大鼠脑缺血再灌注后大脑皮质细胞凋亡的体视学观察

发布时间：2018-06-22 22:51

  本文选题：脑缺血再灌注 + 凋亡　； 参考：《南昌大学》2008年硕士论文

【摘要】： 为了观察脑缺血再灌注后不同存活时间内大鼠大脑皮质不同区域细胞凋亡的变化情况,本实验选取42只SD大鼠,将其随机分为对照组,缺血15min再灌注6h、12h、24h组,缺血2h再灌注6h、12h、24h组,每组6只。通过结扎左侧颈总动脉,动脉夹夹闭右侧颈总动脉15min和2h后松开动脉夹恢复血流的方法构建脑缺血再灌注模型。采用Hoechst33258荧光染色和Bax免疫组化方法观察海马CA1、CA2、CA3、CA4区、齿状回及顶叶皮质区细胞凋亡情况,用体视学参数——体积密度(Vv)和数密度(Nv)定量地分析这些区域细胞在脑缺血再灌注后不同存活时间内的凋亡情况。结果如下: Hoechst33258荧光染色:凋亡细胞细胞核在紫外光下呈现出明亮鲜艳的蓝色荧光。各缺血再灌注组中上述大脑皮质各部位凋亡细胞Vv和Nv与对照组比较均有极显著性增大(P＜0.01)。 上述大脑皮质各部位凋亡细胞Vv和Nv变化情况:在3组缺血15min再灌注中,凋亡细胞Vv和Nv在12h显著上升(P＜0.05),海马CA1、CA2区以及顶叶皮质区在24h极显著上升(P＜0.01)。在3组缺血2h再灌注中,凋亡细胞Vv和Nv在12h极显著上升(P＜0.01),海马CA1、CA2区以及顶叶皮质区在24h极显著上升(P＜0.01)。缺血2h再灌注后6h、12h、24h组分别与缺血15min再灌注后6h、12h、24h组比较,3个缺血2h组有显著(P＜0.05)或极显著增大(P＜0.01)。 大脑皮质不同区域的凋亡细胞Vv和Nv比较:脑缺血15min再灌注后,在海马结构中,CA1区凋亡细胞Vv和Nv最大,CA2区其次,然后是CA3,最后为CA4。CA1与CA2、CA3及CA4比较均有极显著性差异(P＜0.01),CA2与CA3及CA4比较有显著性差异(P＜0.05),CA3与CA4无显著性差异(P＞0.05)。顶叶皮质区凋亡细胞Vv和Nv均高于海马和齿状回区,并且有极显著性差异(P＜0.01)。海马凋亡细胞Vv高于齿状回,而Nv无显著性差异(P＞0.05)。脑缺血2h再灌注后,CA1区凋亡细胞Vv和Nv最大,CA2区其次,然后是CA3,最后为CA4。CA1与CA2、CA3及CA4比较均有极显著性差异(P＜0.01),CA2与CA3及CA4比较有显著性差异(P＜0.05),CA3与CA4无显著性差异(P＞0.05)。顶叶皮质区凋亡细胞Vv和Nv高于海马和齿状回,并且有极显著性差异(P＜0.01)。海马凋亡细胞Vv高于齿状回,而Nv无显著性差异(P＞0.05)。 Bax免疫组化结果:Bax免疫组化阳性细胞胞浆、胞核均着色,呈棕黄色,且Bax免疫组化阳性细胞多为锥体细胞。各缺血再灌注组中上述大脑皮质各部位Bax免疫组化阳性细胞的Vv和Nv与对照组比较均有极显著性增大(P＜0.01)。 各缺血再灌注组Bax免疫组化阳性细胞Vv和Nv组间比较结果:在3组缺血15min再灌注中,Vv和Nv在12h显著上升(P＜0.05),海马CA1、CA2区以及顶叶皮质区在24h极显著上升(P＜0.01)。在3组缺血2h再灌注中,Vv和Nv在12h极显著上升(P＜0.01),海马CA1、CA2区以及顶叶皮质区在24h极显著上升(P＜0.01)。缺血2h再灌注后6h、12h、24h组分别与缺血15min再灌注后6h、12h、24h组比较,3个缺血2h组有显著(P＜0.05)或极显著增大(P＜0.01)。 Bax免疫组化阳性细胞Vv和Nv比较:脑缺血15min再灌注后,在海马结构中,CA1区Bax免疫组化阳性细胞Vv和Nv最大,CA2区其次,最后为CA3、CA4。CA1与CA2比较有显著性差异(P＜0.05),与CA3及CA4比较均有极显著性差异(P＜0.01),CA2与CA3及CA4比较有显著性差异(P＜0.05),CA3与CA4无显著性差异(P＞0.05)。与海马比较,顶叶皮质区Bax免疫组化阳性细胞Vv和Nv更大,且两者之间有极显著性差异(P＜0.01)。在齿状回未发现Bax表达。脑缺血2h再灌注后,在海马结构中,CA1区Bax免疫组化阳性细胞Vv和Nv最大,CA2区其次,然后是CA3,最后为CA4,CA1与CA2、CA3及CA4比较均有极显著性差异(P＜0.01),CA2与CA3及CA4比较有显著性差异(P＜0.05),CA3与CA4无显著性差异(P＞0.05)。顶叶皮质区Bax免疫组化阳性细胞Vv和Nv均高于海马区,且两者之间有极显著性差异(P＜0.01)。在齿状回未发现Bax表达。 各缺血再灌注组中上述大脑皮质各区域Hoechst33258荧光染色显示的凋亡细胞Vv和Nv与Bax免疫组化阳性细胞Vv和Nv比较均有显著性差异(P＜0.05)或极显著性差异(P＜0.01)。 细胞凋亡参与了脑缺血再灌注损伤,脑缺血时间和再灌注时间的增长会加重细胞凋亡的程度。大鼠脑缺血再灌注后大脑皮质不同区域细胞凋亡程度不同,在海马结构中CA1＞CA2＞CA3＼CA4,顶叶皮质区＞海马＞齿状回,而大脑左右两侧比较无明显差异。
[Abstract]:In order to observe the changes of apoptosis in different regions of cerebral cortex of rats after cerebral ischemia and reperfusion, 42 SD rats were randomly divided into control group, ischemia 15min reperfusion 6h, 12h, 24h, 2h reperfusion of 6h, 12h, 24h, and 6 in each group. The left carotid artery was ligated and the right side of the artery was clipped to the right. The model of cerebral ischemia reperfusion was constructed by the method of restoring the blood flow after the 15min and 2h of the common carotid artery. The apoptosis of hippocampal CA1, CA2, CA3, CA4 area, dentate gyrus and parietal cortex were observed by Hoechst33258 fluorescence staining and Bax immunohistochemical method, and the quantitative analysis of the volume density (Vv) and number density (Nv) was used to analyze this method. The apoptosis of cells in different regions during different periods of survival after cerebral ischemia-reperfusion.
Hoechst33258 fluorescence staining: the nucleus of apoptotic cells showed bright bright blue fluorescence under ultraviolet light. The Vv and Nv of apoptotic cells in each part of the cerebral cortex of each ischemic reperfusion group were significantly increased (P < 0.01) compared with the control group.
The changes of Vv and Nv in the apoptotic cells of all parts of the cerebral cortex: in the 3 groups of ischemic 15min reperfusion, the apoptotic cells Vv and Nv were significantly increased in 12h (P < 0.05), the hippocampus CA1, CA2 region and the parietal cortex were significantly increased in 24h (P < 0.01). The region and parietal cortex were significantly increased in 24h (P < 0.01). After ischemia 2H reperfusion, group 6h, 12h and 24h were compared with 6h, 12h and 24h groups after ischemia-reperfusion, respectively, in the 3 ischemic 2H groups (P < 0.05) or significantly increased (0.01).
