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[重组人促红细胞生成素预处理对宫内缺氧缺血性脑损伤的保护作用及其对脑组织中半胱天冬酶-3蛋白表达的影响]

[Protection effect of recombiant human erythropoietin preconditioning against intrauterine hypoxic-ischemic brain injury and its influence on expression of caspase-3 protein in brain tissue].

作者信息

Ma Yu-Shan, Zhou Jun, Liu Hui, Du Yu, Lin Xue-Mei

机构信息

Department of Anesthesia, West China Sencond Hospital, Sichuan University, Chendu 610041, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2013 May;44(3):397-401.

Abstract

OBJECTIVE

To investigate the effects of recombine human erythropoietin (rhEPO) on neural cells apoptosis and the expression of Caspase-3 protein in brain tissue of fetal rats after intrauterine hypoxic-ischemic brain injury.

METHODS

Forty-four Sprague-Dawley rats on 19 days of pregnancy were divided into rhEPO treated group, ischemia-reperfusion group and sham-operated group. Intrauterine hypoxic-ischemic injury of fetal rats was induced by bilateral occlusion of the utero-ovarian artery for 20 min. rhEPO (5000 U/kg) was injected into rats through caudal vein in rhEPO treated group while saline was injected into rats in hypoxic-ischemic group 30 min before hypoxic-ischemic injury. The brain samples in rhEPO treated group and hypoxic-ischemic group were obtained at 30 min, 3 h, 6 h, 24 h and 48 h respectively after artery clamping. There was no hypoxic-ischemic injury in sham-operated group, so the brain samples were obtained at 24 hours after sham operation. Neuroapoptosis in brain tissue was measured by TdT mediated dUTP-biotin nick end labeling (Tunel) staining. The expression of Caspase-3 protein was observed by immunohistochemistry.

RESULTS

The number of apoptosis cells in fetal rat hippocampus after intrauterine hypoxic-ischemic increased progressively with reperfusion. Compared with the I/R group, the number of apoptosis cells decreased in rhEPO treated group (P < 0.01). The expression of Caspase-3 increased rapidly after 3 hours from the reperfusion in the I/R group. Compared with the I/R group, there was less expression of Caspase-3 in rhEPO treated group (P < 0.01).

CONCLUSION

rhEPO showed the effects to inhibit the apoptosis of fetal neural cells and the expression of Caspase-3 protein due to intrauterine hypoxic-ischemic brain injury.

摘要

目的

探讨重组人促红细胞生成素(rhEPO)对宫内缺氧缺血性脑损伤胎鼠神经细胞凋亡及脑组织中半胱天冬酶-3(Caspase-3)蛋白表达的影响。

方法

将44只孕19天的Sprague-Dawley大鼠分为rhEPO治疗组、缺血再灌注组和假手术组。通过双侧子宫卵巢动脉闭塞20分钟诱导胎鼠宫内缺氧缺血性损伤。rhEPO治疗组在缺氧缺血损伤前30分钟经尾静脉注射rhEPO(5000 U/kg),而缺氧缺血组注射生理盐水。rhEPO治疗组和缺氧缺血组分别在夹闭动脉后30分钟、3小时、6小时、24小时和48小时获取脑样本。假手术组无缺氧缺血损伤,在假手术后24小时获取脑样本。采用TdT介导的dUTP生物素缺口末端标记(Tunel)染色法检测脑组织中的神经细胞凋亡。通过免疫组织化学观察Caspase-3蛋白的表达。

结果

宫内缺氧缺血后胎鼠海马区凋亡细胞数量随再灌注时间逐渐增加。与缺血再灌注组相比,rhEPO治疗组凋亡细胞数量减少(P < 0.01)。缺血再灌注组再灌注3小时后Caspase-3表达迅速增加。与缺血再灌注组相比,rhEPO治疗组Caspase-3表达较少(P < 0.01)。

结论

rhEPO对宫内缺氧缺血性脑损伤导致的胎鼠神经细胞凋亡及Caspase-3蛋白表达具有抑制作用。

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