시험관내허혈환경에서과립구성장인자의인간뇌피질에서유래된신경세포주의보호효과

Abstract

Stroke & Neural Stem cell Laboratory in Clinical Research Institute, Department of Neurology, Seoul National University Hospital, Seoul National University, Seoul, South Korea*,Department of Neurology, Seoul National Hospital, Seoul, South Korea†Department of Neurology, Ilsan Paik Hospital, Inje University, Goyang, South Korea‡ Background: Granulocyte colony-stimulating factor (G-CSF) has neuroprotective effects against experimentalfocal cerebral ischemia. However, its neuroprotective effect on human brain is unknown. In this study we attempt-ed to determine whether G-CSF can protect the neuronal cell line derived from human cortex against in vitroischemia.Methods: Human cortical-neuroblastoma hybrid cell line (A1G11) was exposed to hypoxia with glu-cose deprivation (6 hours). To test the protective effect of G-CSF, human recombinant G-CSF (0, 5, 25, 50, or 100ng/ml) was used. Cell viability or cytotoxicity was determined by MTT or LDH assay. Flow cytometry by annex-in-V and propidium iodide (PI) and immunocytochemistry by active caspase-3 antibody were performed to detectapoptotic or viable cells. Results: Morphological changes suggestive of neuronal damage were observed follow-ing ischemia. G-CSF promoted cell survival (42% vs 32%) and decreased cytotoxicity (9% vs 18%) at a maximal-ly effective concentration of 25 ng/ml. From the flow cytometric analysis, G-CSF reduced early apoptotic (annex-in V+/PI-), and late apoptotic or necrotic (annexin V+/PI+) cells. A decrease in immunoreactivity against activecaspase-3 in the G-CSF-treated group was correlated with the neuronal protection.Conclusions: Our results pro-vide evidence that G-CSF can protect human cortical hybrid neurons from apoptotic or necrotic cell death after anischemic insult. 대한뇌졸중학회지제6권제1호2004년5월