【摘要】：目的 腎上腺髓質(zhì)素(ADM)是由52個(gè)氨基酸組成的血管活性多肽,屬于降鈣素基因相關(guān)肽(CGRP)家族,與多種組織的缺血再灌注損傷有關(guān)。有學(xué)者證實(shí)在中樞神經(jīng)系統(tǒng)有ADM的表達(dá),并與腦缺血再灌注損傷有一定關(guān)系。也有持相反意見的研究報(bào)道。所以我們選用大鼠局灶性腦缺血再灌注模型,應(yīng)用免疫組織化學(xué)方法、原位雜交組織化學(xué)法、逆轉(zhuǎn)錄—聚合酶鏈?zhǔn)?RT-PCR)分析方法等,驗(yàn)證正常大鼠腦組織是否有ADM及ADM mRNA的表達(dá)及具體表達(dá)的部位;觀察ADM及ADM mRNA在局灶性缺血再灌注大鼠腦組織中的表達(dá)變化規(guī)律;探討ADM對(duì)局灶性缺血再灌注大鼠神經(jīng)元凋亡、梗死體積及Egr-1 mRNA的影響,進(jìn)一步研究ADM在局灶性缺血再灌注腦損傷中的作用。 方法 健康雄性SD大鼠186只,體重200-250g,由中國(guó)醫(yī)科大學(xué)實(shí)驗(yàn)動(dòng)物中心提供。隨機(jī)分為正常組(n=12),假手術(shù)組(n=12),局灶性腦缺血2h再灌注2h組(n=18)、4h組(n=18)、22h組(n=54)、46h組(n=18)、70h組(n=18)、118h組(n=18)、166h組(n=18)。其中再灌注22h組又分為:股靜脈注射ADM組(n=12),頸內(nèi)動(dòng)脈注射ADM組(n=12),側(cè)腦室注射ADM組(n=12)。采用線栓法制成大鼠大腦中動(dòng)脈缺血再灌注(MCAO)模型,阻斷血流2h進(jìn)行再灌注。缺血再灌注各組大鼠在缺血2h再灌注4h時(shí),參考Bederson等的5分制評(píng)分標(biāo)準(zhǔn)對(duì)其進(jìn)行神經(jīng)功能缺損評(píng)分,評(píng)分后分別制備石蠟切片和冰凍切片,TTC染色測(cè)定梗死體積,HE染色光鏡下觀察大鼠大腦皮質(zhì)、海馬的組織學(xué)變化,TUNEL法檢測(cè)神經(jīng)元凋亡,免疫組織化學(xué)法(SABC法)檢測(cè)ADM陽(yáng)性細(xì)胞表達(dá),原位雜交法檢測(cè)ADM mRNA
[Abstract]:Objective adrenomedullin (ADM) is a vasoactive polypeptide consisting of 52 amino acids. It belongs to the calcitonin gene-related peptide (CGRP) family and is related to ischemia-reperfusion injury in various tissues. Some scholars have confirmed the expression of ADM in the central nervous system, which is related to cerebral ischemia reperfusion injury. There are also reports of contrary views. So we chose the rat model of focal cerebral ischemia-reperfusion, using immunohistochemistry, in situ hybridization histochemistry, reverse transcription-polymerase chain reaction (RT-PCR), and so on. To verify the expression of ADM and ADM mRNA in the brain of normal rats. To observe the expression of ADM and ADM mRNA in the brain of rats with focal ischemia-reperfusion. To investigate the effects of ADM on neuronal apoptosis, infarct volume and Egr-1 mRNA in focal ischemia-reperfusion rats, and to further study the role of ADM in focal ischemia-reperfusion brain injury. Methods 186 healthy male SD rats, 200-250 g, were provided by Experimental Animal Center of China Medical University. They were randomly divided into 4 groups: normal group (nnm12), sham-operation group (NN12), focal cerebral ischemia 2h reperfusion group (NN18), focal cerebral ischemia 2h reperfusion 2 h group (NN18), 4h group (nong18), 22h group (nnm54), 46h group (nm18), 70h group (nm18), 118h group (N18), 166h group (NN18). Among them, 22 h reperfusion group was divided into three groups: femoral vein injection group (ADM 12), internal carotid artery injection group (ADM group 12), lateral ventricle injection group (ADM group 12). The middle cerebral artery (MCA) ischemia reperfusion (MCAO) model was established by the method of thread embolization, and the blood flow was blocked for 2 h for reperfusion. After ischemia 2 h and reperfusion 4 h after ischemia, the rats were evaluated for neurological impairment by referring to the 5-point scoring standard of Bederson et al. After scoring, paraffin sections and frozen sections were prepared, and the infarct volume was measured by TTC staining. Histological changes of cerebral cortex and hippocampus of rats were observed under HE staining light microscope, neuronal apoptosis was detected by TUNEL method, expression of ADM positive cells was detected by immunohistochemistry (SABC method), ADM mRNA was detected by in situ hybridization.
【學(xué)位授予單位】：中國(guó)醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2005
【分類號(hào)】：R363

【共引文獻(xiàn)】

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1 畢國(guó)榮;張輝;周慧杰;張賀敏;海虹;方秀斌;;大鼠局灶性缺血再灌注后大腦皮質(zhì)ADM及其MRNA的表達(dá)(英文)[J];Neuroscience Bulletin;2005年06期

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本文編號(hào)：2344564