Vv and Nv of apoptotic cells in different regions of the cerebral cortex: after 15min reperfusion in cerebral ischemia, the apoptotic cells in the CA1 region were the largest in the hippocampal structure, and then in the CA2 region, followed by CA3, and at last there were significant differences between CA4.CA1 and CA2, CA3 and CA4 were significantly different (0.05), and there was no significant difference between 15min and CA2. The sex difference (P > 0.05). The Vv and Nv of the apoptotic cells in the parietal cortex were higher than those in the hippocampus and dentate gyrus (P < 0.01). The Vv of the apoptotic cells in the hippocampus was higher than the dentate gyrus, but there was no significant difference between the Nv and the dentate gyrus (P > 0.05). The apoptotic cells in the CA1 region were the largest in the CA1 region, followed by the CA2 region. There were significant differences in A3 and CA4 (P < 0.01). There was a significant difference between CA2 and CA3 and CA4 (P < 0.05). There was no significant difference between CA3 and CA4 (P > 0.05). The apoptotic cells in the parietal cortex were higher than the hippocampus and dentate gyrus, and the apoptotic cells of the hippocampus were higher than the dentate gyrus, but there was no significant difference (0) .05).
Bax immunohistochemical results: the cytoplasm of Bax immunohistochemical positive cells was stained and brown yellow, and the Bax immunoreactive cells were mostly pyramidal cells. The Vv and Nv of the Bax immunoreactive cells in each part of the cerebral cortex of each ischemic reperfusion group were significantly increased (P < 0.01).
The results of comparison between Vv and Nv groups of Bax immuno positive cells in the ischemic reperfusion group: in the 3 groups of ischemic 15min reperfusion, Vv and Nv were significantly increased in 12h (P < 0.05), the hippocampus CA1, CA2 region and the parietal cortex were significantly increased in 24h (P < 0.01). The cortex of the parietal lobe increased significantly in 24h (P < 0.01). After ischemia-reperfusion, the 6h, 12h, 24h groups were compared with those of 6h, 12h and 24h groups after ischemia-reperfusion, respectively, in the 3 ischemic 2H groups (P < 0.05) or significantly increased (0.01).
Bax immunohistochemistry positive cells Vv and Nv: after cerebral ischemia 15min reperfusion, in the hippocampus structure, the CA1 region Bax immunoreactive cells Vv and Nv are the largest, CA2 region next, CA3, CA4.CA1 and CA2, there are significant differences (< 0.05). The difference (P < 0.05), CA3 and CA4 had no significant difference (P > 0.05). Compared with the hippocampus, Bax immunoreactive cells in the parietal cortex were significantly higher in Vv and Nv (P < 0.01). No Bax expression was found in the dentate gyrus. After cerebral ischemia, 2h reperfusion in the hippocampus, the CA1 region Bax immunoreactive cells and the largest Secondly, it was CA3, and finally CA4, CA1 and CA2, CA3 and CA4 were significantly different (P < 0.01), CA2 and CA3 and CA4 were significantly different (P < 0.05). There was no significant difference between CA3 and CA4 (0.05). Bax expression was not found in the dentate gyrus.
There were significant differences (P < 0.05) or very significant difference (P < 0.05) or significant difference (P < 0.01) between the apoptotic cells Vv and Nv displayed by Hoechst33258 fluorescent staining in each cerebral cortex of each ischemic reperfusion group compared with Vv and Nv of Bax immunoreactive cells.
Apoptosis is involved in cerebral ischemia reperfusion injury. The increase of cerebral ischemia time and reperfusion time will aggravate the degree of apoptosis. The degree of apoptosis in different regions of cerebral cortex in rats after cerebral ischemia-reperfusion is different. In the hippocampus structure, CA1 > CA2 > CA3 / CA4, parietal cortex area > hippocampus > dentate gyrus, but the left and right sides of the brain are compared. There is no obvious difference.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R361.2

