本文選題：線栓法　切入點(diǎn)：腦缺血再灌注損傷　出處：《蘇州大學(xué)》2014年碩士論文

【摘要】：目的：使用改良Zea Longa線栓法建立SD (Sprague-Dawley)局灶性腦缺血再灌注損傷大鼠模型，應(yīng)用人工合成E-選擇素對(duì)大鼠進(jìn)行干預(yù)，研究其對(duì)大鼠腦缺血再灌注損傷后的血腦屏障通透性、腦梗死及腦水腫體積的影響。 方法：將90只雄性SD大鼠隨機(jī)分為假手術(shù)組（A組）、模型組（B組）和人工合成E-選擇素干預(yù)組（C組），每組30只。采用改良Zea Longa線栓法建立SD大鼠腦缺血再灌注損傷模型。E-選擇素干預(yù)組在建立模型10min前從股靜脈注射E-選擇素（10mg/kg）。所有大鼠在取腦前2h從股靜脈注射2.5%伊文氏藍(lán)（0.2mg/kg）。在缺血2h再灌注3h、6h、24h、48h、72h5個(gè)時(shí)間點(diǎn)進(jìn)行神經(jīng)行為學(xué)評(píng)分、MRI掃描測(cè)定腦梗死及腦水腫的體積比、多功能酶標(biāo)儀測(cè)定腦組織中伊文氏藍(lán)含量等方法了解人工合成E-選擇素對(duì)腦缺血再灌注損傷大鼠血腦屏障通透性的干預(yù)作用。 結(jié)果：（1） A組未出現(xiàn)神經(jīng)功能缺失，B組和C組均有不同程度的神經(jīng)功能缺失，B組與C組之間部分時(shí)間點(diǎn)（3h、6h、24h）神經(jīng)功能評(píng)分存在統(tǒng)計(jì)學(xué)差異（P0.05）；（2）A組無(wú)腦水腫及梗死， B、C兩組均有不同程度腦水腫及梗死且C組各時(shí)間點(diǎn)MRI測(cè)得腦組織梗死及水腫的體積較B組小，有統(tǒng)計(jì)學(xué)意義（P0.05）；（3）A組腦組織伊文氏藍(lán)含量各時(shí)間點(diǎn)無(wú)明顯變化，B組和C組兩組變化趨勢(shì)相近，再灌注3h EB含量開(kāi)始升高，6h繼續(xù)升高，24h達(dá)到峰值，48h有所下降，，72h仍高于A組。C組各時(shí)間點(diǎn)伊文氏藍(lán)含量值均低于B組，兩組差異顯著（P0.05）。 結(jié)論：1、大鼠局灶性腦缺血再灌注損傷可導(dǎo)致血腦屏障的通透性增加；2、人工合成E-選擇素能夠減少大鼠局灶性腦缺血再灌注損傷導(dǎo)致的腦梗死及腦水腫的體積，減輕血腦屏障破壞導(dǎo)致的通透性增加，具有血腦屏障保護(hù)作用。 目的：探討人工合成E-選擇素對(duì)大鼠局部腦缺血再灌注損傷腦組織內(nèi)皮細(xì)胞間緊密連接蛋白咬合蛋白（occludin）及閉鎖小帶蛋白-1（ZO-1）含量表達(dá)及位置結(jié)構(gòu)的影響，從而初步探討人工合成E-選擇素對(duì)血腦屏障保護(hù)的作用機(jī)制。 方法：采用改良的Zea Longa線栓法建立SD大鼠局灶性腦缺血再灌注損傷模型，將90只雄性SD大鼠隨機(jī)分為假手術(shù)組（A組）、模型組（B組）和人工合成E-選擇素干預(yù)組（C組），每組30只。各組大鼠分別在腦缺血2h再灌注3h、6h、24h、48h、72h5個(gè)時(shí)間點(diǎn)提取腦組織標(biāo)本。采用免疫組化法觀察缺血區(qū)腦組織occludin及ZO-1的位置結(jié)構(gòu)及含量，Western blot法半定量測(cè)定occludin及ZO-1的含量。 結(jié)果：免疫組化：（1）A組occludin及ZO-1陽(yáng)性細(xì)胞均沿血管豐富表達(dá)。（2）B組occludin及ZO-1都于缺血再灌注3h陽(yáng)性表達(dá)數(shù)減少，24h陽(yáng)性表達(dá)數(shù)降至最低，72h陽(yáng)性表達(dá)有所增加，但仍低于A組。occludin表達(dá)的連續(xù)性在24h達(dá)到最差。（3） C組occludin及ZO-1陽(yáng)性表達(dá)數(shù)在各時(shí)間點(diǎn)均高于B組但低于A組。Occludin表達(dá)的連續(xù)性在24h也好于B組，差于A組。 Western blot半定量分析結(jié)果：（1）A組occludin及ZO-1蛋白表達(dá)水平無(wú)明顯變化。（2）B組occludin及ZO-1蛋白的變化趨勢(shì)相似，3h蛋白表達(dá)水平開(kāi)始減少，6h繼續(xù)減少，24h達(dá)到最低值，48h有所回升，72h仍低于A組。（3）C組各時(shí)間點(diǎn)occludin及ZO-1變化趨勢(shì)同B組，但蛋白表達(dá)水平均高于B組，差異均有統(tǒng)計(jì)學(xué)意義（P0.05）。 結(jié)論：1．局灶性腦缺血再灌注損傷后大鼠腦組織內(nèi)皮細(xì)胞緊密連接蛋白o(hù)ccludin及ZO-1表達(dá)減少；2．人工合成E-選擇素血腦屏障的保護(hù)作用可能與其抑制occludin及ZO-1表達(dá)的減少以及維持occludin位置和結(jié)構(gòu)有關(guān)。
[Abstract]:Objective: to establish a method using modified Zea SD plug Longa line (Sprague-Dawley) rat model of focal cerebral ischemia reperfusion injury, the application of artificial synthetic E- selectin intervention in rats, study on rat cerebral ischemia reperfusion blood brain barrier permeability after injury and effect of cerebral infarction and cerebral edema volume.
