V-003 | In vitro analysis of cross-talk between angiotensin-II receptors (AT1R and AT2R), neurotrophic factors and pro-inflammatory agents in the context of nerve regeneration.

Cellular and Molecular Neurobiology
Author: Sergio Gonzalo Benitez | Email: sbenitez595@gmail.com

Cristian Acosta^1°, Alicia Seltzer^1°,Sergio Benitez^1°

^1° Laboratorio de Estudios Neurobiológicos (LABENE). Instituto de Histología y Embriología de Mendoza (IHEM-CONICET), Universidad Nacional de Cuyo, 5502, Mendoza, Argentina.

There is evidence that the Renin Angiotensin system (RAS) is involved in the process of nerve regeneration. Therefore, understanding the complex interactions between RAS components and the repair and inflammation systems is essential to validate the use of angiotensinergic drugs as an effective therapeutic agent in nerve injuries. Using primary cultures of rat dorsal root ganglion cells, different treatments (inflammatory soup (IS), angiotensin-II, Azilsartan or PD123319) and culture times (2 or 3 days in vitro, DIV), we sought to study the interaction between trophic and inflammatory factors and the variations in the expression of AT1R and AT2R. Cultures were analyzed by qPCR and immunofluorescence (IF). Our results indicate that 2DIV cultures grown in the presence of both GDNF and NGF increased the expression levels of AT1R, whereas AT2R was only upregulated in the presence of IS for one day with NGF. IS alone increased the protein levels of AT1R and AT2R in the population of neurons identified by the expression of b-tubulin III (IF). However, at 3DIV IS alone increased the global expression of both AT1R and AT2R receptors (qPCR). In turn, angiotensin-II at 2DIV through these receptors decreases the expression of pro-inflammatory receptors for Interleukin 6 (IL6) and tumor necrosis factor (TNFa). In conclusion, there is a cross-talk between inflammatory factors and Angiotensin-II receptors that should be taken into account when defining an intervention with these drugs.