本文選題：缺血預(yù)處理 + 缺血再灌注損傷��； 參考：《河北北方學院》2017年碩士論文

【摘要】：隨著人口老齡化的趨勢加快,腦血管病已經(jīng)成為我國第一位致殘和死亡原因,對該病的研究一直是科研和臨床研究的熱點。腦缺血預(yù)處理(cerebral ischemic preconditioning,CIP)是指對腦組織采用缺血處理,如一次或多次短暫性、非致死性腦缺血再灌注(ischemia-reperfusion,I/R)刺激,啟動腦組織產(chǎn)生內(nèi)源性保護機制,將對致死性的缺血產(chǎn)生顯著的耐受,從而減弱或阻止腦缺血缺氧引起的瀑布反應(yīng)。炎性反應(yīng)在缺血再灌注損傷的機制中占重要地位,包括一系列炎癥細胞和炎癥因子的參與,還涉及到固有免疫系統(tǒng)和適應(yīng)性免疫系統(tǒng)的激活,起到了免疫調(diào)節(jié)和趨化作用,因此減輕炎癥反應(yīng)是治療缺血性腦血管病的重要環(huán)節(jié)。炎癥過程中,高遷移率族蛋白B1(high mobility group protein,HMGB1)主動分泌至胞外,與晚期糖基化終末產(chǎn)物受體(receptor for advanced glycation end-products,RAGE)和Toll樣受體4(Toll-like receptor 4,TLR4)結(jié)合,最終激活核因子κB(Nuclear factor-kappa B,NF-κB),從而介導炎性反應(yīng)。本研究通過建立大鼠局灶性CIP模型,深入研究RAGE和TLR4在CIP的生物學作用及可能機制,為再灌注損傷的臨床防治提供新的靶點。將198只Sprague Dawley成年雄性健康大鼠隨機分為假手術(shù)組(Sham,n=66)、缺血預(yù)處理組(CIP,n=66)和缺血再灌注組(I/R,n=66),采用改良的Zea-Longa方法制備大鼠大腦中動脈栓塞(MCAO)模型,缺血再灌注組給予缺血2小時后恢復(fù)血流,缺血預(yù)處理組采用二次線栓法,缺血10分鐘為預(yù)處理措施,72小時后給予缺血2小時再恢復(fù)腦部血流,假手術(shù)組僅分離頸總動脈,不給于栓塞處理。隨機分為再灌注后0.5 d,1 d,2 d,3 d,7 d共5個時間點,于再灌注1d時每組隨機選取6只采用TTC染色法用于測定腦梗死體積,各個時間點采用Zea Longa評分標準進行神經(jīng)功能評分,通過免疫組織化學染色法測定RAGE和TLR4蛋白水平的表達變化,定量即時聚合酶鏈鎖反應(yīng)(Real-time Quantitative Polymerase Chain Reaction,RT-qPCR)法測定RAGE和TLR4 mRNA表達。本研究發(fā)現(xiàn),Sham組無梗死灶,CIP組在1 d時腦梗死體積為明顯低于I/R組(P0.05);Sham組無神經(jīng)功能缺損,CIP組大鼠在0.5 d、1 d、2 d、3 d、7 d的神經(jīng)功能行為缺陷評分與I/R組大鼠比較明顯減低,有顯著性差異(P0.05);與sham組相比,CIP組與IR組缺血半暗帶區(qū)RAGE陽性細胞數(shù)顯著升高(P0.05),CIP組較I/R組的表達明顯降低(P0.05),TLR4陽性細胞數(shù)也在缺血半暗帶區(qū)明顯升高,CIP組與IR組均顯著高于sham組,CIP組低于I/R組,差異顯著(P0.05)。CIP組和I/R組RAGE、TLR4 mRNA表達均顯著高于sham組(P0.05),均于1 d時達高峰,隨再灌注時間的延長兩者表達逐漸下降,CIP組較I/R組的表達均明顯降低(P0.05)。因此,腦缺血再灌注損傷可激活炎癥介質(zhì),產(chǎn)生級聯(lián)反應(yīng),CIP可使大腦產(chǎn)生缺血耐受,明顯減輕炎癥反應(yīng),并減小腦梗死體積,RAGE、TLR4在缺血損傷過程中表達上調(diào),腦缺血耐受機制可能與下調(diào)缺血再灌注損傷時RAGE和TLR4的表達有關(guān)。
[Abstract]:With the aging of the population, cerebrovascular disease has become the first cause of disability and death in China, the research on the disease has been a hot spot in scientific research and clinical research. Cerebral ischemic preconditioning (cerebral ischemic preconditioning) refers to the use of ischemic treatment in brain tissues, such as once or more transient, non-fatal cerebral ischemia-reperfusion (I- / R) stimulation to activate endogenous protective mechanisms in brain tissue, which will produce significant tolerance to fatal ischemia. This weakens or blocks the waterfall response caused by cerebral ischemia and hypoxia. Inflammatory response plays an important role in the mechanism of ischemia-reperfusion injury, including the involvement of a series of inflammatory cells and inflammatory factors, as well as the activation of the innate and adaptive immune system, which plays an important role in immune regulation and chemotaxis. Therefore, reducing inflammatory reaction is an important link in the treatment of ischemic cerebrovascular disease. In the process of inflammation, the high mobility group protein B1 (high mobility group protein HMGB1 is secreted into the extracellular cells, which binds to the late glycation end product receptor (receptor for advanced glycation end-products rage and Toll-like receptor 4 (TLR4), and finally activates nuclear factor-kappa BNF- 魏 B, which mediates the inflammatory response. In order to provide a new target for clinical prevention and treatment of reperfusion injury, the biological function and possible mechanism of rage and TLR4 in CIP were studied by establishing focal CIP model in rats. 198 Sprague Dawley adult male healthy rats were randomly divided into three groups: sham operation group (Shambun 66), ischemic preconditioning group (CIP) and ischemia reperfusion group (I / R). The model of middle cerebral artery embolization (MCAO) was established by modified Zea-Longa method. In the ischemia reperfusion group, the blood flow was recovered after 2 hours of ischemia, and in the ischemic preconditioning group, the cerebral blood flow was recovered after 72 hours of ischemia for 72 hours, and the common carotid artery was only separated in the sham operation group. No embolization. The rats were randomly divided into 5 time points at 0.5 d ~ 1 d ~ 2 d ~ 3 d ~ 7 d after reperfusion. 6 rats in each group were randomly selected to measure the volume of cerebral infarction by TTC staining on the 1st day after reperfusion. The neurological function was evaluated by Zea Longa scoring standard at each time point. The expression of rage and TLR4 protein was detected by immunohistochemical staining, and the expression of rage and TLR4 mRNA was detected by Real-time quantitative Polymerase chain reaction (RT-PCR). It was found that the cerebral infarct volume in the CIP group without infarction was significantly lower than that in the I / R group at 1 day (P0.05). The neurological behavioral impairment scores in the CIP group were significantly lower than those in the I / R group at 0.5 d, 1 d, 2 d, 3 d and 7 d, respectively, compared with those in the I / R group. Compared with sham group and IR group, the number of rage positive cells in ischemic penumbra increased significantly (P0.05) the expression of rage positive cells in CIP group was significantly lower than that in I / R group (P0.05), and the number of TLR4 positive cells in ischemic penumbra was also significantly increased in sham group and IR group. Higher than the sham group, lower than the I / R group, The expression of RAGEN TLR4 mRNA in CIP group and I / R group was significantly higher than that in sham group (P0.05), and reached its peak at 1 day. The expression of RAGETLR4 mRNA in CIP group decreased gradually with the prolongation of reperfusion time (P0.05), and the expression of RAGETLR4 mRNA in CIP group was significantly lower than that in I / R group (P0.05). Therefore, cerebral ischemia-reperfusion injury can activate inflammatory mediators, produce cascade reaction CIP can induce cerebral ischemic tolerance, significantly reduce inflammatory response, and reduce the volume of cerebral infarction and up regulate the expression of RAGEN TLR4 during the process of ischemic injury. The mechanism of cerebral ischemic tolerance may be related to down-regulating the expression of rage and TLR4 in ischemic reperfusion injury.
【學位授予單位】：河北北方學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R743