Methods: 90 male SD rats were randomly divided into sham operation group (A group), model group (group B) and synthetic E- selectin in the intervention group (C group), 30 rats in each group. The establishment of cerebral ischemia reperfusion damage in SD rat model of.E- selectin in intervention group from femoral vein injection of E- in model 10min the former using modified Zea Longa suture method (10mg/kg). All rats in the brain before 2H was injected into the femoral vein, 2.5% Evans blue (0.2mg/kg). After 2h of ischemia reperfusion 3h, 6h, 24h, 48h, 72h5 time point neurobehavioral score, cerebral infarction and cerebral determination edema MRI scan volume ratio method of Evans blue content of multifunctional microplate in brain tissue were determined to investigate the intervention effect of synthetic E- selectin on cerebral ischemia reperfusion injury of blood brain barrier permeability in rats.
Results: (1) A group without loss of nerve function, neurological deficits in different degrees of B group and C group, B group and C group between the time point (3H, 6h, 24h) there was a significant difference between the scores of neural function (P0.05); (2) A group without cerebral edema and infarction, B C, the two groups have different levels of cerebral edema and infarction and C group at each time point MRI measured the brain infarct and edema volume was smaller than that in group B, with statistical significance (P0.05); (3) the brain tissue of the A group of Evans blue content at each time point has no obvious change, B group and C group of two change similar trends, reperfusion 3H EB content began to increase, 6h continues to rise, 24h peak, 48h decreased, 72h is still higher than that of A group.C group at different time points of Evans blue content were lower than B group, there was significant difference between two groups (P0.05).
Conclusion: 1, can lead to increased permeability of the blood-brain barrier in rats with focal cerebral ischemia reperfusion injury; 2, synthetic E- selectin can reduce rat focal cerebral ischemia reperfusion injury caused by cerebral infarction and cerebral edema volume, reduce the destruction of the blood-brain barrier permeability leading to increased with protection the role of the blood-brain barrier.
Objective: To investigate the effect of synthetic E- selectin on local cerebral ischemia reperfusion injury in rat brain endothelial cell tight junction protein occludin (occludin) and -1 (ZO-1) zonula occludens protein content expression and location of structure, so as to investigate the synthetic E- mechanism of selectin on protection of blood brain barrier.
Methods: SD rats with focal cerebral ischemia reperfusion injury model using Zea Longa modified suture method, 90 male SD rats were randomly divided into sham operation group (A group), model group (group B) and synthetic E- selectin in the intervention group (C group), 30 rats in each group. All the rats were 2H in cerebral ischemia reperfusion 3h, 6h, 24h, 48h, 72h5 time points to extract brain tissue specimens. The position of structure and content by immunohistochemical staining of occludin and ZO-1 in brain tissue was observed in ischemic area, semi quantitative determination of occludin content and ZO-1 Western blot method.
Results: the results of immunohistochemistry: (1) A group occludin and ZO-1 positive cells were abundant along vascular expression. (2) B and ZO-1 in group occludin ischemia reperfusion 3H positive expression decreased, 24h expression decreased to the lowest number, the positive expression of 72h increased, but still lower than the continuous expression of A group.Occludin lowest in 24h. (3) C group occludin and ZO-1 positive expression at each time point were higher than that of B group but lower than the continuous expression of A.Occludin in 24h group or in B group, the difference in the A group.
Western blot semi quantitative analysis results: (1) A group occludin and the expression level of ZO-1 protein had no obvious change. (2) the change trends of the B group of occludin and ZO-1 protein, 3h protein expression level began to decrease, 6h 24h continued to decline, reaching the lowest value of 48h increased, 72h is still lower than that of A group (3. C) group at different time points occludin and ZO-1 changes with the B group, but the expression level was higher than that of B group, the differences were statistically significant (P0.05).
Conclusion: 1. focal cerebral ischemia reperfusion injury after rat brain endothelial cell tight junction protein occludin and ZO-1 expression decreased; protective effect of 2. synthetic E- selectin blood brain barrier may be reduced due to the inhibition of occludin and expression of ZO-1 and occludin to maintain position and structure.

【學(xué)位授予單位】：蘇州大學(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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