Effect of Trolox, Cycloheximide or MK-801 on the Neuronal Cell Death Induced by FeCl2, Buthionine Sulfoximine or KCN in Primary Murine Mixed Cortical Cell Culture.
- Author:
In Hwan KIM
1
;
Jung Gil LEE
;
Tae Sun KIM
;
Shin JUNG
;
Jae Hyoo KIM
;
Soo Han KIM
;
Sam Suk KANG
;
Je Hyuk LEE
;
Kuy Sook LEE
;
Jong Keun KIM
Author Information
1. Department of Neurosurgery, Chonnam University, Medical School, Kwangju, Korea.
- Publication Type:Original Article ; In Vitro
- Keywords:
Oxidative stress;
Murine cerebral cortical cell culture;
Neuronal death
- MeSH:
Adenosine Triphosphate;
Baths;
Buthionine Sulfoximine*;
Cell Culture Techniques*;
Cell Death*;
Cell Membrane;
Cycloheximide*;
Dizocilpine Maleate*;
Epilepsy;
Glutathione;
Hand;
Lipid Peroxidation;
Neurodegenerative Diseases;
Neurons*;
Oxidative Stress;
Potassium;
Stroke;
Vitamin E;
Vitamins;
Water
- From:Journal of Korean Neurosurgical Society
1997;26(10):1342-1350
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
Oxidative stress is known to be a major neuropathologic mechanism in chronic neurodegenerative disorders as well as in stroke, trauma and epilepsy, and many kinds of oxidative insults induce neuronal injury. The purpose of this study was to examine the temporal effects of trolox(TLX; water and lipid soluble vitamin E analog), cycloheximide(CHX; protein synthesis inhibitor) and MK-801(NMDA receptor antagonist) on neuronal death indu-ced by different kinds of oxidative insults in primary murine mixed cortical cell culture(14-16 days in vitro), and to gain information on the mechanisms underlying oxidative cell death. As oxidative insults, the authors used iron(FeCl2), buth-ionine sulfoximine(BSO; glutathione depletor) and potassium cyanide(KCN; ATP depletor). Cell death was assessed by measurement of LDH efflux to bathing media at the end of exposure. All three agents induced neuronal cell death associated with cell body swelling. FeCl2(30nM-1mM) induced conce-ntration- and exposure time-dependent neurotoxicity, while BSO(10nM-3mM) showed little neurotoxicity at the end of 24 hrs exposure, but marked neuronal cell death at the end of 48 hrs; at concentrations of over 100uM of BSO neurotoxicity reached a plateau. KCN(0.1mM-1mM) also showed dose-dependent neurotoxicity. TLX(100nM) did not affect the neurotoxicity induced by KCN(1mM) but almost completely block BSO(1mM)- or FeCl2(100, 300nM)- induced neuronal cell death. CHX(1ng/ml) significantly attenuated BSO-induced cell death but did not protect against KCN(1mM)-induced cell death. CHX treatment, on the other hand, significantly potentiated FeCl2(100 or 300nM)-induced death. MK-801(10nM) markedly inhibited KCN-induced cell death but had no effect on FeCl2-induced death. MK-801 also significantly attenuated BSO-induced neurotoxicty after exposure for 48hrs but this protective effect disappeared at the end of 72hr. These results suggest that protein synthesis as well as lipid peroxidation of cell membrane may involve oxidative neuronal injury and that one oxidative agent may induce various cell death processes.
