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样本

资料内容仅供您学习参考，如有不当或者侵权，请联系改正或者删除。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(recombinanthumanerythropoietinrhEPO)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马上腹腔注射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(P<0.01)3、脑组织匀浆NO测定正常组脑组织中NO水平为(2.72±0.24)μmol/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(6.35±0.41)μmol/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(P<0.01)4、血清NSE测定正常组血清NSE水平为(26.26±2.63)μg/ml。感染性休克组血清NSE水平明显升高,以12小时组为重,达(52.42±3.65)μg/ml。EPO治疗组血清NSE含量明显低于感染性休克组(P<0.05)结论1、rhEPO能改进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能改进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】Tostudytheprotectionroleofrecombinanthumanerythropoietin(rhEPO)onbraininthesepticshockrats.Method30ratswererandomlydividedintothreegroups:controlgroup(n=6),septicshockgroup(n=12)andrhEPOgroup(n=12).TheratsofsepticshockgroupandrhEPOgroupwererandomlydividedintotwotimepoints:6hand12h,andtheywereestablishedthemodelsoftailintravenousendotoxinin5mg/kgdoses,intherhEPOgroupwhenthemiceappearedshocksignsintraperitonealinjectionofrhEPOin5000U/kgdosesimmediately.Andthemiceinthecontrolgroupwereonlyinjectedamountsaline.AftersurgeryalltheratswerepartialanesthesiaedandeyeremovaltotakethebloodtodetecttheconcentrationofNSE,thenbreakagedthemicetogetthebraintodetecttheconcentrationofNO.6miceofeachgroupwereexecutedtoextractthesampleforthepathologicalsectionatthedifferenttimepoints,andthesectionswerestainedwithHematoxylinandEosinandTdT-mediateddUTPnickendlabeling.ThenumberofeachsliceaboutTUNELwerecountedbytwoindependentobserverwithaaveragenumberin5randomhighpowerfields.Result1、HematoxylinandEosinstainHistopathologicalexaminationshowed:theneuronscellsofcontrolgroupwerenormal,3-4layervertebralsomaticcloselyalignedinhippocampalregion,thenuclearofneuronsisbigroundandtransparent,having1-2prominentnucleoliinhighpowerfields.Thevertebralsomaticofsepticshockweredegeneration,nuclearbecomepyknotic,hyperchromaticandintercellularincrease.ThesurvivalvertebralsomaticinhippocampalregionofrhEPOgroupincreasing,damagedegreeweresignificantlyreduce.2、TdT-mediateddUTPnickendlabelingstainTUNEL-positivecellswerenotseeninthecontrolgroup.ManyTUNEL-positivecellswereseeninthesepticshockgroupwith12hmoreserious.TheTUNEL-positivecellsofrhEPOweresignificantlyreduced.3、BrainhomogenateNOdeterminationThelevelofNOinbraintissueofthecontrolgroupwas(2.72±0.24)μmol/t,comparedwiththatinthecontrolgroup,thelevelofNOinbraintissueofthesepticshockgroupincreasedsignificantlywith12hmoreserious,to(6.35±0.41)μmol/t.ThelevelofNOinbraintissueoftherhEPOgroupwasgreatlydecreasedcomparedwiththesepticshockgroup.4、SerumNSEdeterminationThelevelofNSEofthecontrolgroupwas(26.26±2.63)μg/ml,thelevelofNSEofthesepticshockgroupincreasedsignificantlywith12hmoreserious,to(52.42±3.65)μg/ml.ThelevelofNSEoftherhEPOgroupwasgreatlydecreasedcomparedwiththesepticshockgroup.Conclusion1、rhEPOcanimproveshocksignsofthemodelofrats.2、rhEPOcanreducetheconcentrationofNOinbraintissueandNSEofserum.3、rhEPOcanreduceapoptosisinneuronsofhippocampalregion.4、rhEPOhaveaneuroprotectiveeffectonbraininthesepticshockrats.【关键词】重组人促红细胞生成素(rhEPO)感染性休克脑损伤NONSE细胞凋亡【英文关键词】recombinanthumanerythropoietin(rhEPO)septicshockbraindamageNSENOcellapoptosis【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

3-5

ABSTRACT

5-6

中英文缩略语

9-10

第1章前言

10-12

第2章材料和方法

12-18

2.1材料

12-13

2.1.1手术器械

12

2.1.2其余物品

12

2.1.3主要仪器

12

2.1.4相关仪器

12-13

2.1.5主要试剂

13

2.2方法

13-18

2.2.1动物分组

13-14

2.2.2感染性休克模型的制作

14

2.2.3切片制作

14

2.2.4HE染色

14-15

2.2.5TUNEL染色

15-16

2.2.6NO测定

16-17

2.2.7NSE测定

17

2.2.8统计学处理

17-18

第3章结果

18-20

3.1常规HE染色

18

3.2TUNEL染色

18-19

3.3脑组织NO含量

19

3.4