[Protection of interleukin-1 receptor antagonist on epileptiform discharges induced by magnesium-free treatment in developing rat cortical neurons].
OBJECTIVE
To study the protection of IL-1ra in cultured developing neuron injury following Mg(2+)-free-induced epileptiform discharges.


METHODS
Rat embryo cortical neurons cultured for 6 d and 17 d were directly exposed to Mg(2+)-free media, or pretreated with IL-1 receptor antagonist or NMDA receptor antagonists before being exposed to Mg(2+)-free media, and then returned to regular media.MTT assay was used to study mitochondrial function injury, laser-scanning confocal microscope to measure [Ca(2+)]i, and real-time RT-PCR to detect gene mRNA expression.


RESULTS
(1) MTT conversion rates were higher in neurons pre and co  treated with 10 mg/L IL-1ra than those of neurons with only Mg(2+)-free treatment in neurons cultured for 17 d, but not in neurons cultured for 6 d.(2) [Ca(2+)]i was lower in neurons pre and co-treated with 10 mg/L IL-1ra than those of neurons with only Mg(2+)-free treatment, either in neurons cultured for 6 d or in neurons cultured for 17 d, and the effects of IL-1ra on [Ca(2+)]i change were different between neurons cultured for 6 d and neurons cultured for 17 d.(3) Pre and co-treated with 10 mg/L IL-1ra NR1 mRNA expression increase induced by Mg(2+)-free treatment was decreased, either in neurons cultured for 6 d or neurons cultured for 17 d, and this effect showed no difference between neurons cultured for 6 d and 17 d; Pre and co-treated with 10 mg/L IL-1ra NR2A mRNA expression increase induced by Mg(2+)-free treatment in neurons cultured for 17 d was decreased, and NR2A mRNA expression showed no difference between IL-1ra group and age-matched control group, but have no effect on neurons cultured for 6 d; Pre and co-treated with 10 mg/L IL-1ra have NR2B mRNA expression increase induced by Mg(2+)-free treatment was not affected, either in neurons cultured for 6 d or neurons cultured for 17 d.


CONCLUSIONS
Neuroprotection of IL-1Ra in seizure-induced injury is age-dependent. The mech-anism of the neuroprotection of IL-1Ra includes down-regulation of [Ca(2+)]i and others.
Department of Pediatrics, Peking University First Hospital, Beijing 100034, China.
Yu-wu
YW
Jiang
Hai-yan
HY
Cao
Jing-min
JM
Wang
Tao
T
Bo
Xi-ru
XR
Wu
D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000196:embryology; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D016291:Dizocilpine Maleate / Q000494:pharmacology; D004827:Epilepsy / Q000378:metabolism / Q000473:pathology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D053590:Interleukin 1 Receptor Antagonist Protein / Q000494:pharmacology*; D008274:Magnesium / Q000494:pharmacology; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology*; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D018123:Receptors, Interleukin / Q000037:antagonists & inhibitors* / Q000502:physiology; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D003470:Culture Media; D018691:Excitatory Amino Acid Antagonists; D053590:Interleukin 1 Receptor Antagonist Protein; D018696:Neuroprotective Agents; D012333:RNA, Messenger; D018123:Receptors, Interleukin; D016194:Receptors, N-Methyl-D-Aspartate; D016291:Dizocilpine Maleate; D008274:Magnesium