發(fā)布時(shí)間：2018-07-05 18:10

  本文選題：缺血預(yù)適應(yīng) + 腦缺血再灌注損傷��； 參考：《河北聯(lián)合大學(xué)》2014年碩士論文

【摘要】：目的通過觀察缺血預(yù)適應(yīng)對(duì)局灶性腦缺血再灌注后大鼠神經(jīng)功能缺損評(píng)分、腦梗死體積、病理組織學(xué)損傷的影響及缺血灶周圍皮質(zhì)區(qū)及海馬CA1區(qū)細(xì)胞色素C（CytC）、凋亡誘導(dǎo)因子（AIF）表達(dá)的變化，探討缺血預(yù)適應(yīng)的腦保護(hù)作用 方法采用線栓法制備雄性SD大鼠局灶性腦缺血再灌注改良模型，將100只大鼠隨機(jī)分為假手術(shù)組（SO組）、缺血組（MCAO組）、預(yù)適應(yīng)組（BIP組），其中MCAO組與BIP組分別于缺血2h再灌注2h、6h、12h、24h、48h、72h時(shí)間點(diǎn)處死，每時(shí)間點(diǎn)5只。每組于再灌注6h、24h、72h時(shí)間點(diǎn)各多5只大鼠做TTC染色，進(jìn)行腦梗死體積的測(cè)定；SO組于24h時(shí)間點(diǎn)處死，取5只大鼠進(jìn)行TTC染色。處死大鼠前各組均行神經(jīng)功能缺損評(píng)分，各組大鼠腦組織切片行HE染色，并觀察組織病理學(xué)改變、原位末端標(biāo)記（TUNEL染色法）觀察缺血灶周圍皮質(zhì)區(qū)及海馬CA1區(qū)凋亡神經(jīng)元數(shù)、免疫組化染色觀察缺血灶周圍皮質(zhì)區(qū)及海馬CA1區(qū)CytC、AIF陽(yáng)性神經(jīng)元的表達(dá) 結(jié)果1神經(jīng)功能缺損評(píng)分、腦梗死體積及腦組織病理形態(tài)學(xué)觀察：SO組未見神經(jīng)功能缺損癥狀、缺血灶及腦組織病理性改變。與MCAO組對(duì)比，BIP組在再灌注各時(shí)間點(diǎn)神經(jīng)功能缺損評(píng)分低、缺血灶體積百分比小、腦組織病理學(xué)改變輕。2SO組皮質(zhì)區(qū)、海馬CA1區(qū)可見1~2個(gè)TUNEL染色陽(yáng)性神經(jīng)元表達(dá)。MCAO組和BIP組缺血灶周圍皮質(zhì)區(qū)及海馬CA1區(qū)TUNEL染色陽(yáng)性神經(jīng)元數(shù)均于缺血再灌注2h開始增加，之后隨再灌注時(shí)間的延長(zhǎng)TUNEL染色陽(yáng)性神經(jīng)元數(shù)逐漸增高，缺血灶周圍皮質(zhì)區(qū)及海馬CA1區(qū)再灌注24h在各觀察時(shí)間點(diǎn)中表達(dá)水平最高，隨后隨再灌注時(shí)間點(diǎn)變化，表達(dá)有下降趨勢(shì)，組內(nèi)相鄰時(shí)間點(diǎn)比較，差異有統(tǒng)計(jì)學(xué)意義（P0.05）；在相同時(shí)間點(diǎn)BIP組缺血灶周圍皮質(zhì)區(qū)及海馬CA1區(qū)TUNEL染色陽(yáng)性神經(jīng)元數(shù)量較MCAO組明顯減少，差異有統(tǒng)計(jì)學(xué)意義（P0.05）；與SO組比較，，MCAO組和BIP組各時(shí)間點(diǎn)TUNEL染色陽(yáng)性神經(jīng)元表達(dá)均增高，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。3SO組皮質(zhì)區(qū)、海馬CA1區(qū)均可見少量CytC陽(yáng)性神經(jīng)元表達(dá)；MCAO組和BIP組缺血灶周圍皮質(zhì)區(qū)及海馬CA1區(qū)CytC陽(yáng)性神經(jīng)元表達(dá)均于缺血再灌注2h開始增加，隨再灌注時(shí)間的變化CytC陽(yáng)性神經(jīng)元表達(dá)呈增高趨勢(shì)，再灌注24h在各時(shí)間點(diǎn)表達(dá)水平最高，組內(nèi)相鄰時(shí)間點(diǎn)比較，差異有統(tǒng)計(jì)學(xué)意義（P0.05）；在相同時(shí)間點(diǎn)BIP組缺血灶周圍皮質(zhì)區(qū)及海馬CA1區(qū)CytC陽(yáng)性神經(jīng)元表達(dá)數(shù)量較MCAO組明顯減少，差異有統(tǒng)計(jì)學(xué)意義（P0.05）；與SO組比較，MCAO組和BIP組各時(shí)間點(diǎn)CytC陽(yáng)性神經(jīng)元表達(dá)均增高，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。