以細胞培養及動物模式探討蕈毒鹼受體拮抗劑阿托品對神經纖維生長及細胞存活之影響

Effect of muscarinic receptor antagonist atropine on neurite outgrowth and neuronal survival: An in vitro and in vivo study

賴孟琳(Meng-Lin Lai)

指導教授 ： 曾廣文 ； 陳賢堂

中山醫學大學/醫學科技學院/生物醫學科學學系碩士班/碩士(2012年)

https://doi.org/10.6834/CSMU.2012.00080

摘要

近年來，台灣近視比例有持續攀升的趨勢，然而對於近視形成的機制仍尚未釐清。阿托品（atropine）是一種非選擇性蕈毒鹼拮抗劑（non-selective muscarinic receptor antagonist），臨床上用來做為近視治療方針已有一段時間。然而，對於阿托品是否會對眼睛組織造成影響仍是未知。為了證明阿托品對於神經纖維生長及細胞存活的影響，本研究利用實驗用小白鼠及存有蕈毒鹼受體的神經瘤母細胞進行探討阿托品對於眼睛所造成之影響。動物實驗中，我們每天以1 %阿托品處理小鼠連續8至32週，並利用形態學分析證實，阿托品會導致小鼠虹膜神經纖維比例以及睫狀神經節中神經元細胞數量減少。體外細胞培養實驗中，和對照組相比之下，經由加入0.15、0.75以及1.5 μM阿托品第一、二和三天後，會導致細胞形態縮小，並且出現類似凋亡細胞核濃染的情形。另外，在加入阿托品後同時也會導致細胞之神經纖維縮短。而加入0.15、0.75以及1.5 μM阿托品處理一、二和三天後，免疫染色結果發現，細胞凋亡標記蛋白caspase-3會隨著阿托品濃度增加及點藥天數的增長，表現量有上升的趨勢，證實加入阿托品的確會使細胞走向凋亡之路徑。本研究的結果表明了，阿托品會對小鼠的虹膜神經纖維以及睫狀神經節造成影響，並且會使得細胞走向凋亡。
關鍵字: 阿托品、虹膜、蕈毒鹼受體、凋亡

關鍵字

阿托品 ； 虹膜 ； 蕈毒鹼受體 ； 凋亡

並列摘要

The prevalence of myopia has become increasingly high in recent years in Taiwan. At present, the exact mechanism of myopia remains unclear. Atropine, a pan muscarinic cholinergic receptor antagonist, has been used for treated myopia since long time ago. However, the effect of atropine on ocular tissue is not known. In order to investigate the influence of atropine in neurite outgrowth and cell death, atropine-treatment was examined in BALB/cByJNarl mice and mouse neuroblastoma cells. In vivo, mice were treated with 1% atropine every day ranging 8 to 32 weeks. Morphological results showed that the number of neurons within the ciliary ganglion and the autonomic innervation density of the iris were reduced after treatment with atropine. In vitro, neuroblastoma cells were cultured and treated with atropine for 1, 2 and 3 days. After the cell treated with atropine, the morphology of neuron2a cells was smaller than control, and the apoptosis-like characteristic of nuclei condensation could be observed. In addition, atropine caused the cells neurite lengths decreased compared with control. Atropine induced time- and dose-dependent caspase-3 proteins activity was detected in neuron2a cells. These results indicate that atropine invading induced nerve innervation and cell death.

Keyword: atropine, iris, muscarinic receptor, apoptosis

並列關鍵字

atropine ； iris ； muscarinic receptor ； apoptosis

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