重组人促红细胞生成素(rhEPO)论文：重组人促红细胞生成素对感染性休克大鼠脑组织保护作用的实验研究.doc

重组人促红细胞生成素(rhEPO)论文：重组人促红细胞生成素对感染性休克大鼠脑组织保护作用的实验研究【中文摘要】重组人促红细胞生成素(recombinant human erythropoietin rhEPO)对感染性休克大鼠脑组织的保护作用。方法健康雄性Vistar大鼠30只,1月龄,随机分为正常对照组(n=6)、感染性休克组(n=12)、rhEPO治疗组(n=12)。感染性休克组及rhEPO治疗组分两个时间点6h和12h,休克组和治疗组以尾静脉注射内毒素(LPS)5mg/kg制备感染性休克模型,治疗组在出现休克症状时立刻腹腔注射EPO5000U/kg,正常对照组大鼠给予等量生理盐水。术后除正常组外各组各时间点大鼠分批腹腔麻醉后以眼球摘除术采血及断头取脑组织,血清行NSE检测,脑组织匀浆行NO检测,提取标本进行病理切片,行HE染色和TUNEL染色检查。每张切片由两位观察者独立计数TUNEL阳性细胞,随机取染色区域5个高倍视野(400)的阳性细胞平均数。结果1、常规HE染色组织病理学显示：正常对照组为正常神经元细胞表现,可见海马区CA1区有3-4层椎体细胞,排列整齐紧密,高倍镜下可见这些神经元核大而圆,透明,有1-2个明显的核仁。而感染性休克组可见海马区椎体细胞变性,核固缩、深染,细胞间隙增大；EPO治疗组海马区椎体细胞存活数量增多,损伤程度均明显减轻。2、TUNEL染色正常组未见TUNEL阳性细胞。感染性休克6小时组可见少量TUNEL阳性细胞；感染性休克12小时组可见大量TUNEL阳性细胞。EPO治疗组阳性细胞数明显减少(P0.01)3、脑组织匀浆NO测定正常组脑组织中NO水平为(2.720.24)mol/t,感染性休克组脑组织中NO水平明显升高,以12小时为重,达(6.350.41)mol/t,明显高于正常组。EPO治疗组脑组织中NO含量明显低于感染性休克组(P0.01)4、血清NSE测定正常组血清NSE水平为(26.262.63)g/ml。感染性休克组血清NSE水平明显升高,以12小时组为重,达(52.423.65)g/ml。EPO治疗组血清NSE含量明显低于感染性休克组(P0.05)结论1、rhEPO能改善模型大鼠的休克症状；2、rhEPO能减少脑组织NO及血清NSE的含量；3、rhEPO能改善脑组织海马区细胞凋亡情况；4、rhEPO对感染性休克大鼠脑细胞有保护作用。【英文摘要】To study the protection role of recombinant human erythropoietin (rhEPO) on brain in the septic shock rats.Method30 rats were randomly divided into three groups:control group(n=6), septic shock group(n=12) and rhEPO group(n=12). The rats of septic shock group and rhEPO group were randomly divided into two time points:6h and 12h, and they were established the models of tail intravenous endotoxin in 5mg/kg doses, in the rhEPO group when the mice appeared shock signs intraperitoneal injection of rhEPO in 5000U/kg doses immediately. And the mice in the control group were only injected amount saline. After surgery all the rats were partial anesthesiaed and eye removal to take the blood to detect the concentration of NSE, then breakaged the mice to get the brain to detect the concentration of NO.6 mice of each group were executed to extract the sample for the pathological section at the different time points, and the sections were stained with Hematoxylin and Eosin and TdT-mediated dUTP nick end labeling.The number of each slice about TUNEL were counted by two independent observer with a average number in 5 random high power fields.Result1、Hematoxylin and Eosin stainHistopathological examination showed:the neurons cells of control group were normal,3-4 layer vertebral somatic closely aligned in hippocampal region, the nuclear of neurons is big round and transparent, having 1-2 prominent nucleoli in high power fields. The vertebral somatic of septic shock were degeneration, nuclear become pyknotic, hyperchromatic and intercellular increase. The survival vertebral somatic in hippocampal region of rhEPO group increasing, damage degree were significantly reduce. 2、TdT-mediated dUTP nick end labeling stainTUNEL-positive cells were not seen in the control group. Many TUNEL-positive cells were seen in the septic shock group with 12h more serious. The TUNEL-positive cells of rhEPO were significantly reduced.3、Brain homogenate NO determinationThe level of NO in brain tissue of the control group was (2.720.24)mol/t, compared with that in the control group, the level of NO in brain tissue of the septic shock group increased significantly with 12h more serious, to (6.350.41)mol/t. The level of NO in brain tissue of the rhEPO group was greatly decreased compared with the septic shock group.4、Serum NSE determinationThe level of NSE of the control group was (26.262.63)g/ml, the level of NSE of the septic shock group increased significantly with 12h more serious, to (52.423.65)g/ml. The level of NSE of the rhEPO group was greatly decreased compared with the septic shock group.Conclusion1、rhEPO can improve shock signs of the model of rats.2、rhEPO can reduce the concentration of NO in brain tissue and NSE of serum.3、rhEPO can reduce apoptosis in neurons of hippocampal region.4、rhEPO have a neuroprotective effect on brain in the septic shock rats.【关键词】重组人促红细胞生成素(rhEPO) 感染性休克脑损伤 NO NSE 细胞凋亡【英文关键词】recombinant human erythropoietin(rhEPO) septic shock brain damage NSE NO cell apoptosis【目录】重组人促红细胞生成素对感染性休克大鼠脑组织保护作用的实验研究摘要3-5ABSTRACT5-6中英文缩略语9-10第1章 前言10-12第2章 材料和方法12-182.1 材料12-132.1.1 手术器械122.1.2 其余物品122.1.3 主要仪器122.1.4 相关仪器12-132.1.5 主要试剂132.2 方法13-182.2.1 动物分组13-142.2.2 感染性休克模型的制作142.2.3 切片制作142.2.4 HE染色14-152.2.5 TUNEL染色15-162.2.6 NO测定16-172.2.7 NSE测定172.2.8 统计学处理17-18第3章 结果18-203.1 常规HE染色183.2 TUNEL染色18-