【摘要】：目的 本實(shí)驗(yàn)探討卵泡抑素、激活素A與骨形成蛋白-4在正常大鼠和缺血缺氧性腦損傷大鼠腦組織皮層和海馬區(qū)的表達(dá)規(guī)律及意義。綜合卵泡抑素與兩者的作用機(jī)制，找到二者信號(hào)的交叉點(diǎn)，為臨床指導(dǎo)神經(jīng)重建提供實(shí)驗(yàn)依據(jù)。 方法 將同期受孕60只SD（Sprague dawley）大鼠分為正常組和模型組各30只，每組按照胎鼠發(fā)育時(shí)間隨機(jī)分為胚胎期8.5天、胚胎期13天、胚胎期18天，出生后3天、出生后7天、出生后30天6個(gè)亞組（即E8.5組，E13組，E18組，p3組，P7組，P30組六個(gè)亞組），胚胎期和出生后分別采用不同造模方法建立腦缺血缺氧模型，應(yīng)用SABC免疫組化方法觀察檢測(cè)生長(zhǎng)發(fā)育不同時(shí)期正常大鼠和腦缺血缺氧后大鼠卵泡抑素（Follistatin，F(xiàn)S），，激活素A（Activin A，ACT A)與骨形成蛋白-4（Bone morphogenic protein-4，BMP-4）蛋白的表達(dá)情況。通過(guò)RT-PCR方法，檢測(cè)生長(zhǎng)發(fā)育不同時(shí)期正常大鼠和腦缺血缺氧后大鼠卵泡抑素、激活素A與骨形成蛋白-4基因的表達(dá)情況。 結(jié)果 HE染色結(jié)果顯示缺血缺氧后的腦組織神經(jīng)元壞死，部分細(xì)胞核核固縮深染，局限性神經(jīng)元數(shù)目減少，神經(jīng)纖維排列紊亂，呈急性缺血改變。神經(jīng)功能檢測(cè)發(fā)現(xiàn)神經(jīng)評(píng)分隨著大鼠年齡的增長(zhǎng)而增高（P0.01），即年齡越大神經(jīng)損傷越加重。通過(guò)免疫組織化學(xué)方法顯示正常組和模型組隨著大鼠發(fā)育的不同時(shí)期腦組織皮層和海馬區(qū)的卵泡抑素和激活素A表達(dá)量逐漸減少（P0.01），BMP-4在胚胎期幾乎不表達(dá)，出生后30天呈現(xiàn)低表達(dá)（P0.01）。RT-PCR方法檢測(cè)顯示缺血缺氧后模型組卵泡抑素、激活素A與骨形成蛋白-4的表達(dá)量基本都略高于正常組（P0.01），尤其以卵泡抑素和激活素A的表達(dá)增高最為顯著，海馬區(qū)三者的表達(dá)不如皮層區(qū)增高明顯。 結(jié)論 大鼠腦缺血缺氧模型制造成功。卵泡抑素、激活素A與骨形成蛋白-4的表達(dá)與生長(zhǎng)發(fā)育和年齡密切相關(guān)，隨著大鼠年齡的增長(zhǎng)，卵泡抑素和激活素A的表達(dá)逐漸減少，而骨形成蛋白-4的表達(dá)則在成年后明顯。腦缺血缺氧損傷可誘導(dǎo)卵泡抑素、激活素A和骨形成蛋白-4的高表達(dá)，是神經(jīng)損傷的保護(hù)性因子，且卵泡抑素和激活素A是生長(zhǎng)發(fā)育早期的保護(hù)性因子。但是BMP-4的表達(dá)增高不如卵泡抑素和激活素A明顯，推測(cè)卵泡抑素的主要功能可能是作為激活素A的抑制劑而不是骨形成蛋白-4的配體來(lái)調(diào)控神經(jīng)發(fā)育。
[Abstract]:Objective to investigate the expression and significance of follicular somatostatin, activin A and bone morphogenetic protein-4 in cerebral cortex and hippocampus of normal rats and rats with ischemic and anoxic brain injury. To find the intersection of the two signals, and to provide experimental basis for clinical guidance of nerve reconstruction. Methods 60 SD (Sprague dawley) rats were divided into normal group (n = 30) and model group (n = 30). According to the development time of fetal rats, each group was randomly divided into embryonic stage (8.5 days), embryonic stage (13 days), embryonic stage (18 days) and postbirth (3 days). Cerebral ischemia and hypoxia models were established in 6 subgroup groups (E8.5 group, E13 group, E18 group, p3 group, P7 group, P30 group) 7 days after birth and 30 days after birth. SABC Immunohistochemical method was used to detect Follistatin,FS, Activin A and bone morphogenetic protein-4 (Bone morphogenic protein-4, in normal rats and rats after cerebral ischemia and hypoxia at different stages of growth and development. The expression of BMP-4) protein. The expression of follicle somatostatin, activin A and bone morphogenetic protein-4 gene in normal rats and rats after cerebral ischemia and hypoxia at different stages of growth and development were detected by RT-PCR. Results the results of HE staining showed that after ischemia and hypoxia, the neurons of brain tissue were necrotic, some of the nucleus was stained deeply, the number of localized neurons decreased, and the arrangement of nerve fibers was disordered, showing acute ischemic changes. Nerve function test showed that the nerve score increased with the increase of age (P 0.01), that is to say, the older the age, the more serious the nerve injury. The expression of follicle somatostatin and activin A in cerebral cortex and hippocampus of normal group and model group decreased gradually with the development of rats at different stages of development (P01), but BMP-4 was almost not expressed in embryonic stage. The expression of follicle somatostatin, activin A and bone morphogenetic protein-4 in the model group was slightly higher than that in the normal group after ischemia and hypoxia (P01). The expression of follicle somatostatin, activin A and bone morphogenetic protein-4 in the model group was slightly higher than that in the normal group (P 0.01). In particular, the expression of follicle somatostatin and activin A was the most significant, and the expression of follicle somatostatin and activin A in hippocampus was not as high as that in cortical area. Conclusion the rat model of cerebral ischemia and hypoxia is successful. The expression of follicle somatostatin, activin A and bone morphogenetic protein-4 was closely related to growth, development and age. With the increase of age, the expression of follicle somatostatin and activin A decreased gradually. The expression of bone morphogenetic protein-4 was obvious in adulthood. Cerebral ischemia and hypoxia injury can induce the high expression of follicle somatostatin, activin A and bone morphogenetic protein-4, which is the protective factor of nerve injury, and follicle somatostatin and activin A are protective factors in the early stage of growth and development. However, the increased expression of BMP-4 is not as obvious as that of follicular somatostatin and activin A. it is speculated that the main function of folliculatostatin may be to regulate nerve development as an inhibitor of activin A rather than as a ligand of bone morphogenetic protein-4.
【學(xué)位授予單位】：遼寧醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R743

