백서 피질 신경원 혼합배양 모델에서 Propofol 투여 시점이 허혈-재관류 손상에 미치는 영향: N-Methyl-D-Aspartate 수용체 중심으로 = The influence of propofol administration time on oxygen-glucose deprivation-reperfusion injury in rat mixed cortical cultures focused on N-Methyl-D-Aspartate (NMDA) receptors

다국어 초록 (Multilingual Abstract)

Background: Propofol has been shown to have neuroprotective properties. However, the effect of propofol administration time on neuroprotection is not well understood. This study was conducted to determine if propofol administration time would influence its neuroprotective effects on an in vitro ischemia-reperfusion model, with special attention directed toward NMDA-induced calcium influx.
Methods: Primary mixed cortical cultures of thirteen-day-old rats were exposed to 5 min of oxygen-glucose deprivation (OGD), followed by 2 hr of reperfusion. Propofol (1−100μM) was administered before OGD or administered from the time of OGD to the end of the reperfusion period. In the blank and full kill groups, no drug or NMDA 500μM treatment was given. The surviving cells were counted using trypan-blue staining, and cell death rate (CDR) was determined. To measure the maximum Ca2+ influx, 50μM propofol was pre-treated or co-treated with 100μM NMDA. In the control and NMDA 100μM groups, no drug or NMDA 100μM was given. A P ＜ 0.05 was considered statistically significant.
Results: Cells pre-treated with propofol (10−100μM) or co-treated (50−100μM) at the time of OGD injury had a decreased CDR compared to the blank group. Cells pre-treated (2,713 nA) or co-treated (3,626 nA) with propofol had a decreased maximum Ca2+ influx compared to the 100μM NMDA group (4,075 nA). Cells pre-treated with propofol had a decreased maximum Ca2+ influx compared to co-treated rats.
Conclusions: Propofol demonstrated neuroprotective effects at lower concentrations when administered prior to OGD injury. This may be partially attributable to the reduction of Ca2+ influx against NMDA receptor activation.

다국어 초록 (Multilingual Abstract)

Background: Propofol has been shown to have neuroprotective properties. However, the effect of propofol administration time on neuroprotection is not well understood. This study was conducted to determine if propofol administration time would influence its neuroprotective effects on an in vitro ischemia-reperfusion model, with special attention directed toward NMDA-induced calcium influx.
Methods: Primary mixed cortical cultures of thirteen-day-old rats were exposed to 5 min of oxygen-glucose deprivation (OGD), followed by 2 hr of reperfusion. Propofol (1−100μM) was administered before OGD or administered from the time of OGD to the end of the reperfusion period. In the blank and full kill groups, no drug or NMDA 500μM treatment was given. The surviving cells were counted using trypan-blue staining, and cell death rate (CDR) was determined. To measure the maximum Ca2+ influx, 50μM propofol was pre-treated or co-treated with 100μM NMDA. In the control and NMDA 100μM groups, no drug or NMDA 100μM was given. A P ＜ 0.05 was considered statistically significant.
Results: Cells pre-treated with propofol (10−100μM) or co-treated (50−100μM) at the time of OGD injury had a decreased CDR compared to the blank group. Cells pre-treated (2,713 nA) or co-treated (3,626 nA) with propofol had a decreased maximum Ca2+ influx compared to the 100μM NMDA group (4,075 nA). Cells pre-treated with propofol had a decreased maximum Ca2+ influx compared to co-treated rats.
Conclusions: Propofol demonstrated neuroprotective effects at lower concentrations when administered prior to OGD injury. This may be partially attributable to the reduction of Ca2+ influx against NMDA receptor activation.

참고문헌 (Reference)

