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Protein expression of HIF-1 alpha, VEGF and cyclooxygenases in cerebral blood vessels of Sprague- Dawley rats on a short-term high salt diet

Question: Our previous study showed increased oxidative stress and impaired vascular reactivity in cerebral blood vessels of rats on high salt diet (HSD). Increased oxidative stress can affect HIF-1α expression and its downstream proteins, such as COX- 2. Purpose of this experiment was to determine effect of HS diet on protein expression of HIF-1α, its target protein VEGF and COX-1 and COX-2 in cerebral blood vessels. Methods: 11 week old Sprague-Dawley rats (N=8 per group) were divided in three groups: CTRL (control group of rat) ; HSD (rats on HS diet for 7 days, 4% NaCl) and HSD+TEMPOL (rats on HS diet and 1mM TEMPOL in drinking water for 7 days). Prior to decapitation and surface cerebral blood vessels collection, rats were anesthetized with ketamin-chlorid (75 mg/kg) and midazolam (0.5 mg/kg). Protein levels of HIF-1α, VEGF and COX-1 and COX-2 were assesed. One-way ANOVA or student t-test was used when appropriate (SigmaPlot v11.2, Systat Software, Chicago, USA), 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 the local and national Ethical Committee. Results: Results show significantly increased expression of HIF-1alpha transcriptional factor and VEGF in HSD groups compared to CTRL and HSD+TEMPOL group. There was no differences in COX-1 expression. Expression of COX-2 was increased in HSD group compared to HSD+TEMPOL group. Conclusion: Increased protein levels of HIF-1alpha and COX-2 in brain blood vessels may be related to increased oxidative stress caused by high salt diet and reveresed by TEMPOL in vivo. Acknowledgements: Supported by Croatian Science Foundation under the project #IP-2014-09-6380.

hih-salt diet, Sprague-Dawley rats, HIF-1 alpha, VEGF, cyclooxygenases, cerebral blood vessels

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