Wen XiaoNi, Huang YuanGui, Wang JinCun
Department of Neurology, Xijing Hospital, The Fourth Military Medical University of Chinese PLA, China.
Neurol India. 2006 Mar;54(1):58-63; discussion 63. doi: 10.4103/0028-3886.24708.
Selective neuronal loss following status epilepticus (SE) was first described just under 100 years ago. The acute pathology following SE was shown to be 'ischemic cell change' and was assumed to arise through hypoxia/ischemia. Recently, erythropoietin (Epo) has been shown to have potent anti-apoptosis activity in central nervous system neurons in animal models of ischaemic injury.
In this report, in order to determine Epo preconditioning on hippocampus neuronal apoptosis, we examined caspase-3 expression following SE caused by Li-pilocarpine in rats.
Animals were classified into three groups: EP group (pilocarpine group), rhEpo-pilocarpine group and control group. Four hours after preconditioning with Epo intraperitoneally, pilocarpine hydrochloride was administered intraperitoneally and observed for behavioral manifestations of SE. The animals were sacrificed at one hour after SE onset.
At the above-mentioned time point, animals were deeply anesthetized and were perfused through the left ventricle. Detection of hippocampus neuronal apoptosis was performed with caspase-3 immunohistochemical technique on three groups. To further confirm which cell population upregulates caspase-3, brain sections were stained for NeuN (green) and caspase-3 (red).
ANOVA and Fisher's post hoc test was used.
Quatification of hippocampus neurons revealed that the number of caspase-3-positive cells in the CA1/CA3 area and dentate gyrus(DG) of three groups had a significant difference. In comparison with control group, there was an increase by 74% and 534%, 42% and 272% in the CA1/CA3 area and DG of EP group and rhEpo-treated group respectively. There was a decrease by 18% and 26% in the CA1/CA3 area and DG of rhEpo-treated group compared with those in EP group. In addition, colocalization of caspase-3 with NeuN was shown.
Systemic rhEpo therapy reduced caspase-3 expression in SE induced by Li-pilocarpine.
癫痫持续状态（SE）后选择性神经元丢失在不到100年前首次被描述。SE后的急性病理表现为“缺血性细胞改变”，并被认为是由缺氧/缺血引起的。最近，在缺血性损伤动物模型中，已证明促红细胞生成素（Epo）在中枢神经系统神经元中具有强大的抗凋亡活性。
在本报告中，为了确定Epo预处理对海马神经元凋亡的影响，我们检测了大鼠锂-匹罗卡品诱导的SE后caspase-3的表达。
将动物分为三组：EP组（匹罗卡品组）、重组人促红细胞生成素-匹罗卡品组和对照组。腹腔注射Epo预处理4小时后，腹腔注射盐酸匹罗卡品，并观察SE的行为表现。SE发作1小时后处死动物。
在上述时间点，将动物深度麻醉并通过左心室灌注。用caspase-3免疫组化技术对三组动物进行海马神经元凋亡检测。为进一步确定哪些细胞群上调caspase-3，脑切片用NeuN（绿色）和caspase-3（红色）染色。
采用方差分析和Fisher事后检验。
海马神经元定量分析显示，三组动物CA1/CA3区和齿状回（DG）中caspase-3阳性细胞数量有显著差异。与对照组相比，EP组和重组人促红细胞生成素治疗组CA1/CA3区和DG中caspase-3阳性细胞数量分别增加了74%和534%、42%和272%。重组人促红细胞生成素治疗组CA1/CA3区和DG中caspase-3阳性细胞数量与EP组相比分别减少了18%和26%。此外，显示了caspase-3与NeuN的共定位。
全身重组人促红细胞生成素治疗可降低锂-匹罗卡品诱导的SE中caspase-3的表达。
