本文選題：腦缺血再灌注 + 凋亡��； 參考：《南昌大學》2008年碩士論文

【摘要】： 為了觀察腦缺血再灌注后不同存活時間內(nèi)大鼠大腦皮質(zhì)不同區(qū)域細胞凋亡的變化情況,本實驗選取42只SD大鼠,將其隨機分為對照組,缺血15min再灌注6h、12h、24h組,缺血2h再灌注6h、12h、24h組,每組6只。通過結(jié)扎左側(cè)頸總動脈,動脈夾夾閉右側(cè)頸總動脈15min和2h后松開動脈夾恢復(fù)血流的方法構(gòu)建腦缺血再灌注模型。采用Hoechst33258熒光染色和Bax免疫組化方法觀察海馬CA1、CA2、CA3、CA4區(qū)、齒狀回及頂葉皮質(zhì)區(qū)細胞凋亡情況,用體視學參數(shù)——體積密度(Vv)和數(shù)密度(Nv)定量地分析這些區(qū)域細胞在腦缺血再灌注后不同存活時間內(nèi)的凋亡情況。結(jié)果如下: Hoechst33258熒光染色:凋亡細胞細胞核在紫外光下呈現(xiàn)出明亮鮮艷的藍色熒光。各缺血再灌注組中上述大腦皮質(zhì)各部位凋亡細胞Vv和Nv與對照組比較均有極顯著性增大(P＜0.01)。 上述大腦皮質(zhì)各部位凋亡細胞Vv和Nv變化情況:在3組缺血15min再灌注中,凋亡細胞Vv和Nv在12h顯著上升(P＜0.05),海馬CA1、CA2區(qū)以及頂葉皮質(zhì)區(qū)在24h極顯著上升(P＜0.01)。在3組缺血2h再灌注中,凋亡細胞Vv和Nv在12h極顯著上升(P＜0.01),海馬CA1、CA2區(qū)以及頂葉皮質(zhì)區(qū)在24h極顯著上升(P＜0.01)。缺血2h再灌注后6h、12h、24h組分別與缺血15min再灌注后6h、12h、24h組比較,3個缺血2h組有顯著(P＜0.05)或極顯著增大(P＜0.01)。 大腦皮質(zhì)不同區(qū)域的凋亡細胞Vv和Nv比較:腦缺血15min再灌注后,在海馬結(jié)構(gòu)中,CA1區(qū)凋亡細胞Vv和Nv最大,CA2區(qū)其次,然后是CA3,最后為CA4。CA1與CA2、CA3及CA4比較均有極顯著性差異(P＜0.01),CA2與CA3及CA4比較有顯著性差異(P＜0.05),CA3與CA4無顯著性差異(P＞0.05)。頂葉皮質(zhì)區(qū)凋亡細胞Vv和Nv均高于海馬和齒狀回區(qū),并且有極顯著性差異(P＜0.01)。海馬凋亡細胞Vv高于齒狀回,而Nv無顯著性差異(P＞0.05)。腦缺血2h再灌注后,CA1區(qū)凋亡細胞Vv和Nv最大,CA2區(qū)其次,然后是CA3,最后為CA4。CA1與CA2、CA3及CA4比較均有極顯著性差異(P＜0.01),CA2與CA3及CA4比較有顯著性差異(P＜0.05),CA3與CA4無顯著性差異(P＞0.05)。頂葉皮質(zhì)區(qū)凋亡細胞Vv和Nv高于海馬和齒狀回,并且有極顯著性差異(P＜0.01)。海馬凋亡細胞Vv高于齒狀回,而Nv無顯著性差異(P＞0.05)。 Bax免疫組化結(jié)果:Bax免疫組化陽性細胞胞漿、胞核均著色,呈棕黃色,且Bax免疫組化陽性細胞多為錐體細胞。各缺血再灌注組中上述大腦皮質(zhì)各部位Bax免疫組化陽性細胞的Vv和Nv與對照組比較均有極顯著性增大(P＜0.01)。 各缺血再灌注組Bax免疫組化陽性細胞Vv和Nv組間比較結(jié)果:在3組缺血15min再灌注中,Vv和Nv在12h顯著上升(P＜0.05),海馬CA1、CA2區(qū)以及頂葉皮質(zhì)區(qū)在24h極顯著上升(P＜0.01)。在3組缺血2h再灌注中,Vv和Nv在12h極顯著上升(P＜0.01),海馬CA1、CA2區(qū)以及頂葉皮質(zhì)區(qū)在24h極顯著上升(P＜0.01)。缺血2h再灌注后6h、12h、24h組分別與缺血15min再灌注后6h、12h、24h組比較,3個缺血2h組有顯著(P＜0.05)或極顯著增大(P＜0.01)。 Bax免疫組化陽性細胞Vv和Nv比較:腦缺血15min再灌注后,在海馬結(jié)構(gòu)中,CA1區(qū)Bax免疫組化陽性細胞Vv和Nv最大,CA2區(qū)其次,最后為CA3、CA4。CA1與CA2比較有顯著性差異(P＜0.05),與CA3及CA4比較均有極顯著性差異(P＜0.01),CA2與CA3及CA4比較有顯著性差異(P＜0.05),CA3與CA4無顯著性差異(P＞0.05)。與海馬比較,頂葉皮質(zhì)區(qū)Bax免疫組化陽性細胞Vv和Nv更大,且兩者之間有極顯著性差異(P＜0.01)。在齒狀回未發(fā)現(xiàn)Bax表達。腦缺血2h再灌注后,在海馬結(jié)構(gòu)中,CA1區(qū)Bax免疫組化陽性細胞Vv和Nv最大,CA2區(qū)其次,然后是CA3,最后為CA4,CA1與CA2、CA3及CA4比較均有極顯著性差異(P＜0.01),CA2與CA3及CA4比較有顯著性差異(P＜0.05),CA3與CA4無顯著性差異(P＞0.05)。頂葉皮質(zhì)區(qū)Bax免疫組化陽性細胞Vv和Nv均高于海馬區(qū),且兩者之間有極顯著性差異(P＜0.01)。在齒狀回未發(fā)現(xiàn)Bax表達。 各缺血再灌注組中上述大腦皮質(zhì)各區(qū)域Hoechst33258熒光染色顯示的凋亡細胞Vv和Nv與Bax免疫組化陽性細胞Vv和Nv比較均有顯著性差異(P＜0.05)或極顯著性差異(P＜0.01)。 細胞凋亡參與了腦缺血再灌注損傷,腦缺血時間和再灌注時間的增長會加重細胞凋亡的程度。大鼠腦缺血再灌注后大腦皮質(zhì)不同區(qū)域細胞凋亡程度不同,在海馬結(jié)構(gòu)中CA1＞CA2＞CA3＼CA4,頂葉皮質(zhì)區(qū)＞海馬＞齒狀回,而大腦左右兩側(cè)比較無明顯差異。
[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.
【學位授予單位】：南昌大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R361.2