【参考文献】

相关期刊论文 前10条

1 魏东,高春芳,戴丽;FPC恶变过程中电镜下凋亡细胞的形态学变化[J];实用医药杂志;2002年04期

2 陈红兵,童萼塘,孙圣刚,郭云良;全身亚低温对脑缺血再灌流损伤神经细胞凋亡及Caspase-3的影响[J];中国全科医学;2004年02期

3 罗焕敏;海马结构──从形态、功能到可塑性、衰老性变化[J];神经解剖学杂志;1996年02期

4 王文恭,童坦君;细胞凋亡过程中bcl-2基因的甲基化[J];中国生物化学与分子生物学报;1998年03期

5 王文恭,童坦君;小鼠成纤维细胞凋亡过程中P53与bcl-2表达的时序性[J];中国生物化学与分子生物学报;1998年03期

6 袁永辉,吴人亮,王曦,陈多恩;原位细胞凋亡TUNEL法的改进及其应用[J];陕西医学杂志;2004年07期

7 鲁利群;范建义;赵聪敏;;环境刺激对缺氧缺血性脑损伤新生大鼠学习记忆及海马病理学的影响[J];实用儿科临床杂志;2007年02期

8 李金宝;王晓琳;陈辉;熊源长;邓小明;;兴奋性氨基酸受体拮抗剂对清醒大鼠脑缺血再灌注损伤的保护作用[J];实用医学杂志;2007年07期

9 陈宝贵;赵振发;卞景芝;李洪艳;陈慧娲;赵廷浩;;回神颗粒对实验性大鼠局灶性脑缺血缺血区神经细胞凋亡的影响[J];天津中医药;2006年03期

10 李红伟,王守彪;脑缺血后细胞凋亡与PARP的关系[J];泰山医学院学报;2004年02期

，

本文编号：2054543