engleski

Establishing the model of tMCAO induced stroke in TFF3-/- knockout mice

Arachidonic acid (AA) metabolismand products affect vascular reactivity mechanism. TFF3-/- mice have protective phenotype of favorable ratio of ω-6/ω-3 fatty acids and modified metabolism of AA. However, the role of TFF3 peptide in hypertension, stroke and vascular function is still underinvestigated. High salt (HS) intake is one of the major risk factors for the development and progression of arterial hypertension directly proportional to the risk of stroke. This study aimed to establish the method of transient middle cerebral artery (t- MCAO) occlusion-reperfusion stroke model in WT and TFF3-/-mice in our laboratory and to asses the effect of HS diet on stroke volume. WT and TFF3-/-mice aged 12-14 weeks were divided into 4 groups: (WT_LS, TFF3-/-_LS-fed standard chow) ; and groups fed HS (4% NaCl) diet for 7 days prior the surgery, WT_HSD and TFF3-/-_HSD. Animals were anaesthetized with 2% isoflurane and heated to avoid hypothermia. After midline neck incision, silicon coated 6-0 monofilament was inserted via external carotid artery into internal carotid artery, and then into the circle of Willis, thus occluding MCA for 60 min.n 24h after surgery mice were anaesthetized with ketamine and midazolam and decapitated. The brains were cut into 2-mm-thick coronal sections, stained with 2% 2, 3, 5- triphenyltetrazolium chloride, fixed in 10% buffered formalin solution and scanned. Infarct volumes were measured using ImageJ imaging software and expressed as a percentage of the ischaemic hemisphere. All experimental procedures conformed to the European Guidelines for the Care and Use of Laboratory Animals (directive 86/609) and were approved by institutional Ethical Committee. Result of this pilot study (N=3 /group) show successfully established model of tMCAO in mice, but no significant differencies in cortical and total infarct size, due to small sample. Partially supported by Croatian Science Foundation under the project #IP-2014-09-6380: “Impaired Vasorelaxation and Endothelial Leukocyte Interaction (ELI) in Development of Atherosclerotic Lesions -V-ELI Athero“ and Faculty of Medicine Osijek VIF2018-MEFOS-09- 1509 grant (PI Drenjancevic I).

transient middle cerebral artery (t-MCAO) ; TFF3-/- knockout mice

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