• 1 Ligamaneni R, "Widespread inhibition of sodium channel-dependent glutamate release from isolated nerve terminals by isoflurane and propofol" 95 : 1460-1466, 2001
• 2 Abramov AY, "Three distinct mechanisms generate oxygen free radicals in neurons and contribute to cell death during anoxia and reoxygenation" 27 : 1129-1138, 2007
• 3 Choi DW, "The role of glutamate neurotoxicity in hypoxic-ischemic neuronal death" 13 : 171-182, 1990
• 4 Rothman SM, "The neurotoxicity of excitatory amino acids is produced by passive chloride influx" 5 : 1483-1489, 1985
• 5 Kochs E, "The effects of propofol on brain electrical activity, neurologic outcome and neuronal damage following incomplete ischemia in rats" 76 : 245-252, 1992
• 6 Shibuta S, "The effects of propofol on NMDA or nitric oxide mediated neurotoxicity in vitro" 12 : 295-298, 2001
• 7 Zhu H, "The effect of thiopental and propofol on NMDA- and AMPA-mediated glutamate excitotoxicity" 87 : 944-951, 1997
• 8 Murphy PG, "The antioxidant potential of propofol (2,6-diisopropylphenol)" 68 : 613-618, 1992
• 9 Rothman S, "Synaptic release of excitatory amino acid neurotransmitters mediates anoxic neuronal death" 4 : 1884-1891, 1984
• 10 Matsumoto K, "Secondary elevation of extracellular neurotransmitter amino acids in the reperfusion phase following focal cerebral ischemia" 16 : 114-124, 1996
• 11 Li D, "Reperfusion accelerates acute neuronal death induced by simulated ischemia" 206 : 280-287, 2007
• 12 Li C, "Reactive species mechanisms of cellular hypoxia-reoxygenation injury" 282 : 227-241, 2002
• 13 Hans P, "Propofol protects cultured rat hippocampal neurons against N-methyl-D- aspartate receptor-mediated glutamate toxicity" 6 : 249-253, 1994
• 14 Kingston S, "Propofol inhibits phosphorylation of N-methyl-D-aspartate receptor NR1 subunits in neurons" 104 : 763-769, 2006
• 15 Adembri C, "Neuroprotective effects of propofol in models of cerebral ischemia: inhibition of mitochondrial swelling as a possible mechanism" 104 : 80-89, 2006
• 16 Velly LJ, "Neuroprotective effects of propofol in a model of ischemic cortical cell cultures: role of glutamate and its transporters" 99 : 368-375, 2003
• 17 Gunasekar PG, "NMDA receptor activation produces concurrent generation of nitric oxide and reactive oxygen species: implications for cell death" 65 : 2016-2021, 1995
• 18 Goldberg MP, "N-methyl-D- aspartate receptors mediate hypoxic neuronal injury in cortical culture" 243 : 784-791, 1987
• 19 Fujimoto S, "Mechanisms of oxygen glucose deprivation-induced glutamate release from cerebrocortical slice cultures" 50 : 179-187, 2004
• 20 Roettger V, "Mechanism of glutamate release from rat hippocampal slices during in vitro ischemia" 75 : 677-685, 1996
• 21 Orser BA, "Inhibition by propofol (2,6 di-isopropylphenol) of the N-methyl-D-aspartate subtype of glutamate receptor in cultured hippocampal neurones" 116 : 1761-1768, 1995
• 22 Phillis JW, "Inhibition by anion channel blockers of ischemia-evoked release of excitotoxic and other amino acids from rat cerebral cortex" 758 : 9-16, 1997
• 23 Calò G, "Immediate and delayed effects of in vitro ischemia on glutamate efflux from guinea-pig cerebral cortex slices" 751 : 300-306, 1997
• 24 Rossi DJ, "Glutamate release in severe brain ischaemia is mainly by reversed uptake" 403 : 316-321, 2000
• 25 Rothman SM, "Glutamate and the pathophysiology of hypoxic ischemic brain damage" 19 : 105-111, 1986
• 26 Choi DW, "Excitotoxic cell death" 23 : 1261-1276, 1992
• 27 Benveniste H, "Elevation of the extracellular concentrations of glutamate and aspartate in rat hippocampus during transient cerebral ischemia monitored by intracerebral microdialysis" 43 : 1369-1374, 1984
• 28 Yamakura T, "Effects of propofol on various AMPA-, kainate- and NMDA-selective glutamate receptor channels expressed in Xenopus oocytes" 188 : 187-190, 1995
• 29 Grasshoff C, "Effects of propofol on N-methyl-D- aspartate receptor-mediated calcium increase in cultured rat cerebrocortical neurons" 22 : 467-470, 2005
• 30 Amakawa K, "Effects of pre- and postischemic administration of thiopental on transmitter amino acid release and histologic outcome in gerbils" 85 : 1422-1430, 1996
• 31 Phillis JW, "Characterization of modes of release of amino acids in the ischemic/reperfused rat cerebral cortex" 43 : 461-467, 2003
• 32 Drejer J, "Cellular origin of ischemia-induced glutamate release from brain tissue in vivo and in vitro" 45 : 145-151, 1985
• 33 Katayama Y, "Calcium-dependent glutamate release concomitant with massive potassium flux during cerebral ischemia in vivo" 558 : 136-140, 1991
• 34 Tabor MP, "An advanced in vitro model to study hypoxia/low glucose-induced neuronal cell damage and death" 825 : 267-278, 1997