【摘要】：在臨床中創(chuàng)傷性腦損傷(traumatic brain injury TBI）是神經(jīng)外科常見多發(fā)疾病，由其導(dǎo)致的病死率、殘障率在各類創(chuàng)傷性損傷中比例最高。TBI后患者近、遠(yuǎn)期易并發(fā)癲癇、認(rèn)知功能障礙等并發(fā)癥，嚴(yán)重影響患者生存質(zhì)量及生活質(zhì)量。由于對TBI后的神經(jīng)細(xì)胞內(nèi)生化改變?nèi)狈θ嬲J(rèn)識，因此選擇對其有針對性的治療一直是困擾臨床的難點(diǎn)問題。其認(rèn)識理論、治療方法、療效至今仍沒有獲得突破性進(jìn)展。大量研究表明微管蛋白（tubulin）家族不但與細(xì)胞分裂、增殖、凋亡、腫瘤的發(fā)生發(fā)展密切相關(guān)，還與許多神經(jīng)系疾病以及學(xué)習(xí)記憶、認(rèn)知功能密切相關(guān)。β-tubulin廣泛表達(dá)于多種細(xì)胞的胞質(zhì)中，具有一定的組織和細(xì)胞表達(dá)特異性，β-tubulin在正常腦組織中表達(dá)量高。但國內(nèi)外尚無TBI后β-tubulin量變引起神經(jīng)元損傷與認(rèn)知功能改變之間關(guān)系的研究報道。作者擬采用建立大鼠TBI實(shí)驗(yàn)?zāi)Ｐ脱芯縏BI后腦組織中β-tubulin分子的變化規(guī)律。 目的：探究創(chuàng)傷性腦損傷神經(jīng)損傷的機(jī)理，初步了解細(xì)胞骨架蛋白成分之一的β-微管蛋白與創(chuàng)傷性腦損傷之間的關(guān)系以及損傷后不同時間的表達(dá)量和變化趨勢。模擬中度創(chuàng)傷性腦損傷制作TBI大鼠模型，旨在從蛋白表達(dá)變化水平角度研究TBI后神經(jīng)元損傷發(fā)生的分子機(jī)制，期望通過動物實(shí)驗(yàn)提高臨床診治創(chuàng)傷性腦損傷的水平，探索有效腦保護(hù)措施的理論依據(jù)。 方法：選用共70只健康SD大鼠，清潔等級，雄性，體重（220±20g）。動物編號后采用隨機(jī)法劃分為正常對照組（10只）和TBI模型組（60只）；運(yùn)用頭顱瞬間側(cè)向旋轉(zhuǎn)致傷裝置制作大鼠TBI模型。按創(chuàng)傷后動物處死時間點(diǎn)劃分為6h、12h、24h、72h、7d、14d6個亞組（每組10只）。在各規(guī)定時間點(diǎn)處死動物模型，灌注固定腦組織，采集腦組織制作石蠟切片，采用免疫組織化學(xué)（immunohistochemistry，IHC）技術(shù)檢測β-微管蛋白在大鼠海馬和皮層組織的分布情況，研究其在腦損傷后腦組織不同部位的表達(dá)量及變化規(guī)律。將陽性表達(dá)部位圖片應(yīng)用圖像處理軟件轉(zhuǎn)化為灰度值，統(tǒng)計(jì)其灰度值，實(shí)驗(yàn)數(shù)據(jù)用均數(shù)±標(biāo)準(zhǔn)差（x±SD）表示，應(yīng)用SPSS17.0統(tǒng)計(jì)軟件包進(jìn)行數(shù)據(jù)分析，采用u檢驗(yàn)，設(shè)定統(tǒng)計(jì)學(xué)差異顯著水平為P0.05。 結(jié)果：免疫組化檢測結(jié)果顯示在大鼠海馬區(qū)、齒狀回及大腦皮層下神經(jīng)細(xì)胞中β-tubulin廣泛分布在的胞漿、軸突中。對照組的β-tubulin陽性表達(dá)量較TBI模型組表達(dá)量高，海馬區(qū)表達(dá)量高于皮層區(qū)。創(chuàng)傷后不同時間點(diǎn)表達(dá)量不同，在TBI后6h模型組β-tubulin表達(dá)陽性量少，為最低值（p 0.05），隨著創(chuàng)傷時間的推移延長，其表達(dá)量逐漸增加。在皮層組織中恢復(fù)相對較快，至7d時表達(dá)量與正常對照組基本相同。與正常對照組差異比較無統(tǒng)計(jì)學(xué)意義（p0.05）。但在海馬組織區(qū)在損傷后14d組β-tubulin表達(dá)量與對照組接近，，與正常對照組比較，差異顯著性無統(tǒng)計(jì)學(xué)意義（p0.05）。 結(jié)論：在創(chuàng)傷性腦損傷模型腦中，大鼠神經(jīng)元中β-tubulin表達(dá)量早期明顯下降，隨創(chuàng)傷時間推移β-tubulin表達(dá)量逐漸升高。創(chuàng)傷對腦組織的不同部位影響不同，不同區(qū)域的神經(jīng)細(xì)胞恢復(fù)能力不同，皮層區(qū)恢復(fù)較海馬區(qū)快，損傷后7dβ-tubulin基本恢復(fù)正常水平。海馬區(qū)至損傷后14d時β-tubulin表達(dá)量基本恢復(fù)至對照組水平。推論β-tubulin參與神經(jīng)損傷與修復(fù)的過程，有必要繼續(xù)進(jìn)一步深入研究其變化的機(jī)理、機(jī)制，并進(jìn)行相應(yīng)的藥物干預(yù)研究。
[Abstract]:Traumatic brain injury (TBI) is a common multiple of neurosurgery. The mortality and disability rate are the highest among all kinds of traumatic injury. The near-term and long-term complications of patients with TBI, such as epilepsy, cognitive impairment, and the like, seriously affect the quality of life and quality of life of patients. Because of the lack of comprehensive understanding of the biochemical changes in the nerve cells after TBI, the selection of the targeted therapy has been a difficult problem for clinical. The cognitive theory, the treatment method and the curative effect have not yet achieved breakthrough progress. A large number of studies have shown that the tubuin family not only is closely related to cell division, proliferation, apoptosis, and development of the tumor, but also is closely related to many nerve system diseases and learning and memory and cognitive function. Conclusion-tubelin is widely expressed in the cytoplasm of a plurality of cells, and has certain tissue and cell expression specificity, and the expression level of the antigen-tubelin in the normal brain tissue is high. However, there are no studies on the relationship between the changes of neuronal damage and cognitive function after TBI in the home and abroad. The authors proposed to establish a rat TBI experimental model to study the changes of the P-tubuin molecule in the brain tissue after TBI. Objective: To study the mechanism of the nerve injury of traumatic brain injury, and to understand the relationship between the antigen-microtubule protein and the traumatic brain injury of one of the components of the cellular skeleton protein and the expression