發(fā)布時間：2019-06-04 15:14

【摘要】：背景：從解剖上看，視神經(jīng)屬于周圍神經(jīng)系統(tǒng)（PNS），但如果從其發(fā)育過程中所表現(xiàn)出來的特點，以及所具有的結(jié)構(gòu)特征和功能特性來看，視神經(jīng)與中樞神經(jīng)系統(tǒng)（CNS）的神經(jīng)更為相似，所以常被視作一種特殊分化的中樞神經(jīng)。因此，與CNS的神經(jīng)元類似，，RGCs損傷后的再生也是極為困難的，這是眼科臨床上外傷性視神經(jīng)疾病和青光眼等引起不可逆盲的主要原因。 近年來研究表明：CNS難以有效的再生，神經(jīng)元自身的性質(zhì)并不是最為主要的因素，損傷導(dǎo)致神經(jīng)周圍形成的具有強烈抑制作用的微環(huán)境是更為重要的原因。再生軸突往往由于這種微環(huán)境的存在，在出芽的初始階段就發(fā)生了萎縮，最終導(dǎo)致了神經(jīng)再生的失敗。目前，微環(huán)境在神經(jīng)再生研究領(lǐng)域的關(guān)注度正在快速增加。 雄激素是近年來研究得較多的一種性激素。除了具有促進生殖系統(tǒng)發(fā)育和維持第二性征的作用外，還具有腦保護功能。然而，關(guān)于雄激素是否對視神經(jīng)具有保護作用，以及是否通過調(diào)節(jié)神經(jīng)再生微環(huán)境來發(fā)揮作用報道不多。 目的：1.建立誘導(dǎo)分化后的大鼠RGC-5細胞神經(jīng)突起損傷模型；2.觀察二氫睪酮（DHT）對神經(jīng)突起損傷后再生的作用；3.檢測二氫睪酮（DHT）對損傷局部微環(huán)境中RhoA和Neuritin表達的影響。 方法：1.用Na2S2O4制備大鼠RGC-5細胞神經(jīng)突起氧糖剝奪/復(fù)氧損傷模型；2.用不同濃度的DHT（1、10和100nmol/L）對損傷模型進行干預(yù)，MTT法檢測細胞存活率，光學(xué)顯微鏡下觀察細胞神經(jīng)突起數(shù)量和長度，RT-PCR反應(yīng)和Western blot法檢測細胞RhoA和NeuritinmRNA和蛋白的表達水平。 結(jié)果：1.5mmol/L的Na2S2O4為制備RGC-5細胞神經(jīng)突起氧糖剝奪/復(fù)氧損傷模型的最佳濃度。2.氧糖剝奪/復(fù)氧損傷后，模型組細胞存活率、神經(jīng)突起數(shù)量和長度均較正常對照組減少（P＜0.05），DHT干預(yù)后，與模型組比較，10和100nmol/L組細胞存活率和神經(jīng)突起數(shù)量增加（P＜0.05），而1nmol/L組細胞存活率和神經(jīng)突起數(shù)量差異沒有統(tǒng)計學(xué)意義（P0.05），1、10和100nmol/L DHT組神經(jīng)突起長度較模型組增加（P＜0.05）。3.氧糖剝奪/復(fù)氧損傷后，模型組RhoA mRNA和蛋白表達量均較正常對照組明顯增加（P＜0.05），而Neuritin mRNA及蛋白表達量均較正常對照組明顯減少（P＜0.05），DHT干預(yù)后，1、10和100nmol/L DHT組RhoA mRNA和蛋白表達量均較模型組減少（P＜0.05），而Neuritin mRNA及蛋白表達量均較模型組增加（P＜0.05）。 結(jié)論：RhoA和Neuritin可能是氧糖剝奪/復(fù)氧損傷后影響大鼠RGC-5細胞神經(jīng)突起再生的重要因子之一，DHT可能通過調(diào)節(jié)RhoA和Neuritin的表達，從而發(fā)揮神經(jīng)保護作用。
[Abstract]:Background: from the anatomical point of view, the optic nerve belongs to the peripheral nervous system (PNS), but if you look at the characteristics of its development, as well as the structural and functional characteristics, The optic nerve is more similar to the nerve of the central nervous system (CNS), so it is often regarded as a specially differentiated central nervous system. Therefore, similar to the neurons of CNS, the regeneration after RGCs injury is extremely difficult, which is the main cause of irreversible blindness caused by traumatic optic nerve disease and glaucoma in ophthalmic clinic. In recent years, it has been shown that CNS is difficult to regenerate effectively, and the nature of neurons is not the most important factor, and the microenvironment around the nerve is more important because of the strong inhibitory microenvironment around the nerve. Because of the existence of this microenvironment, the regenerated axons atrophy at the initial stage of budding, which leads to the failure of nerve regeneration. At present, the attention of microenvironment in the field of nerve regeneration is increasing rapidly. Androgen is a kind of sex hormone which has been studied more and more in recent years. In addition to promoting the development of the reproductive system and maintaining the secondary sexual characteristics, it also has the function of brain protection. However, there are few reports about whether androgen has protective effect on optic nerve and whether androgen plays a role by regulating nerve regeneration microenvironment. Objective: 1. The neural process injury model of rat RGC-5 cells induced by differentiation was established. To observe the effect of dihydrotestosterone (DHT) on regeneration after nerve process injury. To detect the effect of dihydrotestosterone (DHT) on the expression of RhoA and Neuritin in local microenvironment. Methods: 1. The rat model of neuronal oxygen glucose deprivation / reoxidation injury in RGC-5 cells was established by Na2S2O4. Different concentrations of DHT (1, 10 and 100nmol/L) were used to intervene the injury model. The cell survival rate was measured by MTT assay, and the number and length of neuronal processes were observed under optical microscope. The expression of RhoA, NeuritinmRNA and protein were detected by RT-PCR reaction and Western blot assay. Results: the Na2S2O4 of 1.5mmol/L was the best concentration for the establishment of neuronal oxygen glucose deprivation / reoxidation injury model in RGC-5 cells. 2. After oxygen-glucose deprivation / reoxygenation injury, the cell survival rate, the number and length of nerve processes in the model group were lower than those in the normal control group (P < 0.05), DHT), compared with those in the model group. The cell survival rate and the number of nerve processes increased in 10 and 100nmol/L groups, but there was no significant difference in cell survival rate and neurite number in 1nmol/L group (P 0.05). 1the length of nerve processes in 10 and 100nmol/L DHT groups was higher than that in model group (P < 0.05). After oxygen glucose deprivation / reoxygenation injury, the expression of RhoA mRNA and protein in the model group was significantly higher than that in the normal control group (P < 0.05), while the expression of Neuritin mRNA and protein in the model group was significantly lower than that in the normal control group (P < 0.05), DHT). 1the expression of RhoA mRNA and protein in 10 and 100nmol/L DHT groups was lower than that in model group (P < 0.05), while the expression of Neuritin mRNA and protein in 10 and RhoA mRNA group was higher than that in model group (P < 0.05). Conclusion: RhoA and Neuritin may be one of the important factors affecting neuronal regeneration in rat RGC-5 cells after oxygen-glucose deprivation / reoxygenation injury. DHT may play a neuroprotective role by regulating the expression of RhoA and Neuritin.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R774.6

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