Children's National Heart Institute, Children's National Medical Center, Washington, DC 20010, USA.
J Thorac Cardiovasc Surg. 2010 Feb;139(2):489-93. doi: 10.1016/j.jtcvs.2009.05.049. Epub 2009 Oct 23.
Ibuprofen has been shown to reduce cerebral ischemic injury, such as may occur after deep hypothermic circulatory arrest. We investigated whether ibuprofen has direct protective effects against excitotoxic neuronal injury, as may be seen after cerebral ischemia, by using a cell culture model.
Mixed cortical cultures containing neuronal and glial cells were prepared from fetal mice at 13 to 15 days gestation, plated on a layer of confluent astrocytes from 1- to 3-day-old postnatal pups. Near-pure neuronal cultures containing less than 5% astrocytes were obtained from mice of the same gestational stage. Slowly triggered excitotoxic injury was induced at 37 degrees C by 24-hour exposure to 12.5 micromol/L N-methyl-D-aspartate or 50 micromol/L kainate. Neuronal death was quantified by release of lactate dehydrogenase from damaged cells. Data were analyzed using 1-way analysis of variance with Tukey post hoc multiple comparisons.
In mixed cultures, ibuprofen concentrations of 25 microg/mL, 50 microg/mL, and 100 microg/mL all significantly reduced N-methyl-D-aspartate-induced neuronal cell death from 74.5% to 56.1%, 38.7%, and 12.3%, respectively, revealing a strong dose response (P < .001). In near-pure cultures, ibuprofen at a concentration of 25 microg/mL failed to protect neurons, indicating that the neuroprotective effects of ibuprofen require interaction with glial cells. Furthermore, ibuprofen at 100 microg/mL was not protective against neuronal cell death induced by kainate excitotoxicity in near-pure culture but was effective in mixed cultures.
Ibuprofen provides neuroprotection through glial cells against excitotoxic neuronal injury caused by glutamatergic excitotoxicity after cerebral ischemia as demonstrated by reduced neuronal cell death in mixed cell cultures. Further studies are needed to evaluate the potential of ibuprofen to reduce neurologic injury in patients experiencing an hypoxic/ischemic insult.
布洛芬已被证明可减少脑缺血性损伤，例如深低温循环停止后可能发生的损伤。我们通过使用细胞培养模型来研究布洛芬是否对兴奋性毒性神经元损伤具有直接的保护作用，这种损伤可能发生在脑缺血之后。
从孕 13 至 15 天的胎鼠中制备含有神经元和神经胶质细胞的混合皮质培养物，将其铺在由 1 至 3 天龄新生幼鼠的汇合星形胶质细胞层上。从相同孕龄的小鼠中获得含有少于 5%星形胶质细胞的近乎纯神经元培养物。通过在 37°C 下用 12.5μmol/L N-甲基-D-天冬氨酸或 50μmol/L 海人藻酸暴露 24 小时来诱导缓慢触发的兴奋性毒性损伤。通过受损细胞中乳酸脱氢酶的释放来定量神经元死亡。使用单向方差分析和 Tukey 事后多重比较来分析数据。
在混合培养物中，25μg/ml、50μg/ml 和 100μg/ml 的布洛芬浓度分别使 N-甲基-D-天冬氨酸诱导的神经元细胞死亡从 74.5%减少至 56.1%、38.7%和 12.3%，呈现出强烈的剂量反应（P<.001）。在近乎纯的培养物中，25μg/ml 的布洛芬未能保护神经元，表明布洛芬的神经保护作用需要与神经胶质细胞相互作用。此外，在近乎纯的培养物中，100μg/ml 的布洛芬对海人藻酸兴奋性毒性诱导的神经元细胞死亡没有保护作用，但在混合培养物中有效。
布洛芬通过胶质细胞对脑缺血后谷氨酸能兴奋性毒性引起的兴奋性毒性神经元损伤提供神经保护，通过混合细胞培养物减少神经元细胞死亡来证明。需要进一步研究以评估布洛芬降低经历缺氧/缺血损伤的患者神经损伤的潜力。
