Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/20196
標題: 利用肌肉去神經化模式探討肌肉注射之羊水幹細胞增進損傷神經之再生程度
Increased Nerve Regeneration by Intramuscular Injection of Human Amniotic Fluid Mesenchymal Stem Cells in a Muscle Denervation Model
作者: 呂宗翰
Lu, Zong-Han
關鍵字: 神經滋養因子;neurotrophic factors;神經再生;肌肉去神經化;羊水幹細胞;nerve regeneration;muscle denervation;amniotic fluid mesenchymal cells
出版社: 生物醫學研究所
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摘要: 
Abstract
Purpose: Neurotrophic factors provide the basis for neurotrophic signaling within muscle compartments essential for muscle regeneration in muscle denervation. It is well known that human amniotic fluid mesenchymal cells (AFS) have the potential to secrete various neurotrophic factors mandatory for nerve regeneration. In this study, we assess the outcome of nerve regeneration by intramuscular injection of AFS in a muscle denervation and nerve anastomosis model.
Materials and methods: Sprague-Dawley rats weighting 200-250 gm were enrolled in this study. Muscle denervation was conducted by transverse resection of sciatic nerve with proximal end sutured into the gluteal muscle. The nerve anastomosis model was performed by transverse resection of sciatic nerve followed by 4 stitches suture. AFS with 5x106 cells were intramuscularly injected to gastrocnemius muscle with infusion pump.
Results: CNTF, BDNF, NT-3 and GDNF were remarkably expressed in AFS cells. Intra-muscular injection of AFS exerted significantly expression of several neurotrophic factors over nerve and innervated muscle. AFS caused high expression of Bcl-2 in denervated muscle with reciprocal decrease of Bad and Bax. AFS preserved the muscle morphology paralleling with high expression of desmin and acetylocholine receptors. AFS injection produced the significant improvement in neurobehavior such as CatWalk gait analysis as well as nerve conduction latency and CMAP. Significant perseveration of anterior horn cell and increased nerve myelination was line with muscle morphology.
Conclusion: Intramuscular injection of AFS protects muscle apoptosis by the secretion of various neurotrophic factors. This
protection furthermore improves the nerve regeneration in long
term nerve anastmosis model.

中文摘要
目的:神經營養因子是失去神經支配的肌肉,再生時所不可或缺的重要因素。目前研究指出,人類羊水幹細胞(AFS)具有分泌多種神經營養因子的潛力,能夠協助神經的再生。在這項研究中,我們將羊水幹細胞,注射到完全去神經化及神經縫合,這兩種動物模式的肌肉中,藉此來評估神經肌肉的再生。
材料與方法:我們使用體重200~250克的SD大鼠進行這項研究。在完全去神經化的動物模式中,將坐骨神經進行橫向切除,並將其近心端縫到臀部肌肉上。神經縫合的動物模型,進行橫向切除坐骨神經後將兩端縫合4針。用微量注射器將5×106羊水幹細胞注射到腓腸肌中。
結果:睫狀神經營養因子(CNTF)、腦源性神經滋養因子(BDNF)、神經滋養因子3(NT-3)和神經膠質細胞神經營養因子(GDNF)在羊水幹細胞中顯著的表達。肌肉內注射的羊水幹細胞,顯著表達一些神經營養因子,在神經和受神經支配的肌肉中。羊水幹細胞造成失去神經支配的肌肉,增加B-細胞淋巴瘤基因2(Bcl-2)的表現量,並減少Bcl-2相關死亡促進蛋白(Bad)和Bcl-2相關X蛋白(Bax)的表現量。羊水幹細胞能協助維持肌肉的形態,與之相匹配的標的是結蛋白(Desmin)及乙醯膽鹼受體(acetylcholine receptor)的高表現量。羊水幹細胞注射後,神經功能得到顯著改善如CatWalk步態分析,以及神經傳導速率(Latency) 和複合肌肉動作電位波(compound muscle action potential)。有效的改善前角細胞和增加神經的髓鞘及下游的肌肉形態。
結論:羊水幹細胞注射於肌肉後,分泌多種神經營養因子能夠防止肌肉細胞凋亡。這種保護在長期的神經縫合模式中,可以進一步提高神經與肌肉的再生。
URI: http://hdl.handle.net/11455/20196
其他識別: U0005-0307201306001200
Appears in Collections:生物醫學研究所

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