中华物理医学与康复杂志
中華物理醫學與康複雜誌
중화물리의학여강복잡지
CHINESE JOURNAL OF PHYSICAL MEDICINE AND REHABILITATION
2008年
3期
156-160
,共5页
大鼠%缺血再灌注%巢蛋白%红细胞生成素
大鼠%缺血再灌註%巢蛋白%紅細胞生成素
대서%결혈재관주%소단백%홍세포생성소
Rat%Ischemia-reperfusion%EPO%Nestin
目的 本实验以巢蛋白(nestin)作为神经干细胞标志物,应用大鼠全脑缺血模型研究Nestin与红细胞生成素(EPO)的表达变化规律及其关系,初步探讨内源性神经干细胞增殖、分化的相关机制.方法 共选取健康成年SD大鼠36只,将其分为正常对照组及实验组.实验组采用Pulsinelli四血管闭塞法制作大鼠全脑缺血模型,于全脑缺血30 min再灌注3 h、6 h、12 h、24 h、3 d、7 d、14 d及21 d时各取4只大鼠处死取材,将脑组织标本制作成石蜡切片,应用免疫组织化学染色法检测Nestin与EPO表达水平.结果 (1)正常对照组海马区、室旁区、皮质区均未见Nestin染色阳性细胞,实验组缺血再灌注3 h室管膜下区、海马区即有Nestin染色阳性细胞表达,于再灌注14 d时达到高峰,再灌注21 d仍有表达,但表达强度逐渐减弱.对照组与实验组各时间段Nestin均数进行方差分析,发现差异具有统计学意义(P<0.05),实验组各相邻时间段Nestin均数间相比,差异均具有统计学意义(均P<0.05);(2)正常对照组大鼠海马区可见少量EPO表达,实验组缺血再灌注3 h时开始有少量表达,于再灌注24 h时达到高峰,以后则逐渐减弱,但在缺血21 d后仍有表达.正常对照组与实验组各时间段EPO均数经方差分析,发现差异具有统计学意义(P<0.05);实验组EPO表达水平除再灌注7 d与3 d时差异无统计学意义(P>0.05)外,其余各相邻均数间差异均有统计学意义(均P<0.05).结论 大鼠全脑缺血再灌注后脑组织Nestin、EPO阳性表达细胞增加;EPO阳性细胞表达增加是脑组织神经损伤后早期保护性反应之一;Nestin、EPO的表达规律具有一致性,EPO可能具有促进神经干细胞增殖的功能.
目的 本實驗以巢蛋白(nestin)作為神經榦細胞標誌物,應用大鼠全腦缺血模型研究Nestin與紅細胞生成素(EPO)的錶達變化規律及其關繫,初步探討內源性神經榦細胞增殖、分化的相關機製.方法 共選取健康成年SD大鼠36隻,將其分為正常對照組及實驗組.實驗組採用Pulsinelli四血管閉塞法製作大鼠全腦缺血模型,于全腦缺血30 min再灌註3 h、6 h、12 h、24 h、3 d、7 d、14 d及21 d時各取4隻大鼠處死取材,將腦組織標本製作成石蠟切片,應用免疫組織化學染色法檢測Nestin與EPO錶達水平.結果 (1)正常對照組海馬區、室徬區、皮質區均未見Nestin染色暘性細胞,實驗組缺血再灌註3 h室管膜下區、海馬區即有Nestin染色暘性細胞錶達,于再灌註14 d時達到高峰,再灌註21 d仍有錶達,但錶達彊度逐漸減弱.對照組與實驗組各時間段Nestin均數進行方差分析,髮現差異具有統計學意義(P<0.05),實驗組各相鄰時間段Nestin均數間相比,差異均具有統計學意義(均P<0.05);(2)正常對照組大鼠海馬區可見少量EPO錶達,實驗組缺血再灌註3 h時開始有少量錶達,于再灌註24 h時達到高峰,以後則逐漸減弱,但在缺血21 d後仍有錶達.正常對照組與實驗組各時間段EPO均數經方差分析,髮現差異具有統計學意義(P<0.05);實驗組EPO錶達水平除再灌註7 d與3 d時差異無統計學意義(P>0.05)外,其餘各相鄰均數間差異均有統計學意義(均P<0.05).結論 大鼠全腦缺血再灌註後腦組織Nestin、EPO暘性錶達細胞增加;EPO暘性細胞錶達增加是腦組織神經損傷後早期保護性反應之一;Nestin、EPO的錶達規律具有一緻性,EPO可能具有促進神經榦細胞增殖的功能.
