Our lab has reported that overexpression of angiotensin converting enzyme 2 (ACE2) in the brain blunts the development of angiotensin‐II (Ang‐II)‐induced hypertension and sympathetic activation. Yet, the signaling mechanisms are still elusive. Hypothesis: Ang‐II‐mediated phosphorylation of MAP kinase and Akt in neurons is attenuated by ACE2 overexpression. A Neuro‐2a cell line overexpressing ACE2 was generated by stable transfection with an expression plasmid for human ACE2 (N2AhACE2). Control and N2AhACE2 cells were treated with 100 nM Ang‐II or vehicle, in presence and absence of inhibitors. Phosphorylation of p38 MAP kinase (Thr180/Thr182) and Akt (Ser473) by Ang‐II in control cells (2‐ and 1.5‐fold increase, respectively, p<0.05) (Western blot), was prevented by ACE2 overexpression. Pretreatment with DX600 (ACE2 inhibitor) for 30 min had no effect in control cells, but restored Ang‐II‐induced phosphorylation in N2AhACE2. In addition, Ang‐II‐induced expression of AT1R (1.8‐fold increase vs. vehicle, p<0.05) was blunted by Losartan (AT1R antagonist), or ACE2 overexpression. Furthermore, pretreatment with A‐779 (Ang‐(1–7) receptor antagonist) restored phosphorylation in N2AhACE2. These data suggest that ACE2 counteracts Ang‐II‐induced phosphorylation of p38 MAP kinase and Akt, by both, decreasing AT1R signaling and increasing Ang‐(1–7) receptor signaling in neurons. (Support: HL093178)
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