【摘要】： 目的：研究大鼠視網(wǎng)膜急性缺血中視網(wǎng)膜節(jié)細胞(Retinal ganglion cells, RGCs)死亡時腦紅蛋白(Neuroglobin, Ngb)的表達。 方法：實驗研究。成年SD大鼠40只,2-3月齡,體重220-240g,雌性,采用特異性結(jié)扎大鼠視網(wǎng)膜動脈的方法造成視網(wǎng)膜急性缺血模型,按照結(jié)扎時間不同(0、15、30、60 min)分為4組,每組10只大鼠,均以左眼為實驗眼。每組隨機選取3只大鼠視網(wǎng)膜用于免疫印跡法檢測,取4只大鼠的視網(wǎng)膜分別做熒光金(Fluorogold, FG)逆行標記節(jié)細胞計數(shù)及免疫組化染色熒光強度分析,3只大鼠做免疫電子顯微鏡觀察。所有數(shù)據(jù)均采用單因素方差分析進行統(tǒng)計學比較,采用LSD-t檢驗進行兩兩比較。 結(jié)果：視網(wǎng)膜完全結(jié)扎15min時,視網(wǎng)膜中Ngb表達最高(P=0.000),之后逐漸降低,到結(jié)扎30min時,Ngb表達接近正常(P=0.728),此時與0min組比較RGCs數(shù)量開始減少(P=0.011),結(jié)扎60min時,Ngb的表達已明顯低于Omin組(P=0.001),與Omin組比較RGCs數(shù)量進一步減少(P=0.000)。在視網(wǎng)膜各層中,Ngb的表達以RGCs層為最高,其次為內(nèi)叢狀層與外叢狀層。RGCs中Ngb主要分布在胞質(zhì),結(jié)扎30min時線粒體外室和線粒體嵴也發(fā)現(xiàn)有Ngb表達,而內(nèi)核層及外核層細胞的胞質(zhì)內(nèi)始終未見Ngb的表達。 結(jié)論：Ngb在大鼠RGCs急性缺血死亡時表達快速升高,主要表達于RGCs的胞質(zhì)內(nèi),與RGCs的生存狀態(tài)關(guān)系密切。
[Abstract]:Aim: to study the expression of (Neuroglobin, Ngb) in rat retinal ganglion cells (RGCs) during (Retinal ganglion cells, RGCs) death in acute retinal ischemia. Methods: experimental study. 40 adult SD rats, weighing 220-240 g and weighing 220-240 g, were divided into 4 groups according to different ligation time (0 ~ 153030 ~ 60 min), each group of 10 rats with left eye as experimental eyes. The retina of each group was randomly selected for immunoblotting assay. The retinal cells of 4 rats were stained with fluorescent gold (Fluorogold, FG) retrograde labeled ganglion cells, and the fluorescence intensity of immunohistochemical staining was analyzed. 3 rats were observed by immunoelectron microscopy. All data were compared by single factor ANOVA and LSD-t test. Results: when the retina was completely ligated with 15min, the expression of Ngb in the retina was the highest (P0. 000), and then decreased gradually. By the time of ligation of 30min, the expression of NGB was close to normal (Pf0.728), and the number of RGCs began to decrease (P0. 011) compared with 0min group. The expression of NGB in 60min ligation group was significantly lower than that in Omin group (P0. 001), and the number of RGCs in Omin group was further decreased compared with Omin group (P0. 000). In all layers of retina, the expression of NGB was highest in RGCs layer, followed by Ngb in inner plexiform layer and outer plexiform layer. Ngb expression was also found in mitochondria outer chamber and mitochondrial crest during ligation of 30min. However, no expression of Ngb was found in the cytoplasm of the inner and outer nuclear layer. Conclusion the expression of% NGB was rapidly increased during the acute ischemic death of RGCs in rats, mainly in the cytoplasm of RGCs, which was closely related to the survival state of RGCs.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R774.1

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