本文關(guān)鍵詞： 視神經(jīng)/損傷 視網(wǎng)膜神經(jīng)節(jié)細(xì)胞 GFAP 時(shí)間相關(guān)性　出處：《華中科技大學(xué)》2006年碩士論文　論文類型：學(xué)位論文

【摘要】： 【研究背景】視神經(jīng)挫傷是法醫(yī)學(xué)活體鑒定和臨床眼科中一種常見的眼外傷,往往給傷者帶來不可逆轉(zhuǎn)的視功能損害,正確認(rèn)識(shí)和評(píng)價(jià)這種損傷對(duì)視功能的影響是臨床和法醫(yī)工作者的重要任務(wù)。神經(jīng)膠質(zhì)原纖維酸性蛋白(glial fibrillary acid protein,GFAP)是構(gòu)成細(xì)胞骨架中間絲的主要成分,主要存在于中樞神經(jīng)系統(tǒng)的星形膠質(zhì)細(xì)胞中[1]。神經(jīng)系統(tǒng)損傷時(shí)膠質(zhì)細(xì)胞功能的改變常表現(xiàn)為膠質(zhì)細(xì)胞免疫反應(yīng)性的改變。在許多神經(jīng)變性疾病均發(fā)現(xiàn)GFAP免疫反應(yīng)性的改變,它被認(rèn)為是中樞神經(jīng)損傷的靈敏指示劑。同時(shí),一系列的動(dòng)物模型研究證實(shí),視神經(jīng)損傷后視功能下降的病理基礎(chǔ)是以視網(wǎng)膜神經(jīng)節(jié)細(xì)胞(Retinal ganglion cells, RGCs)的繼發(fā)性喪失為主[2]。所以,正確認(rèn)識(shí)視神經(jīng)損傷后病理變化的特點(diǎn)就成了本研究的主要內(nèi)容。 【目的】本實(shí)驗(yàn)通過建立大鼠視神經(jīng)夾挫傷模型,觀察傷后RGCs的形態(tài)學(xué)改變及Müller細(xì)胞合成的GFAP的變化,為研究視神經(jīng)損傷后法醫(yī)學(xué)鑒定提供分子病理學(xué)基礎(chǔ)。 【材料與方法】Wistar成年大鼠45只,分為7組,每組動(dòng)物5只,參照文獻(xiàn)的方法,制成視神經(jīng)夾挫傷動(dòng)物模型,傷后各組分別飼養(yǎng)存活1d、3d、5d、7d、9d、14d、21d,同期行灌流固定及眼球取材固定切片。光鏡下觀察視網(wǎng)膜神經(jīng)節(jié)細(xì)胞的改變,免疫組化方法檢測(cè)不同時(shí)間RGCs表達(dá)GFAP的水平,對(duì)視網(wǎng)膜神經(jīng)節(jié)細(xì)胞及GFAP表達(dá)變化的規(guī)律與損傷時(shí)間的關(guān)系進(jìn)行分析。 【結(jié)果】視神經(jīng)損傷后誘導(dǎo)視網(wǎng)膜神經(jīng)節(jié)細(xì)胞喪失,視神經(jīng)損傷后1d,可見少量RGCs開始潰變死亡,3d時(shí)RGCs開始大量減少,7d時(shí)減少約36%,14d時(shí)減少約44%,21d時(shí)減少約52%。7d之前細(xì)胞數(shù)目快速減少,14d后呈慢速減少,14d到21d內(nèi)細(xì)胞丟失數(shù)目?jī)H為7.43%,而正常組RGCs沒有任何明顯變化。視神經(jīng)損傷后1d視網(wǎng)膜GFAP染色與正常大鼠視網(wǎng)膜相似;傷后3d,視網(wǎng)膜神經(jīng)纖維層和節(jié)細(xì)胞層有增多的GFAP陽性染色,內(nèi)網(wǎng)狀層、內(nèi)核層、外網(wǎng)狀層可見線條狀GFAP陽性著色;傷后7d,節(jié)細(xì)胞層及內(nèi)、外界膜之間的GFAP陽性染色進(jìn)一步加深、密集,并達(dá)到高峰;傷后9d,GFAP表達(dá)下降,但下降緩慢,已趨于平緩,14d后下降至接近正常水平。 【結(jié)論】從視網(wǎng)膜病理改變結(jié)果看出:與之前研究結(jié)果相比,隨著致傷強(qiáng)度增加,RGC丟失加重,說明不同的致傷強(qiáng)度造成不同程度的視神經(jīng)損傷,致傷強(qiáng)度和損傷程度成負(fù)相關(guān)。實(shí)驗(yàn)發(fā)現(xiàn)視神經(jīng)損傷后3d時(shí)RGCs開始大量喪失,7d內(nèi)迅速減少,到14d時(shí)已達(dá)44%,14d后喪失速度明顯放慢,21d后幾乎無明顯變化,提示在視神經(jīng)外傷后,RGCs的喪失與時(shí)間有明顯相關(guān)性,這與國外學(xué)者Berkelaar等[3]的研究結(jié)果相一致。視神經(jīng)夾挫傷使視網(wǎng)膜GFAP免疫反應(yīng)性于傷后3d增加,7d達(dá)到高峰,表明視神經(jīng)損傷引起視網(wǎng)膜膠質(zhì)細(xì)胞反應(yīng)性增高,隨時(shí)間進(jìn)展又逐漸下降,于14d后降至正常水平,這與受損RGCs的潰變、死亡率的逐漸增高是一致的。同時(shí)實(shí)驗(yàn)觀察到GFAP的表達(dá)部位主要集中在RGCs的胞膜及其纖維的周圍,表明GFAP蛋白與RGCs的存活數(shù)量密切相關(guān),隨著GFAP表達(dá)的減弱,受損RGCs不斷潰變和死亡,表達(dá)也隨之減弱。綜上所述,視神經(jīng)損傷后直接機(jī)械損傷作用導(dǎo)致視功能迅速下降,繼發(fā)性的RGCs喪失和軸突的潰變導(dǎo)致視功能的惡化并顯示出時(shí)間相關(guān)性;GFAP蛋白表達(dá)水平也表現(xiàn)出一定的時(shí)間相關(guān)性,其表達(dá)強(qiáng)弱與RGCs存活數(shù)量多少也密切相關(guān)。這些為正確了解視神經(jīng)損傷后視功能變化的病理因素和預(yù)后評(píng)估提供了重要的理論基礎(chǔ),也為視神經(jīng)挫傷的鑒定時(shí)機(jī)提供了依據(jù)。
[Abstract]:[background] optic nerve contusion is a common ocular trauma in forensic science in vivo identification and Clinical Ophthalmology, often to bring injured visual function damage is irreversible, affect the correct understanding and evaluation of the damage of visual function is an important task in clinical and forensic medicine. Glial fibrillary acidic protein (glial fibrillary acid protein. GFAP) is the main component of cytoskeletal intermediate filaments, injury of glial cell function of [1]. neural system mainly exists in the central nervous system in astrocytes glial cell changes are often immune reactive changes. In many neurodegenerative diseases were found GFAP immunoreactivity changes, it is considered to be sensitive the indicator of central nervous system injury. At the same time, a series of studies in animal models confirmed that the pathological basis of optic nerve damage visual function decline in the retina of God The secondary loss of Retinal ganglion cells (RGCs) is mainly [2].. Therefore, correctly recognizing the characteristics of pathological changes after optic nerve injury has become the main content of this research.
