本文選題：雷帕霉素 + 腦缺血　； 參考：《濱州醫(yī)學(xué)院》2014年碩士論文

【摘要】：引文腦缺血再灌注后梗死灶周圍的腦組織存在炎癥反應(yīng)及水腫。炎癥反應(yīng)可以導(dǎo)致腦細(xì)胞受損,從而導(dǎo)致并加重腦水腫。炎癥反應(yīng)中的某些炎癥因子可以影響腦組織中AQP4及MMP9蛋白的表達(dá)。AQP4在血腦屏障的水分子轉(zhuǎn)運(yùn)中起重要作用,MMP9在BBB基底膜破壞中起重要作用。這兩種蛋白表達(dá)增多時(shí)可以加重腦缺血再灌注后腦水腫的程度,嚴(yán)重影響患者的病情變化及預(yù)后。目的研究大鼠腦缺血再灌注同時(shí)應(yīng)用雷帕霉素對(duì)大鼠腦梗死體積、腦水腫程度、BBB受損程度及病理學(xué)變化、AQP4及MMP9蛋白的表達(dá)量等的影響,探討雷帕霉素對(duì)大鼠局灶性腦缺血再灌注損傷后的腦水腫是否具有保護(hù)作用,并初步分析雷帕霉素發(fā)揮保護(hù)的機(jī)制。方法將72只成年雄性SD大鼠隨機(jī)分為三組,雷帕霉素(rapamycin, RAPA)組、二甲基亞砜(dimethyl sulfoxide, DMSO)組和假手術(shù)(Sham)組,每組24只。在激光多普勒血流儀(laser doppler flowmeter, LDF)實(shí)時(shí)監(jiān)測(cè)下,對(duì)RAPA組、DMSO組大鼠應(yīng)用線栓法制作大腦中動(dòng)脈閉塞(MCAO)模型,缺血1h后行再灌注,RAPA組、DMSO組分別于再灌注開始時(shí)經(jīng)腹膜腔注射250 μg/ml的RAPA稀釋液(250 μg/kg體重)、DMSO (1 ml/kg體重)；Sham組大鼠只模擬手術(shù),不插入栓線。各組大鼠于麻醉清醒后進(jìn)行神經(jīng)功能缺損評(píng)分,于再灌注48 h后取材行腦梗死體積(6只)、腦組織含水量(6只)、血腦屏障(blood-brain barrier, BBB)通透性的測(cè)定(6只),光鏡、電鏡對(duì)腦組織進(jìn)行形態(tài)學(xué)觀察,免疫組織化學(xué)法檢測(cè)大腦皮質(zhì)水通道蛋4 (aquaporin 4, AQP4)和金屬基質(zhì)蛋白酶9 (matrix metalloproteinase 9, MMP9)的表達(dá)(6只)。結(jié)果①RAPA組腦梗死體積小于DMSO組(P0.05)。 ②RAPA組腦組織含水量低于DMSO組(P0.05)。 ③RAP A組伊文思藍(lán)(Evans blue, EB) BBB的透出量明顯低于DMSO組(P0.01)。④MCAO模型組腦組織損傷位于左側(cè)額葉及頂葉大腦皮層。DMSO組梗死區(qū)域內(nèi)可見大量的正常神經(jīng)元變性、壞死,甚至核固縮、尼氏體消失,形成“紅色神經(jīng)元”(嗜酸性變性)；腦組織水腫明顯,大量中性粒細(xì)胞浸潤(rùn),毛細(xì)血管周圍間隙明顯增大。RAPA組梗死范圍及缺血半暗帶范圍明顯小于DMSO組,腦組織缺血及梗死等病理變化程度較DMSO組輕。假手術(shù)組腦組織結(jié)構(gòu)基本正常。⑤DMSO組梗死區(qū)血腦屏障完整性嚴(yán)重受損,毛細(xì)血管管腔閉鎖,管周水腫明顯,基膜欠完整。RAPA組BBB破壞程度較對(duì)照組輕。⑥RAPA組大腦半球額、頂葉皮層缺血半暗帶區(qū)AQP4、MMP9的表達(dá)均明顯低于DMSO組(P0.01)。結(jié)論再灌注同時(shí)應(yīng)用雷帕霉素可以減小腦梗死體積,減輕腦水腫程度,降低BBB的病理學(xué)損傷程度,還可以降低腦組織中AQP4和MMP9的表達(dá)。雷帕霉素可能是通過抑制免疫級(jí)聯(lián)反應(yīng),下調(diào)腦組織中AQP4和MMP9的表達(dá),改善腦水腫,從而對(duì)缺血再灌注損傷后的腦組織產(chǎn)生保護(hù)作用。
[Abstract]:There are inflammatory reaction and edema in the brain tissue around the infarct after cerebral ischemia reperfusion. Inflammation can cause brain cells to be damaged, leading to and exacerbating brain edema. Some inflammatory factors may affect the expression of AQP4 and MMP9 protein in brain tissue. AQP4 plays an important role in the water molecule transport of blood-brain barrier. MMP9 plays an important role in the destruction of BBB basement membrane. The increased expression of these two proteins can aggravate the degree of cerebral edema after cerebral ischemia reperfusion, and seriously affect the state of disease and prognosis of the patients. Objective to study the effects of rapamycin on cerebral infarction volume, cerebral edema and pathological changes in rats with cerebral ischemia-reperfusion and the expression of AQP4 and MMP9 protein. To investigate the protective effect of rapamycin on brain edema after focal cerebral ischemia reperfusion injury in rats, and to analyze the protective mechanism of rapamycin. Methods Seventy-two adult male Sprague-Dawley rats were randomly divided into three groups: rapamycin (rapamycin) group, dimethyl sulfoxide (DMSO) group and sham operation group (24 rats in each