB-100 transgenic mice was characterized by immunohistochemistry and the intensity of immunolabeling was quantified by image analysis. Toxicity of oxLDL treatment was monitored by real-time impedance measurement and lactate dehydrogenase release. Reactive oxygen species and nitric oxide production, barrier permeability in triple co-culture blood-brain barrier model and membrane fluidity were also determined after low-density lipoprotein (LDL) or oxLDL treatment. The presence of ApoB-100 was confirmed in brain endothelial cells, while no morphological change was observed between wild type and transgenic cells. Oxidized but not native LDL exerted dose-dependent toxicity in all three cell types, induced barrier dysfunction and increased reactive oxygen species (ROS) production in both genotypes. A partial protection from oxLDL toxicity was seen in brain endothelial and glial cells from ApoB-100 transgenic mice. Increased membrane rigidity was measured in brain endothelial cells from ApoB-100 transgenic mice and in LDL or oxLDL treated wild type cells. The morphological and functional properties of cultured brain endothelial cells, pericytes and glial cells from ApoB-100 transgenic mice were characterized and compared to wild type cells for the first time. The membrane fluidity changes in ApoB-100 transgenic cells related to brain microvasculature indicate alterations in lipid composition which may be linked to the partial protection against oxLDL toxicity.