Estrogen receptor agonists and estrogen attenuate TNF-α-induced apoptosis in VSC4.1 motoneurons.
Tumor necrosis factor-alpha (TNF-α) may cause apoptosis and inflammation in amyotrophic lateral sclerosis (ALS) and spinal cord injury (SCI). Recent studies suggest that estrogen (EST) provides neuroprotection against SCI. We tested whether 1,3,5-tris (4-hydroxyphenyl)-4-propyl-1H-pyrazole (PPT) (EST receptor alpha (ERα) agonist), 2,3-bis (4-hydroxyphenyl) propionitrile (DPN) (EST receptor beta (ERβ) agonist), or EST itself would prevent apoptosis in VSC4.1 motoneurons following exposure to TNF-α. Cells were exposed to TNF-α and 15 min later treated with PPT, DPN, or EST. Posttreatment with 50 nM PPT, 50 nM DPN, or 150 nM EST prevented cell death in VSC4.1 motoneurons. Treatment of VSC4.1 motoneurons with PPT, DPN, or EST induced overexpression of ERα, ERβ, or both, which contributed to neuroprotection by upregulating expression of anti-apoptotic proteins (p-AKT, p-CREB, Bcl-2, and p-Src). Our analyses also revealed that EST agonists and EST increased phosphorylation of extracellular signal-regulated kinase (ERK). The L-type Ca(2+) channel inhibitor, nifedipine (10 μM), partially inhibited EST agonist and EST-induced increase in phosphorylated ERK expression. The mitogen-activated protein kinase inhibitor, PD98059 (5 μM), partially prevented ER agonists and EST from providing neuroprotection to TNF-α toxicity. Presence of the nuclear ER antagonist, ICI 182 780 (10 μM), blocked the neuroprotection provided by all three ER agonists tested. Taken together, our data indicate that both ERα and ERβ contribute to PPT, DPN, or EST-mediated neuroprotection with similar signaling profiles. Our data strongly imply that PPT, DPN, or EST can be used as effective neuroprotective agents to attenuate motoneuron death in ALS and SCI.
Department of Neurosciences, Medical University of South Carolina, 96 Jonathan Lucas Street, Charleston, South Carolina 29425, USA.
Arabinda
A
Das
Joshua A
JA
Smith
Cameron
C
Gibson
Abhay K
AK
Varma
Swapan K
SK
Ray
Naren L
NL
Banik
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015415:Biomarkers / Q000378:metabolism; D020746:Calcium Channels, L-Type / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002459:Cell Fusion; D002460:Cell Line; D045744:Cell Line, Tumor; D004622:Embryo, Mammalian / Q000166:cytology; D004967:Estrogens / Q000494:pharmacology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D006822:Hybrid Cells / Q000378:metabolism; D009046:Motor Neurons / Q000187:drug effects*; D009447:Neuroblastoma / Q000473:pathology; D018696:Neuroprotective Agents / Q000494:pharmacology; D009570:Nitriles / Q000494:pharmacology; D010636:Phenols / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D011720:Pyrazoles / Q000494:pharmacology; D051381:Rats; D011960:Receptors, Estrogen / Q000819:agonists*; D015398:Signal Transduction / Q000187:drug effects; D013116:Spinal Cord / Q000196:embryology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D011422:Propionates
D015415:Biomarkers; D020746:Calcium Channels, L-Type; D004967:Estrogens; D048049:Extracellular Signal-Regulated MAP Kinases; D018696:Neuroprotective Agents; D009570:Nitriles; D010636:Phenols; D011720:Pyrazoles; D011960:Receptors, Estrogen; D014409:Tumor Necrosis Factor-alpha; C486182:2,3-bis(4-hydroxyphenyl)-propionitrile; D011422:Propionates; C486184:4,4',4''-(4-propyl-((1)H)-pyrazole-1,3,5-triyl) tris-phenol