[Objective] optic nerve crush model in rats was established by this experiment, the change of RGCs morphology and M ller cell GFAP synthesis was observed after injury of optic nerve injury in forensic medicine provides the molecular pathologic basis.
[materials and methods] 45 adult Wistar rats were divided into 7 groups, each group of animal 5, according to the literature method, made of optic nerve crush injury animal model, each group were reared 1D 3D, 5D, survival, 7d, 9D, 14d, 21d, underwent perfusion fixation and eyeball were fixed slice. Under the light microscope observation on retinal ganglion cell changes, immunohistochemical method to detect the expression level of GFAP RGCs in different time, we analyzed the relationship between law and time of injury on the expression of retinal ganglion cells and GFAP.
[results] after optic nerve injury induced by retinal ganglion cell loss after optic nerve injury, 1D, a small amount of RGCs degeneration death, 3D RGCs began to decrease by about 36% 7d, reduced by about 44% 14d, about 52%.7d before the cell number decreased rapidly decreased 21d, 14d after a slow decrease, 14d 21d cell loss is only 7.43%, and no significant change in normal group. RGCs of 1D after optic nerve injury of retina GFAP staining and normal rat retina; 3D after injury, the retinal nerve fiber layer and ganglion cell layer with positive staining of GFAP increased, the inner plexiform layer, inner nuclear layer, outer plexiform layer visible line GFAP staining; 7d after injury, and the ganglion cell layer, GFAP positive staining between the outside membrane further, dense, and reached the peak at 9D after injury; and the expression of GFAP decreased, but the decline has tended to slow, gentle, after 14d decreased to normal water Ping.
[Conclusion] from the retinal pathological changes showed that compared with the previous research results, with the injury intensity increases, the loss of RGC increased, the intensity of injuries caused by different show different degrees of optic nerve injury, negatively related to the intensity of injuries and damage degree. The experimental results showed that optic nerve injury after 3D RGCs lost a lot, quickly reduce 7d, 14d has reached 44%, after the loss of 14d slowed down significantly, 21d almost no obvious change, suggesting that in the optic nerve after injury, loss of RGCs have significant correlation with time, consistent with the results of the research of foreign scholar Berkelaar [3]. Optic nerve crush GFAP immunoreactive retinal the increase in 3D after injury, reached the peak at 7d, that caused the increase of retinal glial cell reaction of optic nerve injury, over time and gradually decreased, after 14d decreased to normal level, which was associated with impaired RGCs degeneration, mortality by Gradually increase is consistent. At the same time around the observed expression site of GFAP mainly focus on the cell membrane and RGCs fiber, GFAP protein and RGCs showed that the number of survival is closely related with the decreased expression of GFAP, RGCs has damaged the degeneration and death, expression is weaken. To sum up, resulting in direct effect the mechanical damage of visual function after optic nerve injury decreases rapidly, secondary loss of RGCs and axonal degeneration leads to the deterioration of visual function and displays the time correlation; the expression level of GFAP protein also showed a certain time correlation, the expression intensity of RGCs and survival is closely related to the number. These provide an important theoretical basis of pathology the factors and prognosis of optic nerve function changes after its injury in order to correctly understand, also provides the basis for the identification of the timing of optic nerve contusion.