4SO組皮質(zhì)區(qū)、海馬CA1區(qū)均有少量AIF陽(yáng)性神經(jīng)元表達(dá)；MCAO組和BIP組缺血灶周圍皮質(zhì)區(qū)及海馬CA1區(qū)AIF陽(yáng)性神經(jīng)元表達(dá)趨勢(shì)基本一致，均于缺血再灌注2h表達(dá)開始增加，之后隨時(shí)間的延長(zhǎng)表達(dá)逐漸增多，在皮質(zhì)區(qū)再灌注48h在各時(shí)間點(diǎn)表達(dá)水平最高，海馬CA1區(qū)于再灌注24h在各時(shí)間點(diǎn)表達(dá)水平最高，組內(nèi)相鄰時(shí)間點(diǎn)比較，差異有統(tǒng)計(jì)學(xué)意義（P0.05）；在相同時(shí)間點(diǎn)BIP組缺血灶周圍皮質(zhì)區(qū)及海馬CA1區(qū)AIF陽(yáng)性神經(jīng)元表達(dá)數(shù)量較MCAO組明顯減少，差異有統(tǒng)計(jì)學(xué)意義（P0.05）；與SO組比較，MCAO組和BIP組各時(shí)間點(diǎn)AIF陽(yáng)性神經(jīng)元表達(dá)均增高，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。 結(jié)論1缺血預(yù)適應(yīng)后局灶性腦缺血再灌注大鼠神經(jīng)功能缺損評(píng)分降低、腦梗死體積減小、腦組織病理組織學(xué)損傷減輕，有效地減輕了腦缺血再灌注損傷。2缺血預(yù)適應(yīng)后局灶性腦缺血再灌注大鼠缺血灶周圍皮質(zhì)區(qū)及海馬CA1區(qū)神經(jīng)細(xì)胞凋亡減少，起到腦保護(hù)作用。3缺血預(yù)適應(yīng)后局灶性腦缺血再灌注大鼠缺血灶周圍皮質(zhì)區(qū)及海馬CA1區(qū)CytC、AIF的表達(dá)減少，可能通過抑制CytC、AIF的表達(dá)，抑制凋亡的發(fā)生
[Abstract]:Objective To observe the effects of ischemic preconditioning on neurological deficit score, cerebral infarction volume, histopathological injury, cytochrome C (CytC) and apoptosis inducible factor (AIF) expression in the cortex and hippocampus CA1 area around the ischemic area, and to explore the protective effect of ischemic preconditioning on cerebral protection.
Methods the improved model of focal cerebral ischemia reperfusion in male SD rats was prepared by thread emboli, and 100 rats were randomly divided into sham operation group (group SO), ischemia group (group MCAO) and pre adaptation group (group BIP), of which group MCAO and BIP were reperfusion with 2H, 6h, 12h, 24h, 48h, and 5 rats at each time point. At the time point, 5 rats were stained with TTC, and the volume of cerebral infarction was measured. Group SO was killed at the time point of 24h and 5 rats were stained with TTC. The nerve function defect scores were performed in each group before the death of rats. The brain tissue sections of each group were stained with HE, and the histopathological changes were observed, and the site terminal labeling (TUNEL staining) was used to observe the focal ischemia. The number of apoptotic neurons in the peripheral cortex and hippocampus CA1 area was observed. The expression of CytC and AIF positive neurons in the cortex and hippocampus CA1 area of ischemic foci was observed by immunohistochemical staining.