목적 본실험이소단백(nestin)작위신경간세포표지물,응용대서전뇌결혈모형연구Nestin여홍세포생성소(EPO)적표체변화규률급기관계,초보탐토내원성신경간세포증식、분화적상관궤제.방법 공선취건강성년SD대서36지,장기분위정상대조조급실험조.실험조채용Pulsinelli사혈관폐새법제작대서전뇌결혈모형,우전뇌결혈30 min재관주3 h、6 h、12 h、24 h、3 d、7 d、14 d급21 d시각취4지대서처사취재,장뇌조직표본제작성석사절편,응용면역조직화학염색법검측Nestin여EPO표체수평.결과 (1)정상대조조해마구、실방구、피질구균미견Nestin염색양성세포,실험조결혈재관주3 h실관막하구、해마구즉유Nestin염색양성세포표체,우재관주14 d시체도고봉,재관주21 d잉유표체,단표체강도축점감약.대조조여실험조각시간단Nestin균수진행방차분석,발현차이구유통계학의의(P<0.05),실험조각상린시간단Nestin균수간상비,차이균구유통계학의의(균P<0.05);(2)정상대조조대서해마구가견소량EPO표체,실험조결혈재관주3 h시개시유소량표체,우재관주24 h시체도고봉,이후칙축점감약,단재결혈21 d후잉유표체.정상대조조여실험조각시간단EPO균수경방차분석,발현차이구유통계학의의(P<0.05);실험조EPO표체수평제재관주7 d여3 d시차이무통계학의의(P>0.05)외,기여각상린균수간차이균유통계학의의(균P<0.05).결론 대서전뇌결혈재관주후뇌조직Nestin、EPO양성표체세포증가;EPO양성세포표체증가시뇌조직신경손상후조기보호성반응지일;Nestin、EPO적표체규률구유일치성,EPO가능구유촉진신경간세포증식적공능.
Objective To observe dynamic changes of Nestin and Erythropoietin(EPO)expression after cerebral ischemia and investigate possible mechanism of intrinsic neurogenesis. Methods A total of 36 SpragueDawley rats were divided into a control group(n=4)and an experimental group(n=32)randomly.The experimental group were further divided into 8 subgroups corresponding to the observation time points of 3,6,12,24 hours and 3,7,14,and 21 days after reperfusion.The model of experimental ischemia was made by 4-VO.The specimens were made into paraffin section.The expression of Nestin and EPO were detected by immunohistochemistry method. Results (1)Nestin detection:No Nestin positive cells were observed in hippocampal zone,subventricular zone (SVZ)and contex in the control group.The expression of Nestin started at 3 h in SVZ but not in hippocampal zone and it started to increase at 6 h after cerebral ischemia and reach a peak at 14d after cerebral isehemia in the hippocampal zone,then decreased at 21 d.The difference of Nestin expression among different time points is statistically significant(P<0.05).(2)EPO detection:A few EPO positive cells were observed in hippocampal zone in the control group.The expression of EPO started to increase at 3 h after cerebral ischemia and reach a peak at 24 h after cerebral ischemia,then decreased with time.The difference of EPO expression among the different time points is statistically significant(P<0.05). Conclusion (1)The increased expression of EPO and Nestin after cerebral ischemia might be a beneficial protective response of cells to the ischemic injury.(2)The sequence of expression of EPO and Nestin after cerebral ischemia is relevant.EPO may promote proliferation of NSC.