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 王紅星;徐冬晨;王彤;;中樞神經(jīng)再生策略:增強(qiáng)神經(jīng)因子介導(dǎo)的再生環(huán)境“允許”作用[J];中國(guó)康復(fù)醫(yī)學(xué)雜志;2008年04期

2 方琳;劉永茂;葛敬巖;劉海巖;臺(tái)桂香;柳忠輝;;激活素結(jié)合蛋白阻斷激活素誘導(dǎo)雞胚神經(jīng)節(jié)神經(jīng)突起生長(zhǎng)作用及其機(jī)制研究[J];中風(fēng)與神經(jīng)疾病雜志;2007年04期

3 陳穎;徐苓;;激活素-抑制素-卵泡抑素系統(tǒng)研究進(jìn)展[J];生殖醫(yī)學(xué)雜志;2007年04期

4 薄濤,嚴(yán)超英,霍淑芳,李巍;結(jié)扎孕鼠雙側(cè)子宮動(dòng)脈復(fù)制圍產(chǎn)期缺氧缺血性腦損傷動(dòng)物模型[J];中風(fēng)與神經(jīng)疾病雜志;2001年01期

5 張濤;王景周;;成體腦內(nèi)神經(jīng)發(fā)生和腦缺血對(duì)其影響[J];中華神經(jīng)醫(yī)學(xué)雜志;2009年02期

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