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研究生(外文):Hsuan-Chi Huang
論文名稱(外文):Inhibition of nitric oxide production promotes microtubule assembly in the reconnected facial nerve
指導教授(外文):Pei-Hsin Liu
外文關鍵詞:facial nerveaxotomynerve conduction studymicrotubulenitric oxide
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我們最近報導,顏面神經截斷再縫合後,於腹膜腔內注射一氧化氮合成�〞漣磻蹌砲i促進受傷顏面運動神經元軸突的再生。其機制可能與神經元細胞體內的自由基環境有關,因為給予一氧化氮合成�〞漣磻蹌祕P時也抑制了細胞內一氧化氮合成�〞漯穛{大幅增加。然而,自由基對於軸突再生所產生的效應未必要間接透過影響神經元細胞體,而可直接作用在神經元軸突。最近的研究顯示,一氧化氮及peroxynitrite可對神經元的微小管蛋白及微小管相關蛋白造成氧化傷害。因此,本研究藉由in vitro及in vivo的方法探討顏面神經截斷再縫合後,一氧化氮合成�“磻蹌笆鴭鬊C面神經內細胞骨架微小管所產生的影響。電生理檢測結果顯示,將一氧化氮合成�“磻蹌烊-NAME注射到再接合的顏面神經中,為時二週,比一氧化氮生成劑GSNO更能促進軸突再生及傳導功能回復。此變化可能與軸突內的微小管有關,因為抑制一氧化氮合成同時也提高微小管蛋白的聚合活性及tau (神經元特有的微小管相關蛋白) 幫助微小管聚合的能力。相反地,無論是in vitro或in vivo給予GSNO皆抑制微小管蛋白的聚合。我們也發現顏面神經縫合處的近端神經段中,若給予GSNO會比L-NAME造成α-tubulin及tau 有較大程度的tyrosine nitration及phosphorylation,以及減低α-tubulin的acetylation。我們的研究結果顯示,抑制一氧化氮的產生可促進微小管聚合,也可能因此有助於顏面神經元軸突的再生。微小管聚合的活性可能部分藉由微小管蛋白及tau的修飾來調控。直接於再接合周邊神經給予一氧化氮合成�“磻蹌砲i能可作為周邊神經修復的輔助治療方式。
We recently reported that intraperitoneal injection of nitric oxide synthase (NOS) inhibitors promotes facial axonal regeneration following neurorrhaphy. The underlying mechanism may involve the intracellular free radical environment since treatment of NOS inhibitors prevents the intraneuronal large-scale upregulation of NOS. However, the free radical-mediated effect on axonal regeneration may not need to occur indirectly via influencing on the neuronal cell body but instead directly on the neuronal axon. Here we investigated the effect of NOS inhibitor on the microtubule cytoskeleton within the repaired facial axons via combined in vitro and in vivo approaches since neuronal tubulins, the building blocks of microtubules, and microtubule-associated proteins (MAPs) have been demonstrated as candidate targets to NO/ONOO−. Electrophysiology revealed that application of a NOS inhibitor N-nitro-L-arginine methyl ester (L-NAME), rather than a NO donor S-nitrosoglutathione (GSNO), into reconnected facial nerves for 2 weeks facilitates axonal regeneration and functional recovery. These changes may be associated with axonal microtubules since NOS inhibition enhanced the activity of tubulin polymerization and the ability of tau, a neuron-specific MAP, to promote microtubule assembly. On the contrary, both in vitro and in vivo treatment with GSNO, prohibited tubulin polymerization. We also show that application of GSNO, rather than L-NAME, resulted in larger extent of tyrosine nitration and phosphorylation of α-tubulin and tau, as well as less extent of acetylation of α-tubulin in the proximal stump of reconnected facial nerve. Our studies suggest that inhibition of NO production promotes microtubule assembly and may thus assist in facial axonal regeneration. The activity of microtubule polymerization may be in part mediated by modifications of tubulins and tau. Application of NOS inhibitor into reconnected peripheral nerve may be an assistant therapeutic strategy to peripheral nerve repair.
英文摘要 …………………………… 1
中文摘要 …………………………… 2
引言 ………………………………… 3
材料與方法 ………………………… 7
結果 ………………………………… 11
討論 ………………………………… 16
結論 ………………………………… 22
參考文獻 …………………………… 23
圖表與圖片說明 …………………… 29
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