摘要
以Ｆｕｒａ２／ＡＭ为细胞内钙离子的荧光指示剂，用双波长荧光分光光度计测定了缺氧缺糖时体外培养的大鼠胎鼠神经细胞内游离钙（［Ｃａ２＋］ｉ）的变化，并观察了神经生长因子（ＮＧＦ）的影响。结果表明，脑皮质细胞缺氧缺糖培养１６～２４ｈ时，细胞大量死亡。ＮＧＦ剂量依赖地减少神经元缺氧缺糖培养２４ｈ时乳酸脱氢酶（ＬＤＨ）的释放，提高细胞生存力。细胞缺氧缺糖早期引起［Ｃａ２＋］ｉ减少，而后期引起［Ｃａ２＋］ｉ显著升高，导致细胞损害。ＮＧＦ５０μｇ·Ｌ－１在缺氧缺糖早期提高［Ｃａ２＋］ｉ到正常水平，降低后期［Ｃａ２＋］ｉ的升高。提示，ＮＧＦ通过稳定［Ｃａ２＋］ｉ或降低后期的胞内钙升高保护了脑皮质神经元免受缺氧缺糖的损害。
Oxygen/glucose deprived cortical cell cultures were used to explore the involvement of calcium in the mechanism of hypoxic/hypoglycemic insults and protection of nerve growth factor(NGF). Neuron viability and lactate dehydrogenase(LDH) efflux in the bathing medium of cerebral cortex cultures of fetal rats were measured as indication of the NGF effect. Calcium fluorescence indicator Fura2/AM was used to measure free intracellular calcium([Ca2+]i). When cultures were deprived of oxygen/glucose, massive neuronal death occurred 16～24 h following the onset of hypoxia/hypoglycemia. NGF(3～100 μg·L-1) dosedependenly attenuated the 24 h hypoxia/hypoglycemiainduced efflux of LDH and elevated the number of surviving neurons. Hypoxia/hypoglycemia induced a reduction in [Ca2+]i in early stage and then a large elevation of [Ca2+]i in 12～24 h. NGF 50 μg·L-1 brought [Ca2+]i to normal limits during the early stage of hypoxia/hypoglycemia and prevented the later elevation in [Ca2+]i. The [Ca2+]i elevation may be involved in the cell damage in hypoxia/hypoglycemia deprived cultures. It is belived that NGF protect cerebral cortical neurons against hypoxic/hypoglycemic insults via ′stabilizing′ [Ca2+]i level or preventing the late rise in [Ca2+]i.
出处
《药学学报》
CAS
CSCD
北大核心
1998年第5期330-334,共5页
Acta Pharmaceutica Sinica
关键词
脑皮质
神经生长因子
细胞
钙
神经元
缺氧
缺糖
Nerve growth factor(NGF)
Intracellular free Ca2+
Fura2
Neurons
Lactate dehydrogenase
