The in vitro fate of rabbit fetal brain cells after acute in vivo hypoxia
The in vitro fate of rabbit fetal brain cells after acute in vivo hypoxia
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DOI：
10.1523/jneurosci.21-07-j0004.2001
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发表时间：
2001-04-01
影响因子：
5.3
通讯作者：
Tan, S
中科院分区：
文献类型：
--
作者：
Derrick, M;He, J;Tan, S
在缺血性脑损伤的研究中，传统的组织病理学方法估计脑细胞死亡的时间远离缺血性损伤。这些观察结果没有考虑到内源性修复过程或正在进行的损伤级联反应，如凋亡。最初受伤但未被杀死的细胞是最容易被拯救的细胞群。这一假设是体内子宫缺血再灌注将导致体外培养的胎脑细胞中更多的细胞死亡和凋亡。对妊娠29 d的近期妊娠新西兰白色兔进行重复性子宫缺血，缺血40 min，再灌注20 min。子宫缺血后立即取出胎脑，分离成细胞悬液。通过台盼蓝排除法和流式细胞仪上的碘化丙啶（PI）摄取评估，缺血组比非缺血对照组有更多的细胞死亡。将等份的细胞接种并培养24和48小时。缺血组在24小时时比非缺血对照组有更多的细胞死亡（碘化丙啶），并且通过24小时时细胞中的膜联蛋白-V结合和48小时时半胱天冬酶-3活性评估，有更多的凋亡。子宫缺血后，某些胎儿脑细胞立即死亡，其他细胞持续受损，导致坏死和凋亡，这是后来表现出来的。这种方法提供了对这些细胞命运的洞察，并提供了一种评估干预措施以减少细胞损伤的工具。
In the investigation of ischemia-induced brain damage, traditional methods using histopathology estimate brain cell death at a time remote from ischemic insult. These observations fail to take into account endogenous repair processes or ongoing injury cascades like apoptosis. The cells that are injured but not killed initially are the population most amenable to rescue. The hypothesis was that in vivo uterine ischemia-reperfusion would result in more cell death and apoptosis in fetal brain cells cultured in vitro. Near-term, 29 d gestation, pregnant New Zealand White rabbits were subjected to repetitive uterine ischemia for a cumulative time of 40 min ischemia and 20 min reperfusion. Immediately after uterine ischemia, the fetal brains were removed and dissociated into a cell suspension. The ischemic group had more cell death than non-ischemic controls as assessed by Trypan Blue exclusion and propidium iodide (PI) uptake on a flow cytometer. Aliquots of cells were plated and cultured for 24 and 48 hr. The ischemic group had significantly more cell death (propidium iodide) than non-ischemic controls at 24 hr and significantly more apoptosis, as assessed by annexin-V binding in cells at 24 hr and caspase-3 activity at 48 hr. Fewer cells attached to the culture plates at 48 hr in the ischemia group. After uterine ischemia, certain fetal brain cells die immediately, and other cells undergo ongoing damage resulting in necrosis and apoptosis that is manifest later. This method offers insight into the fate of those cells and provides a tool for assessing interventions to decrease cell injury.