Transgenic rat with overproduction of ubiquitous angiotensin-(1-7) presents neuroprotection in a model of ischemia and reperfusion

Lucas Miranda Kangussu; Ana Flávia Almeida-Santos; Lorena Figueiredo Fernandes; Natalia Alenina; Michael Bader; Robson A S Santos; André Ricardo Massensini; Maria José Campagnole-Santos

doi:10.1016/j.brainresbull.2022.11.017

Transgenic rat with overproduction of ubiquitous angiotensin-(1-7) presents neuroprotection in a model of ischemia and reperfusion

Brain Res Bull. 2023 Jan:192:184-191. doi: 10.1016/j.brainresbull.2022.11.017. Epub 2022 Nov 23.

Authors

Lucas Miranda Kangussu¹, Ana Flávia Almeida-Santos², Lorena Figueiredo Fernandes², Natalia Alenina³, Michael Bader⁴, Robson A S Santos², André Ricardo Massensini², Maria José Campagnole-Santos⁵

Affiliations

¹ Department of Morphology - Biological Sciences Institute, Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, Brazil. Electronic address: lucaskangussu@ufmg.br.
² Department of Physiology and Biophysics - Biological Sciences Institute, Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, Brazil.
³ Max-Delbrück Center for Molecular Medicine (MDC), Berlin, Germany; German Center for Cardiovascular Research (DZHK), Partner Site Berlin, Berlin, Germany.
⁴ Max-Delbrück Center for Molecular Medicine (MDC), Berlin, Germany; German Center for Cardiovascular Research (DZHK), Partner Site Berlin, Berlin, Germany; Charité University Medicine Berlin, Berlin, Germany; Institute for Biology, University of Lübeck, Lübeck, Germany.
⁵ Department of Physiology and Biophysics - Biological Sciences Institute, Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, Brazil. Electronic address: mjcampagnole@ufmg.br.

PMID: 36435363
DOI: 10.1016/j.brainresbull.2022.11.017

Abstract

Recent studies showed that angiotensin-(1-7) has cerebroprotective actions in stroke. In the present study, we aim to test whether tissue overexpression of Angiotensin-(1-7), mainly in the brain provides neuroprotection in a model of ischemia/reperfusion by bilateral common carotid arteries occlusion/reperfusion (BCCAo/R). Evaluation of neurological deficit scores and bilateral asymmetry test (BAT) were performed seven days after transient BCCAo/R in transgenic rats (TG-7371) overexpressing Angiotensin-(1-7) and Sprague-Dawley (SD) rats. To assess blood-brain barrier (BBB) permeability Evans blue dye (EB) was intravenously injected. Cytokine levels were quantified in the whole brain through Elisa assay and oxidative stress was measured 7 days after ischemia. The expression of AT₁ and Mas receptors and inducible nitric oxide synthase (iNOS) was evaluated by RT-PCR. Neurological deficits were observed in both SD-BCCAo/R and TG-BCCAo/R, contrasting to sham-operated groups. However, TG-BCCAo/R showed a significant lower neurological score and latency in BAT when compared with SD-BCCAo/R. BBB integrity in TG-BCCAo/R was improved, since these animals showed lower extravasation of EB than SD-BCCAo/R. Interestingly, TG-BCCAo/R presented lower levels of pro-inflammatory cytokines when compared to SD-BCCAo/R. Levels of IL-10 were higher in SD-BCCAo/R than in SD control and even higher in TG-BCCAo/R. TG-BCCAo/R animals presented decreased levels of TBARS and increase in SOD activity and GSH levels when compared to SD sham rats. RT-PCR results showed higher levels of AT₁ receptor and iNOS in SD-BCCAo/R compared to TG-BCCAo/R, but no difference was observed for Mas receptor. The present study shows that lifetime increase in cerebral expression of an Ang-(1-7)-producing fusion protein induces neuroprotection in experimental global cerebral ischemia and reperfusion, reassuring that, pharmacological strategies leading to increase in Ang-(1-7) can be an additional tool for stroke therapy.

Keywords: Angiotensin-(1-7); Ischemia; Neuroprotection; Stroke; TG-7371; Transgenic rats.

MeSH terms

Animals
Brain Ischemia* / metabolism
Ischemia
Rats
Rats, Sprague-Dawley
Rats, Transgenic
Reperfusion Injury* / metabolism
Stroke*

Substances

angiotensin I (1-7)