【參考文獻】

相關(guān)期刊論文 前10條

1 魏東,高春芳,戴麗;FPC惡變過程中電鏡下凋亡細胞的形態(tài)學變化[J];實用醫(yī)藥雜志;2002年04期

2 陳紅兵,童萼塘,孫圣剛,郭云良;全身亞低溫對腦缺血再灌流損傷神經(jīng)細胞凋亡及Caspase-3的影響[J];中國全科醫(yī)學;2004年02期

3 羅煥敏;海馬結(jié)構(gòu)──從形態(tài)、功能到可塑性、衰老性變化[J];神經(jīng)解剖學雜志;1996年02期

4 王文恭,童坦君;細胞凋亡過程中bcl-2基因的甲基化[J];中國生物化學與分子生物學報;1998年03期

5 王文恭,童坦君;小鼠成纖維細胞凋亡過程中P53與bcl-2表達的時序性[J];中國生物化學與分子生物學報;1998年03期

6 袁永輝,吳人亮,王曦,陳多恩;原位細胞凋亡TUNEL法的改進及其應(yīng)用[J];陜西醫(yī)學雜志;2004年07期

7 魯利群;范建義;趙聰敏;;環(huán)境刺激對缺氧缺血性腦損傷新生大鼠學習記憶及海馬病理學的影響[J];實用兒科臨床雜志;2007年02期

8 李金寶;王曉琳;陳輝;熊源長;鄧小明;;興奮性氨基酸受體拮抗劑對清醒大鼠腦缺血再灌注損傷的保護作用[J];實用醫(yī)學雜志;2007年07期

9 陳寶貴;趙振發(fā);卞景芝;李洪艷;陳慧媧;趙廷浩;;回神顆粒對實驗性大鼠局灶性腦缺血缺血區(qū)神經(jīng)細胞凋亡的影響[J];天津中醫(yī)藥;2006年03期

10 李紅偉,王守彪;腦缺血后細胞凋亡與PARP的關(guān)系[J];泰山醫(yī)學院學報;2004年02期

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