Results 1 nerve function defect score, cerebral infarction volume and brain tissue pathomorphology observation: in group SO, no neurological deficit symptoms, ischemic foci and pathological changes of brain tissue were not found. Compared with group MCAO, the score of nerve function defect in group BIP was low, the percentage of ischemic focus was small, and the pathological changes of brain tissue were light in group.2SO. In the CA1 region of the hippocampus, the expression of 1~2 TUNEL staining positive neurons expressed in the.MCAO group and the number of TUNEL staining positive neurons in the cortical area around the ischemic foci and the CA1 region of the BIP group began to increase, and the number of positive neurons in the TUNEL staining increased gradually with the time of reperfusion, and the cortical area around the ischemic focus and the CA1 region of the hippocampus were reformed. The expression level of perfusion 24h was the highest in each observation time point, then the expression decreased with the time point of reperfusion, and the difference was statistically significant (P0.05). The number of TUNEL staining positive neurons in the cortex and hippocampus CA1 region of BIP group at the same time point was significantly lower than that in the MCAO group, and the difference was found. Statistical significance (P0.05); compared with group SO, the expression of TUNEL staining positive neurons in each time point of group MCAO and BIP increased, the difference was statistically significant (P0.05) in the cortical area of the.3SO group, and the expression of a small amount of CytC positive neurons in the hippocampal CA1 area; the expression of the positive neurons in the peripheral cortex and the hippocampal CA1 region of the MCAO and BIP groups were all in the MCAO group and the BIP group. The expression of CytC positive neurons was increased with the change of reperfusion time, and the expression level of 24h at each time point was the highest. The difference was statistically significant (P0.05) in the adjacent time points in the group (P0.05), and the expression of CytC positive neurons in the peripheral cortex and CA1 region of the hippocampus at the same time point was in the same time point. The expression of CytC positive neurons in the BIP group of BIP group and the hippocampal CytC positive neurons were in the same time point. Compared with group MCAO, the difference was statistically significant (P0.05). Compared with group SO, the expression of CytC positive neurons in each time point of group MCAO and BIP group increased, the difference was statistically significant (P0.05) in the cortex area of.4SO group, and a small amount of AIF positive neurons in the CA1 region of the hippocampus, and the MCAO group and the cortex of the ischemic foci in the BIP group and the hippocampal region. The expression trend of sexual neurons was basically the same, the expression of 2h in ischemic reperfusion began to increase, and then increased with time. The expression level of 48h in cortical area was the highest at all time points, and the expression level of 24h in hippocampus CA1 region was the highest at all time points, and the difference was statistically significant in the adjacent time points in the group (P0.05 In the same time point, the number of AIF positive neurons in the cortex and hippocampal CA1 area of the BIP group was significantly lower than that in the MCAO group, and the difference was statistically significant (P0.05). Compared with the SO group, the expression of AIF positive neurons in each time point of group MCAO and BIP group increased, and the difference had the significance (P0.05).
Conclusion 1 after ischemic preconditioning, the score of neural function defect in focal cerebral ischemia reperfusion rats decreased, the volume of cerebral infarction decreased, and the injury of brain histopathology decreased, which effectively alleviated the cerebral ischemia reperfusion injury of cerebral ischemia reperfusion injury in the cortex and CA1 region of the hippocampus after.2 ischemic preconditioning. The expression of CytC and AIF in the cortex and CA1 region of the hippocampus of rats with focal cerebral ischemia reperfusion after ischemic preconditioning.3 decreased, and the expression of AIF may be inhibited by inhibiting the expression of CytC and AIF and inhibiting the occurrence of apoptosis.
【學(xué)位授予單位】：河北聯(lián)合大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R743.3

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