【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：D919

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 狄靜芳,史劍波,曾耀英,徐錦堂;視神經(jīng)損傷后視網(wǎng)膜細(xì)胞凋亡的研究[J];暨南大學(xué)學(xué)報(bào)(自然科學(xué)與醫(yī)學(xué)版);1999年04期

2 李立新;反應(yīng)性星形膠質(zhì)化的形成機(jī)制及研究進(jìn)展[J];國外醫(yī)學(xué)(生理、病理科學(xué)與臨床分冊(cè));1996年04期

3 張?jiān)趶?qiáng),龍潔,李小玲,趙性泉;局灶性鼠腦缺血腦組織環(huán)氧酶-2的表達(dá)與細(xì)胞凋亡的關(guān)系[J];現(xiàn)代康復(fù);2000年10期

4 袁洪峰,劉少章,賀翔鴿;大鼠視神經(jīng)損傷后一氧化氮對(duì)視網(wǎng)膜功能的影響[J];現(xiàn)代康復(fù);2001年14期

5 孫安;顱腦損傷后的營養(yǎng)和攝食問題[J];現(xiàn)代康復(fù);2001年24期

6 袁洪峰,劉少章,賀翔鴿,龍正勤;大鼠視神經(jīng)損傷視網(wǎng)膜節(jié)細(xì)胞凋亡的實(shí)驗(yàn)研究[J];中國臨床康復(fù);2002年13期

7 劉成龍,靳安民,童斌輝,閔少雄,田京;脊髓損傷后組織基因表達(dá)譜的變化[J];中國臨床康復(fù);2003年17期

8 何祥,張微;中樞神經(jīng)系統(tǒng)損傷后出芽的研究進(jìn)展[J];中國臨床康復(fù);2003年25期

9 崔志利,王琳,胡丹,康軍,惠延年;神經(jīng)營養(yǎng)因子促進(jìn)視神經(jīng)夾傷后視網(wǎng)膜生長(zhǎng)相關(guān)蛋白-43表達(dá)[J];眼科新進(jìn)展;2003年02期

10 胡愛蓮,孫葆忱,李遼青,孫憲麗,劉守彬,董冬生;大鼠視神經(jīng)夾挫傷視網(wǎng)膜視神經(jīng)病理學(xué)動(dòng)態(tài)觀察及功能檢測(cè)[J];眼科;2003年06期

，

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