Poststress treatment with PNU282987 can rescue SH-SY5Y cells undergoing apoptosis via α7 nicotinic receptors linked to a Jak2/Akt/HO-1 signaling pathway.
Most neuroprotection studies with nicotinic agonists have shown efficacy when given before the stressor. Here we have investigated whether the α7 nicotinic acetylcholine receptor (nAChR) agonist PNU282987 can prevent cell death once the cells have already undergone an oxidative stress. The combination of rotenone (30 μM) plus oligomycin A (10 μM) (rot/oligo) has been used as an in vitro model of mitochondrial ROS production. SH-SY5Y cells incubated with rot/oligo for 8h and left for another 16 h in MEM/F-12 experienced 30% apoptotic cell death. Under these experimental conditions, PNU282987 administered after rot/oligo (PST/PNU) prevented cell death in a concentration-dependent manner. Co-incubation of PNU282987 with 100 nM methyllycaconitine (a selective α7 nAChR antagonist), 10 μM mecamylamine (a nonselective nAChR antagonist), 3 μM LY294002 (a PI3K inhibitor), or 10 μM AG490 (a Jak2 inhibitor) prevented the protection afforded by PST/PNU. Moreover, the increase in ROS, active caspase-3, and apoptosis caused by rot/oligo was also prevented by PST/PNU. Furthermore, PNU282987 increased the expression of heme oxygenase-1 (HO-1), a critical cell defense enzyme against oxidative stress; this increase was prevented by AG490 or LY294002. The HO-1 inhibitor Sn(IV) protoporphyrin-IX also inhibited the PST/PNU protecting effects. These results suggest that activation of α7 nAChR linked to the Jak2/PI3K/Akt cascade induces the antioxidant enzyme HO-1 to provide neuroprotection.
Parada
Esther
E
Instituto Teófilo Hernando, Facultad de Medicina, Universidad Autónoma de Madrid, Madrid, Spain.
Egea
Javier
J
Romero
Alejandro
A
del Barrio
Laura
L
García
Antonio G
AG
López
Manuela G
MG
D000157:Aconitine / Q000031:analogs & derivatives / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D001549:Benzamides / Q000494:pharmacology*; D001643:Bridged Bicyclo Compounds / Q000494:pharmacology*; D002478:Cells, Cultured; D019610:Cytoprotection / Q000187:drug effects; D004353:Drug Evaluation, Preclinical; D051547:Heme Oxygenase-1 / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D053614:Janus Kinase 2 / Q000378:metabolism / Q000502:physiology*; D018722:Nicotinic Agonists / Q000494:pharmacology; D018733:Nicotinic Antagonists / Q000494:pharmacology; D051058:Oncogene Protein v-akt / Q000378:metabolism / Q000502:physiology*; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D011978:Receptors, Nicotinic / Q000378:metabolism / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013997:Time Factors; D064569:alpha7 Nicotinic Acetylcholine Receptor
D001549:Benzamides; D001643:Bridged Bicyclo Compounds; C579167:Chrna7 protein, human; D018722:Nicotinic Agonists; D018733:Nicotinic Antagonists; C498513:PNU-282987; D017382:Reactive Oxygen Species; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; C054634:methyllycaconitine; C496541:HMOX1 protein, human; D051547:Heme Oxygenase-1; C507924:JAK2 protein, human; D053614:Janus Kinase 2; D051058:Oncogene Protein v-akt; D000157:Aconitine