重组人红细胞生成素对癫痫持续状态大鼠海马神经元磷酸化Akt表达的影响

Effect of rHuEPO on the expression of p-Akt in hippocampus of statural epilepticus rats

目的观察重组人红细胞生成素对戊四氮所致的癫痫持续状态(statusepil epticus,SE)的SD大鼠海马神经元磷酸化Akt(p-Akt)表达的影响,并应用PI3K抑制剂LY294002进行干预,进一步探讨重组人红细胞生成素是否对SE大鼠的海马神经细胞具有保护作用及作用的可能机制。方法采用戊四氮点燃大鼠SE模型。75只SD大鼠随机分为5组(n=15),A组:正常对照组(腹腔注射生理盐水);B组:生理盐水治疗组(腹腔注射戊四氮点燃SE发作后30分钟,再腹腔注射生理盐水);C组:重组人红细胞生成素治疗组(腹腔注射戊四氮点燃SE发作后30分钟,再腹腔注射重组人红细胞生成素);D组:PI3K抑制剂LY294002干预组(腹腔注射戊四氮点燃SE发作后10分钟,再脑室内注射LY294002,SE发作后30分钟腹腔注射重组人红细胞生成素);E组:DMSO(二甲基亚砜)对照组(腹腔注射戊四氮点燃SE发作后10分钟,再脑室注射LY294002的溶剂DMSO,SE发作后30分钟腹腔注射重组人红细胞生成素),检测各组大鼠行为、脑电图的改变及HE染色观察各组海马病理学的改变、免疫组织化学法观察各组磷酸化Akt的表达。结果重组人红细胞生成素治疗组较生理盐水治疗组磷酸化Akt的表达增多;PI3K抑制剂LY294002干预组海马磷酸化Akt表达较重组人红细胞生成素治疗组减少。结论重组人红细胞生成素对SE大鼠的海马神经元具有保护作用,其作用机制可能是重组人红细胞生成素提高了Akt的活性,激活了存活通路PI3K/Akt途径,这可能是重组人红细胞生成素发挥神经保护作用的机制之一。

Objective To observe the effect of recombinant Human erythropoietin（rHuEPO） on the expression of p-Akt in hippocampus of statural epilepticus（SE） rats kindled by the Pentylenetetrazol（PTZ） and the influence of LY294002 which is the inhibitor phosphatidyl inositol 3 kinase, to explore the possible neuroprotective mechanism of rHuEPO to the SE rats. Methods SE model kindled by Pentylenetetrazol .The 75 SD rats were divided into normal control group（normal saline intraperitoneal injection, group A）、saline treated group（normal saline intraperitoneal injection 30 minutes after the SE kindled by the PTZ, group B）、rHuEPO treated group （rHuEPO intraperitoneal injection 30 minutes after the SE kindled by the PTZ, group C）、 LY294002 intervention group （LY294002 intraventricular injection 10 minutes and rHuEPO intraperitoneal injection 30 minutes after the SE kindled by the PTZ respectively, group D）、DMSO control group（DMSO intraventricular injection 10 minutes and rHuEPO intraperitoneal injection 30 minutes after the SE kindled by the PTZ respectively, group E）.Changes of behavior、EEG recording and hippocampal histology were observed; the expression of p-Akt were detected through immunohistochemical method. Results The expression of p-Akt was increased in rHuEPO treated group than in saline treated group. The expression of p-Akt was decreased obviously in LY294002 intervention group comparied with group rHuEPO. Conclusion The rHuEPO showed the effect of neuroprotection in SE rats, the pathway of PI3K/Akt may be one of the neuroprotective ways of rHuEPO. The possible mechanism is rHuEPO activated the PI3K/Akt pathway and then up-regulated the activity of Akt to displayed neuroprotective effect.