and change of the time after the injury. Potential: To study the molecular mechanism of the post-traumatic brain injury of TBI from the level of protein expression change, and to explore the theory of effective brain protective measures in order to improve the level of clinical diagnosis and treatment of traumatic brain injury by animal experiments. Methods:70 healthy SD rats, clean grade, male, body weight (220%2) were selected 0 g). The animal number was divided into the normal control group (10 rats) and the TBI model group (60 rats) by using the random method, and the rats were made of TB using the head instant lateral rotation injury device. I model. The time points were divided into 6 h,12 h,24 h,72 h,7 d,14 d6 subgroups according to the post-traumatic animal sacrifice time point (each group 1 (0). The animal model was sacrificed at the specified time points, the brain tissue was fixed by perfusion, the paraffin section was made from the brain tissue, and the score of the antigen-microtubule protein in the hippocampus and the cortex of the rat was detected by using the immunohistochemistry (IHC) technique. The expression of different parts of brain tissue after brain injury and its change after brain injury The image of the positive expression part is transformed into the gray value by the image processing software, the gray value of the image is counted, the average standard deviation (x-SD) of the average number of the experimental data is used for data analysis, the data analysis is carried out by using the SPSS17.0 statistical software package, and the statistical difference is set to P0 by using the u test. .05. Results: The results of immunohistochemistry showed that in the rat hippocampus, dentate gyrus and the neurons in the cerebral cortex were widely distributed in the cells. The positive expression of P-tubuin in the control group was higher than that of the TBI model group, and the expression of the hippocampal region was higher than that of the TBI model group. It was higher than that in cortex area. The expression was different at different time points after trauma. The expression of antigen-tubiulin in the 6-h model group after TBI was small and the lowest value (p.05), and the expression was prolonged with the time of trauma. The amount gradually increased. The recovery was relatively fast in the cortical tissue, and the amount of expression was similar to that of the normal control at 7 d. The group was basically the same. The difference with the normal control group was not statistically significant (p 0.05). However, in the hippocampus, the expression of e-tubuin was close to the control group in 14 days after the injury, and there was no significant difference in the difference between the control group and the control group (p 0.05). Conclusion: In the brain of traumatic brain injury model, the expression of l-tubuin in the rat's neurons was significantly decreased, and with the time of traumas-tubuin. The effect of trauma on different parts of brain tissue was different, and the recovery of nerve cells in different regions was different. The normal level of the expression in the hippocampus from the hippocampus to the 14-day post-injury level It is necessary to continue to study the mechanism and mechanism of the changes in the process of nerve injury and repair.
【學(xué)位授予單位】：延安大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R651.15

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 房博;賀曉生;陳延;梁明;陳曉燕;袁志海;;創(chuàng)傷性腦損傷后大鼠腦組織S100A6的表達(dá)規(guī)律及意義[J];中華神經(jīng)外科疾病研究雜志;2012年04期

2 肖計(jì)劃,夏潮涌,賈艷濱,鄭佩娥;阿爾茨海默病與血管性癡呆基底細(xì)胞微管蛋白變化研究[J];中國病理生理雜志;2001年07期

3 賀曉生,易聲禹,章翔,費(fèi)舟,張劍寧,梁景文,楊利孫;腦彌漫性軸索損傷軸索內(nèi)鈣超載及鈣拮抗劑的治療作用[J];中華神經(jīng)外科雜志;2000年01期

相關(guān)博士學(xué)位論文 前1條

1 馬學(xué)玲;大鼠局灶性腦缺血蛋白質(zhì)組研究及hMSCs定向移植治療腦缺血[D];吉林大學(xué);2008年

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