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目的观察重组人红细胞生成素(Recombinant human erythropoietin,rHuEPO)对戊四氮(pentylenetet-razol,PTZ)点燃的癫痫持续状态(status epilepticus,SE)大鼠海马神经元凋亡及磷酸化蛋白激酶B(p-Akt)表达的影响,应用磷脂酰肌醇3激酶(phosphatidylinositol3kinase,PI3K)抑制剂LY294002进一步探讨rHuEPO作用的可能机制。方法采用PTZ点燃SD大鼠制成SE模型,将75只SD大鼠随机分为正常对照组(A组)、PTZ组(B组)、rHuEPO组(C组)、LY294002组(D组)、LY294002溶剂DMSO对照组(E组),每组15只,检测大鼠行为学和脑电图的改变及HE染色观察海马病理学的改变;用TUNEL方法检测海马神经细胞的凋亡情况;免疫组织化学法观察磷酸化蛋白激酶B(P-PKB/p-Akt)的表达。结果A组海马区凋亡细胞数目(7.2±1.0)个与B、C、D、E〔分别为(26.7±3.5)、(16.6±1.1)、(21.1±2.7)和(17.5±1.9)个〕组相比差异均有统计学意义(P<0.01);C、D、E组凋亡细胞数目与B组相比差异均有统计学意义(P<0.05);D组凋亡细胞数目与C、E组比较差异均有统计学意义(P<0.05)。A组p-Akt的阳性细胞数(21.4±1.2)与B、C、D、E组〔分别为(26.6±2.5)、(54.8±5.1)、(33.1±4.9)、(52.4±4.6)〕比较差异均有有统计学意义(P<0.01),且后者在胞核中也开始出现阳性信号;C、D、E组p-Akt的阳性细胞数与B组比较差异均有统计学意义(P<0.05);C、E组p-Akt的阳性细胞数与D组比较差异均有统计学意义(P<0.05)。结论rHuEPO具有抗凋亡、促存活的神经保护作用,PI3K/Akt信号通路可能是rHuEPO发挥神经保护作用的通路之一。其作用机制可能是rHuEPO激活了重要的存活通路PI3K/Akt途径,与提高Akt的活性有关,借以发挥神经保护作用。
Objective To observe the effect of recombinant human erythropoietin (rHuEPO) on the apoptosis of hippocampal neurons in hippocampus of status epilepticus (SE) induced by pentylenetetrazol (PTZ) and the effect of phosphorylated protein kinase B p-Akt) expression, we used phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 to further explore the possible mechanism of rHuEPO. Methods Seventy-five Sprague-Dawley rats were randomly divided into normal control group (group A), PTZ group (group B), rHuEPO group (group C), LY294002 group (group D) LY294002 solvent DMSO control group (E group), 15 rats in each group. The changes of behavior and EEG were detected and the changes of hippocampal pathology were observed by HE staining. The apoptosis of hippocampus neurons was detected by TUNEL method. The expression of phosphorylated protein kinase B (P-PKB / p-Akt) was observed by chemical method. Results The number of apoptotic cells in hippocampus of A group was (26 ± 3.5), (16.6 ± 1.1), (21.1 ± 2.7) and (17.5 ± 1.9), respectively (P <0.01). The number of apoptotic cells in groups C, D and E were significantly different from that in group B (P <0.05). The number of apoptotic cells in group D was significantly correlated with There were significant differences between C and E groups (P <0.05). The number of positive cells of p-Akt in group A (21.4 ± 1.2) and group B, C, D and E [(26.6 ± 2.5), (54.8 ± 5.1), (33.1 ± 4.9) and (52.4 ± 4.6) The difference was statistically significant (P <0.01), and the latter also began to appear in the nucleus positive signal; C, D, E group p-Akt positive cells compared with the B group were statistically significant (P <0.05). The number of p-Akt positive cells in group C and E were significantly different from that in group D (P <0.05). Conclusion rHuEPO has anti-apoptotic and pro-survival neuroprotective effects. PI3K / Akt signaling pathway may be one of the pathways by which rHuEPO exerts neuroprotective effects. Its mechanism may be that rHuEPO activates an important pathway of survival, the PI3K / Akt pathway, which is related to increasing the activity of Akt in order to exert neuroprotective effects.