engleski

Effect of high salt diet on vascular reactive oxygen species production in Tff3−/−/C57BL/6N knockout mice

Introduction: Tff3 gene knockout mice (Tff3−/ −/C57BL/6N) have changes in lipid metabolism which may affect vascular function (1, 2). This study aimed to asses the effect of high salt (HS) diet on endothelial reactive oxygen species (ROS) production in carotid arteries of Tff3-/- knockout mice and their wild type controls (WT, C57BL/6N). Methods: Male, ten-weeks-old transgenic Tff3−/ −/C57BL/6N and WT/C57BL/6N (parental strain) healthy mice were divided in LS (0.4% NaCl) and HS (4% NaCl in rodent chow fed for 1 week) groups. After anaesthesia (ketamine-chloride and midazolam), mice were decapitated and carotid arteries were isolated and cannulated on pressure myograph with (∆80 mmHg) or without flow (∆0 mmHg), in the absence/presence of the superoxide dismutase mimetic TEMPOL (100 μmol l−1). ROS production was determined by dihydroethidine (DHE, 20 μM) fluorescence assay. Statistical analyses were performed with One-way ANOVA test ; p<0.05 was considered significant. All experimental procedures conformed to the European Guidelines for the Care and Use of Laboratory Animals (directive 86/609) and were approved by local and national Ethical Committee (No.2158/61- 02-139/2-06 ; No.2158/61-07-14-119). Results: Basal (no-flow) ROS production was significantly increased in WT_ HS compared to the WT_ LS group and in Tff3−/−_HS compared to Tff3−/ −_LS group. Tempol significantly decreased ROS production in WT_ HS and Tff3−/−_HS groups. Under flow conditions, ROS production was significantly increased in WT_ HS compared to WT_LS group and in Tff3−/−_HS compared to Tff3−/ −_LS group. Tempol significantly decreased ROS production in all groups. Conclusion: HS diet increases generation of ROS and increases vascular oxidative stress in both strains. Acknowledgments: The author would like to thank all the members from the group Department of Physiology and Immunology, Faculty of Medicine Osijek. Financial support: This study was supported by the Croatian Science Foundation under the project IP- 2014-09-6380 (V-ELI Athero), VIF-2018-MEFOS-09- 1509 grant and Faculty of Medicine Osijek Institutional grant #IP-1-MEFOS2019 and #IP-2- MEFOS2020 (PI Ines Drenjančević).

Tff3 gene, high salt diet, reactive oxygen species, fluorescence

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