Map kinase and PKC signaling pathways modulate NGF-mediated apoE transcription

The present study assessed the mechanisms by which nerve growth factor (NGF) increased the level of apolipoprotein E (apoE) in PC12 cells. NGF (50 ng/mL) significantly increased apoE protein levels following 72 h of treatment. Similarly NGF increased luciferase activity in cells transfected with a luciferase reporter construct containing a 500 bp fragment of the apoE promoter, indicating NGF-induced apoE expression is regulated, at least in part, at the level of transcription. The non-selective nitric oxide synthase (NOS) inhibitor Nɷ-nitro-L-arginine methylester (L-NAME; 20 mM) did not attenuate the NGF-mediated increase in luciferase activity, while the inducible NOS inhibitor s-methylisothiourea (S-MIU; 2 mM) partially attenuated this action of NGF. Inhibition of MAP kinase activation with 50 μM U0126 or pre-treatment with the PKC inhibitor bisindolylmaleimide 1 (BIS-1; 10 μM) prevented the NGF-mediated activation of the apoE promoter. Pre-treatment with the phospholipase C (PLC) inhibitor U73122 (5 μM) partially inhibited the NGF-induced increase in luciferase activity while the Akt inhibitor LY294002 (10 μM) had no effect. These data suggest NGF-induced apoE transcription requires MAP kinase and PKC activation and that these TrkA signaling pathways may be modulated by NO.