Abstract
The Neurovascular Unit (NVU) is formed by vascular and neural cells controlling the cerebral hyperaemia. All the components are anatomically and functionally linked to each other, resulting in a highly efficient regulation of the cerebral blood flow, which, when interrupted, can lead to stroke. An ischemic stroke (IS) is the most common type of stroke with high rates of morbidity, mortality and disability. Therefore, it is of extreme importance to protect the functional and structural integrity of the NVU in patients with IS, understanding the mechanisms involved and how it affects each component of the NVU. Thus, the aim of this work is to analyse how the vascular smooth muscle cells from the rat middle cerebral artery function/react after an ischemic event. To mimic this event, primary cortical cultures were challenged to oxygen and glucose deprivation (OGD) for 4 h and 6 h, and the smooth muscle cells (SMCs) contractility was analysed after exposure to different media previously conditioned by the cortical cultures upon reperfusion. The results show a dual effect on the SMCs response to the vasorelaxant agent, only for cells exposed to the reperfusion media conditioned by neuron-glia cultures challenged by OGD, leading to increased relaxation of the SMCs for OGD 4 h, whereas for OGD 6 h the effect is reversed leading to contraction of the SMCs. These differences demonstrate that the astrocytes mediate the vasoactive response of vascular smooth muscle by releasing factors into the reperfusion medium, and the hypoxia time is fundamental for a beneficial/harmful response by the vascular smooth muscle.
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Acknowledgements
This work was also supported by FEDER funds through the POCI -COMPETE 2020—Operational Programme Competitiveness and Internationalisation in Axis I -Strengthening research, technological development, and innovation (Project POCI-01-0145-FEDER007491) and National Funds by FCT—Foundation for Science and Technology (Project UID/Multi/00709/2019), and by ‘‘Programa Operacional do Centro, Centro 2020” through the funding of the ICON project (Interdisciplinary Challenges On Neurodegeneration; CENTRO-01-0145-FEDER-000013)”.
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Elisa Cairrao conceived the study and designed the protocol. Melissa Mariana performed the experiments. Claudio Roque performed the primary cortical cultures. Elisa Cairrao supervised the study. Melissa Mariana and Elisa Cairrao analysed data. Melissa Mariana wrote the manuscript. Graça Baltazar and Elisa Cairrao revised the manuscript for intellectual content. Melissa Mariana, Claudio Roque, Graça Baltazar and Elisa Cairrao read, reviewed and edited the manuscript. All authors have approved the final version of the paper.
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The animals were used in accordance with the national ethical requirements for animal research and with the European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Scientific Purposes (Directive 2010/63/EU). All animal experiments were approved by the Animal Research Committee of University of Beira Interior (CICS-UBI, Covilhã, Portugal).
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Mariana, M., Roque, C., Baltazar, G. et al. In Vitro Model for Ischemic Stroke: Functional Analysis of Vascular Smooth Muscle Cells. Cell Mol Neurobiol 42, 2289–2304 (2022). https://doi.org/10.1007/s10571-021-01103-5
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DOI: https://doi.org/10.1007/s10571-021-01103-5