外源性重组人促红细胞生成素对胎鼠缺血缺氧性脑损伤后神经细胞凋亡的影响  

Effect of Recombinant Human Erythropoietin on Apoptosis of Neural Cells in Fetal Rats after Intrauterine Hypoxic-ischemic Injury

作　　者：马玉姗[1] 周俊[2] 柳惠[1] 杜宇[1] 林雪梅[1] 

机构地区：[1]四川大学华西第二医院麻醉科,成都610041 [2]四川大学华西口腔医院麻醉科,成都610041

出　　处：《四川大学学报（医学版）》2013年第1期31-35,共5页Journal of Sichuan University(Medical Sciences)

基　　金：四川省科技厅项目(No.2008JY0028-1)资助

摘　　要：目的探讨重组人促红细胞生成素(rhEPO)对胎鼠缺血缺氧性脑损伤后神经细胞凋亡的影响及可能机制。方法将20只孕19d的SD大鼠分为rhEPO治疗组(rhEPO组)、生理盐水缺血对照组(I/R组)和假手术对照组(Sham组)。不同剂量rhEPO治疗组和生理盐水缺血对照组采用钳夹双侧子宫卵巢动脉20min制备宫内缺血缺氧模型,并于宫内缺血前30min分别经尾静脉给予2500、5000、7500U/kg rhEPO或1mL生理盐水,假手术对照组术前30min经尾静脉注射1mL生理盐水后只进行开关腹手术。术后24h取胎鼠观察宫内活胎数,取脑组织,采用免疫组化方法检测活化的Caspase-3蛋白的表达,并通过末端脱氧核苷酸转移酶缺口标记(TUNEL)法观察脑神经元凋亡情况。结果与I/R组比较,Treat组胎鼠死亡率下降(P<0.05)。I/R组海马区可见大量Caspase-3表达阳性细胞。rhEPO组与I/R组比较,Caspase-3蛋白表达明显减弱。rhEPO组中,Caspase-3蛋白的表达随rhEPO剂量的增加呈现减弱趋势,各组间比较差异有统计学意义(P<0.01)。不同剂量rhEPO组细胞凋亡数均较I/R组减少(P<0.01)。各剂量组间比较,2500U/kg组凋亡细胞数高于5000U/kg和7500U/kg组(P<0.01),而5000U/kg与7500U/kg两组之间差异无统计学意义(P>0.05)。结论 rhEPO预处理,可抑制胎鼠宫内缺血缺氧后的神经细胞凋亡,对缺血缺氧性脑损伤具有一定的保护作用。Objective To investigate the effect of recombine human erythropoietin （rhEPO） on apoptosis of neural cells in fetal rats after intrauterine hypoxic-ischemic injury. Methods Twenty SD rats on 19 days of pregnancy were divided into rhEPO （2500 U/kg, 5000 U/kg, 7500 U/kg） treated groups, ischemia-reperfusion （I/ R） group and sham-operated group （4 rats in each group）. Intrauterine hypoxic-ischemic injury of fetal rat was induced by bilateral occlusion of utero-ovarian artery for 20 min. rhEPO was injected into the rats in rhEPO treated group through the caudal vein 30 min before hypoxic-ischemic injury while saline was used in the other two groups. There was no hypoxic-ischemic injury in sham-operated group. The death rate of fetal rats was evaluated at 24 h after the operation, and then the brain samples of fetal rats were harvested. The expression of Caspase-3 protein was observed by immunohistochemistry. Neuroapoptosis was measured by TdT mediated dUTP-biotin nick end labeling （TUNEL） staining. Results Death rates of fetal rats in rhEPO treated groups decreased compared with the I/R group （P〈0.05）. Compared with the I/R group, there was less expression of copious Caspase-3 in rhEPO treated group （P〈0.01）. The expression of Caspase-3 was decreased in the rhEPO treated groups with the increase of rhEPO dose （P〈0.01）. Compared with the I/R group, the death rate of fetal rats in rhEPO treated groups decreased （P〈0.05）, the number of apoptosis cells also decreased obviously （P〈0.01）. The anti-apoptosis effect of 5000 U/kg rhEPO was similar to 7500 U/kg rhEPO, but better than 2500 U/kg rhEPO （P〈0.01）. Conclusion rhEPO can inhibit the apoptosis of fetal rat brain ceils after intrauterine hypoxic-ischemic injury.

关 键 词：重组人促红细胞生成素 缺血缺氧 凋亡 神经保护 

分 类 号：R714.5[医药卫生—妇产科学]