Korean J Perinatol.  2004 Mar;15(1):34-40.

Neuroprotective Effects of MK-801 (Dizocilpine) on Bilirubin Neurotoxicity in Mouse Cortical Cell Culture

• Hwang JH
• Choi CW
• Chang YS
• Park WS

Abstract

OBJECTIVE
To evaluate whether MK-801, N-methyl-D-aspartate (NMDA) receptor antagonist, could protect against bilirubin neurotoxicity in mouse neuronal cells.
METHODS
Mouse cerebral cortical cells were obtained from 14 day-old mouse fetal cerebral cortex primary culture. Cerebral cortical cells were exposed to medium containing concentrations of bilirubin 100 microM and additionally treated with 5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohept-5,10-imine (MK-801, dizocilpine), 1,2,3,4-tetrahydroxy-6-nitro-2,3-dioxo-benzo [f] quinoxaline-7-sulfonamide disodium (NBQX), NG-nitro-L-arginine methyl ester hydrochloride (L-NAME) for 4 hours. Then, the bilirubin cytotoxicity for cerebral cortical cells was quantitated by the activity of LDH released from cerebral cortical cells into the culture media. Cortical cells were stained with propidium iodide and visualized by confocal laser scanning microscopy for detection of apoptotic cells.
RESULTS
After exposure for 4 hours, the cytotoxicity for cortical cells was the least in bilirubin treated with MK-801 compared to bilirubin alone, bilirubin treated with NBQX, and L-NAME (32+/-9%, 55+/-7%, 52+/-8%, 53+/-9%, respectively, p<0.05). Cells treated with MK-801 had fewer apoptotic nuclei evident on confocal microscopy.
CONCLUSION
Our study suggests bilirubin neurotoxicity is mediated NMDA receptors and MK-801 is capable to prevent bilirubin-induced neuronal damage.

Keyword

Bilirubin neurotoxicity; NMDA receptor; MK-801; NBQX; L-NAME

MeSH Terms

Animals
Bilirubin*
Cell Culture Techniques*
Cerebral Cortex
Culture Media
Dizocilpine Maleate*
Mice*
Microscopy, Confocal
N-Methylaspartate
Neurons
Neuroprotective Agents*
NG-Nitroarginine Methyl Ester
Propidium
Receptors, N-Methyl-D-Aspartate
Bilirubin
Culture Media
Dizocilpine Maleate
N-Methylaspartate
NG-Nitroarginine Methyl Ester
Neuroprotective Agents
Propidium
Receptors, N-Methyl-D-Aspartate