【參考文獻】

相關(guān)期刊論文 前8條

1 李珊珊;李國營;;缺血性腦卒中中藥防治的研究進展[J];解剖學研究;2016年03期

2 單嬌;李宏宇;劉國峰;楊玄;董偉;簡偉研;鄧芙蓉;郭新彪;;大氣污染對中老年高血壓和心腦血管疾病患者衛(wèi)生服務(wù)需求的影響:基于CHARLS數(shù)據(jù)的分析[J];北京大學學報(醫(yī)學版);2016年03期

3 王文;隋輝;陳偉偉;;我國心血管病防治的現(xiàn)狀與策略[J];醫(yī)學研究雜志;2015年09期

4 莊微;劉挺松;;晚期糖基化終末產(chǎn)物受體在心血管疾病中的研究概況[J];東南國防醫(yī)藥;2014年06期

5 王伊龍;趙性泉;王擁軍;陳盼;張倩;;腦卒中神經(jīng)保護劑的治療現(xiàn)狀和未來[J];中國新藥雜志;2011年11期

6 ;TLR4 signaling induced TLR2 expression in the process of mimic cerebral ischemia/reperfusion in vitro[J];Science China(Life Sciences);2010年02期

7 李軍;曹紅;連慶泉;王耀岐;曾因明;姚尚龍;曾邦雄;;ERK通路在腦缺血及缺血預(yù)處理沙土鼠海馬神經(jīng)元中的作用[J];中國應(yīng)用生理學雜志;2008年02期

8 郝玉曼,羅祖明,周東;局灶預(yù)缺血誘導腦缺血耐受的動物模型[J];中風與神經(jīng)疾病雜志;2003年02期

，

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