group). The model of middle cerebral artery occlusion (MCAO) was established by using the method of thread occlusion in rats of RAPA group under the real-time monitoring of laser doppler flowmeter, LDF) with laser Doppler flow meter. At the beginning of reperfusion, the rats in the rapa group were injected with 250 渭 g/ml RAPA diluted solution (250 渭 g/kg) at the beginning of reperfusion, respectively. The rats in the control group were given DMSO 1 ml/kg weight / body weight respectively. The rats in the sham group were treated with simulated operation and no thrombus was inserted. The rats in each group were assessed for neurological impairment after anaesthesia. After 48 hours of reperfusion, 6 rats with cerebral infarction volume, 6 rats with cerebral tissue water content and 6 rats with blood-brain barrier permeability were measured under light microscope. The expression of aquaporin 4 (AQP4) and metalloproteinase 9 matrix metalloproteinase 9 (MMP9) in cerebral cortical aquaporin 4 (AQP4) and metalloproteinase 9 (MMP9) were detected by immunohistochemistry. Results the volume of cerebral infarction in 1RAPA group was smaller than that in DMSO group (P 0.05). The brain tissue water content in 2RAPA group was lower than that in DMSO group (P 0.05). The permeability of Evans blue, EB) BBB in 3RAP A group was significantly lower than that in DMSO group (P 0.01) .4MCAO model group was located in left frontal lobe and parietal area. In lobar cortex. DMSO group, a large number of normal neuronal degeneration could be seen in the infarcted area. Necrosis, even nuclear shrinkage, the disappearance of the Nissl body, and the formation of "red neurons" (eosinophilic denaturation); brain tissue edema, massive neutrophil infiltration, The infarct size and ischemic penumbra in the rapa group were significantly smaller than those in the DMSO group, and the pathological changes of cerebral ischemia and infarction were lighter in the rapa group than in the DMSO group. In sham-operation group, the integrity of blood-brain barrier in infarcted area was seriously damaged, capillary lumen atresia, peritube edema was obvious, and the degree of BBB damage in rapa group was less than that in control group. The expression of AQP4 MMP9 in ischemic penumbra of parietal cortex was significantly lower than that in DMSO group. Conclusion reperfusion with rapamycin can reduce the volume of cerebral infarction, reduce the degree of cerebral edema, reduce the degree of pathological injury of BBB, and decrease the expression of AQP4 and MMP9 in brain tissue. Rapamycin may decrease the expression of AQP4 and MMP9 in brain tissue by inhibiting immune cascade reaction, and improve brain edema, which may have protective effect on brain tissue after ischemia-reperfusion injury.
【學(xué)位授予單位】：濱州醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R743;R742

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相關(guān)期刊論文 前2條

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