Neuroprotective Effect of 2-Methylaminochroman Compound in Human Hippocampal Neuron Cultures

J Korean Neurol Assoc > Volume 14(1); 1996 > Article

Journal of the Korean Neurological Association 1996;14(1): 251-261.

해마 신경원세포의 손상에 대한 2-methylaminochroman 화합물의 보호효과

김명규, 이민철*, 김세종

전남대학교 의과대학 신경과학교실, 병리학교실*

Neuroprotective Effect of 2-Methylaminochroman Compound in Human Hippocampal Neuron Cultures

Myeong Kyu Kim, M.D., Min Cheol Lee*, M.D., Sei Jong Kim, M.D.

Department of Pathology* and Neurology, College of Medicine, Chonnam university

Abstract

It is known that excitotoxicity and oxygen radicals were two major pathogenic events related to mesial temporal sclerosis(MTS), which was the most common histopathologic features in intractable temporal lobe epilepsy. The experiment was designed to investigate the neuroprotective effect of 2-methylaminochroman U-78S17F, a second generation series of nonsteroidal lazaroid compounds, against excitotoxic and oxygen radical injuries on the human fetal hippocampal neurons in vitro. Neuron-enriched cultures were seeded on both 96 well multichamber plates and poly-L-Iysine coated Aclar cover slips to determine cytotoxicity by MTT(3-4, 5-dimethylthiazol-2-yl-2, 5-diphenyl tetrazolium bromide) assay and cytopathologic features respectively. Dose-dependent neuronal injuries were developed by treatment of 100, 200, and 500 microM glutamate (p<0.01) , and 100 microM hypoxanthine plus 10 to 20 mU xanthine oxidase (p<0.01) . The glutamate-induced cytotoxicity was completely blocked by pretreatment of 20 microM MK-801 (p<0.01) , however, U-78S17F did not attenuate the glutamate toxicity. The fetal hippocampal neurons were protected from oxygen radical injuries by pretreatment of 2 to 16 microM U-78517F (p<0.01) . The cytopathologic changes observed by phase-contrast inverted microscope, neurofilament protein (NF) immunocytochemistry, and MTT stain correlated well with the degree of neuronal injuries in experimental groups. Considerably swollen neurons with disintegrated neurites were noted by the excitotoxic and oxygen radical injuries, however, there was no characteristic cytologic difference between them. These data indicated that U-78S17F had only a significant protective effect from oxygen radical injury on fetal hippocampal neurons in culture, and it was suggested that the early treatment of both glutamate-antagonists and antioxidants would be beneficial to reduce MTS following epileptic seizures.