本文選題：YFP小鼠　切入點(diǎn)：GFP小鼠　出處：《蘭州大學(xué)》2017年碩士論文

【摘要】：中樞神經(jīng)系統(tǒng)疾病的治療和功能的恢復(fù)一直是醫(yī)學(xué)界的難題。眼內(nèi)存在的創(chuàng)傷應(yīng)激、病理機(jī)制、炎癥反應(yīng)等與腦和脊神經(jīng)中十分相似,是中樞神經(jīng)系統(tǒng)研究的重要模型。目前對(duì)小膠質(zhì)細(xì)胞在中樞神經(jīng)創(chuàng)傷和疾病中的作用還不能定論。本實(shí)驗(yàn)以視神經(jīng)軸突被標(biāo)記的THY1-YFP H-lin轉(zhuǎn)基因小鼠和小膠質(zhì)細(xì)胞被標(biāo)記的GFP-C57BL/6轉(zhuǎn)基因小鼠為實(shí)驗(yàn)對(duì)象,建立成體小鼠單側(cè)眼視神經(jīng)軸索鈍性損傷模型,在手術(shù)后4小時(shí)、1天、3天、5天、10天分離視神經(jīng),用共聚焦熒光顯微鏡觀察視神經(jīng)軸突退行性病變和小膠質(zhì)細(xì)胞的變化。實(shí)驗(yàn)結(jié)果顯示,視神經(jīng)損傷后退行性病變可分為三個(gè)階段,一是起始階段:損傷后4小時(shí),視神經(jīng)損傷處軸突呈斷開狀態(tài),形成膠質(zhì)瘢痕,靜息態(tài)小膠質(zhì)細(xì)胞大量出現(xiàn),其中部分激活并向受損區(qū)遷移。二是退行性病變階段:損傷后1~5天,視神經(jīng)軸突從部分念珠化到大量念珠化,說明神經(jīng)軸突已經(jīng)進(jìn)入退行性病變進(jìn)程,可以觀察到激活態(tài)小膠質(zhì)細(xì)胞大量增多并覆蓋損傷區(qū)。三是退行性病變快速惡化階段:損傷后5、10天,視神經(jīng)軸突的退行性病變持續(xù)惡化,軸突從念珠狀轉(zhuǎn)變?yōu)樗槠瑺?激活態(tài)小膠質(zhì)細(xì)胞聚集且數(shù)量保持穩(wěn)定,少數(shù)小膠質(zhì)細(xì)胞有向靜息態(tài)轉(zhuǎn)變的趨勢(shì)。實(shí)驗(yàn)中也發(fā)現(xiàn)小鼠單側(cè)眼損傷后,其對(duì)側(cè)眼中也出現(xiàn)了大量的小膠質(zhì)細(xì)胞,這些細(xì)胞雖然被激活但并沒有對(duì)正常的神經(jīng)軸突產(chǎn)生影響。本實(shí)驗(yàn)的結(jié)果表明成體小鼠視神經(jīng)損傷后軸突的退行性病變持續(xù)惡化并伴隨著小膠質(zhì)細(xì)胞的激活和增多,說明小膠質(zhì)細(xì)胞與視神經(jīng)的退行性病變密切關(guān)聯(lián)。本實(shí)驗(yàn)旨在通過視神經(jīng)損傷模型了解中樞神經(jīng)退行性病變和小膠質(zhì)細(xì)胞的作用,為這類疾病的研究提供新的思路。
[Abstract]:The treatment and recovery of central nervous system diseases has been a difficult problem in the medical field.Traumatic stress, pathological mechanism and inflammatory reaction in the eye are very similar to those in the brain and spinal nerve, and are important models for the study of the central nervous system (CNS).The role of microglia in central nervous trauma and disease is uncertain.In this experiment, THY1-YFP H-lin transgenic mice labeled with optic nerve axons and GFP-C57BL/6 transgenic mice labeled with microglia were used as experimental objects to establish the model of unilateral optic nerve axonal blunt injury in adult mice.The optic nerve was separated from the optic nerve 4 hours a day, 3 days, 5 days and 10 days after operation. The changes of axonal degeneration and microglia of optic nerve were observed by confocal fluorescence microscope.The experimental results show that the degenerative lesions after optic nerve injury can be divided into three stages. One is the initial stage: 4 hours after the injury, the axons of the optic nerve are disconnected, the glial scar is formed, and the resting microglia appear in large numbers.Some of them were activated and migrated to the damaged area.Second, degenerative lesion stage: the axons of optic nerve changed from partial rosary to large amount of rosary on the 1st and 5th day after injury, which indicated that the axons of the nerve had entered the process of degenerative lesion, and the active microglia could be observed to increase a lot and cover the injured area.The third is the stage of rapid deterioration of degenerative lesions: after 5 to 10 days of injury, the degeneration of optic nerve axons continued to deteriorate, the axons changed from beads to fragments, the activated microglia gathered and the number of activated microglia remained stable.A small number of microglial cells have a tendency to move to a resting state.It was also found that a large number of microglia appeared in the contralateral eyes after unilateral eye injury. Although these cells were activated, they did not affect the normal axons.The results of this study showed that the axonal degeneration of adult mice after optic nerve injury continued to deteriorate with the activation and increase of microglia indicating that microglia were closely related to optic nerve degenerative lesions.The purpose of this study was to understand the role of neurodegenerative lesions and microglia in central nervous system by optic nerve injury model, and to provide new ideas for the study of these diseases.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R741

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