跳到主要內容

臺灣博碩士論文加值系統

(34.226.244.254) 您好!臺灣時間:2021/08/03 04:13
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

: 
twitterline
研究生:王婉蓉
研究生(外文):Wang, Wanerong
論文名稱:角膜緣纖維母細胞的神經保護作用
論文名稱(外文):Protective Effect of Limbal Fibroblasts Against Neurotoxicity
指導教授:馬國興馬國興引用關係梁章敏黃庸協
指導教授(外文):Ma, KuohsingLiang, ChangminHuang, Yuanshieh
口試委員:馬國興黃庸協劉江川
口試委員(外文):Ma, KuohsingHuang, YuanshiehLiu, Jiangchuan
口試日期:2012-05-24
學位類別:碩士
校院名稱:國防醫學院
系所名稱:生物及解剖學研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:51
中文關鍵詞:人類角膜緣纖維母細胞神經保護中腦細胞6-羥多巴胺鹽酸鹽
外文關鍵詞:Human limbal fibroblastsNeuroprotectVentral mesencephalic cells6-hydroxydopamine hydrochloride
相關次數:
  • 被引用被引用:0
  • 點閱點閱:138
  • 評分評分:
  • 下載下載:3
  • 收藏至我的研究室書目清單書目收藏:0
眼睛角膜緣基質中的細胞群體稱為角膜緣纖維母細胞,除了提供養分給周遭上皮細胞以維持一般的生理機制外,相較於兩側的角膜與鞏膜基質區,更擁有獨特的功能幫助角膜緣上皮幹細胞進行自我更新及分化。近幾年發現,這些細胞能夠簡易的分離出來進行體外培養與增生,經過處理更可以分化成不同譜系的細胞,甚至有非常類似於骨髓間葉幹細胞能夠分泌神經滋養因子的特性。由於近年來間葉幹細胞在神經退化性疾病的治療研究上十分具有潛力,除了處理分化成神經元以外,更由於其能分泌神經滋養或生長因子,而能夠幫助受損腦區的神經元或纖維進行修復與再生。因此我們的假設是,角膜緣纖維母細胞是否能夠藉由其分泌的物質以保護神經元不受到神經毒性傷害。本研究利用巴金森氏症的細胞模式,培養富含多巴胺神經元的中腦細胞,以6-OHDA的神經毒性造成細胞傷害,並收集角膜緣纖維母細胞的條件培養液來共同處理。實驗結果發現,曾暴露於6-OHDA神經毒性的多巴胺神經元經過條件培養液處理,其存活率不僅能夠提升,同時避免其神經纖維及分支數量的萎縮,且在神經元突觸互相連接的部位也可以觀察多分支的情形。此外,非多巴胺神經元的神經細胞也能受到條件培養基保護,提升整體細胞之存活率。本實驗證實,初代培養的角膜緣纖維母細胞可分泌具有神經保護以及促進神經纖維生長的因子,如VEGF、IGF-II、TGF-β和HGF,可做為研究治療神經退化性疾病的一種細胞材料。
Limbal fibroblasts in the stroma layer of the limbus have BM-MSC-like properties. It has been reported that LFs express genes of NT-3, GDNF, GDNF precursor and BDNF. The neurotrophic factors can maintain neurons survive and protect neurons against the oxidative injury. Parkinson's disease (PD) is caused by progressive degeneration of dopaminergic neurons in the substantia nigra pars compacta. The mechanism of dopaminergic neuron cell death in Parkinson’s disease (PD) is finally encountering the oxidative stress-induced effect, and it could be rescued by neurotrophic factors. Our hypothesis is that limbal fibroblast might be able to secret some neurontrophic factors to prevent neuronal damages and support survival of dopaminergic neurons enriched ventral messencephalic cells. In our studies, at first, we evaluate the ventral messencephalic cells survival rate with 6-OHDA in different does and time. Second, we use the ventral messencephalic cells treated with 6-hydroxydopamine hydrochloride (6-OHDA) for1 hour and then transferred to a new culture environment with or without fibroblast conditional medium. After all, we evaluated the survival rate of dopaminergic neurons by immunochemical stain, and the secretory growth factors in conditional medium were analyzed with RayBio® Human growth factor antibody array. We found the survival rate of ventral messencephalic cells treated with conditional medium is increased significantly. In the result of antibody array, there are some factors associated with neuronprotection, such as VEGF, TGF-b IGF-2 and HGF, in the conditional medium. In our study, we suggested that limbal fibroblasts could protect ventral messencephalic cells from cell death by 6-OHDA induced oxidative stress. The cocktail of secretory growth factors may contribute to the protective effects.
目錄……………………………………………………………I
圖目錄…………………………………………………………III
中文摘要………………………………………………………IV
英文摘要………………………………………………………VI
第一章、緒論…………………………………………………1
第一節、角膜緣纖維母細胞…………………………………1
第二節、神經營養因子………………………………………2
第三節、多巴胺神經元………………………………………3
第四節、巴金森氏症…………………………………………5
第五節、間葉幹細胞治療神經退化性疾病…………………8
第六節、巴金森氏症細胞模式………………………………9
第七節、實驗目的……………………………………………10
第二章、材料與方法…………………………………………12
第一節、胚胎大鼠中腦細胞…………………………………12
第二節、Poly-D-lysine鍍膜蓋玻片……………………13
第三節、人類角膜緣纖維母細胞……………………………13
第四節、條件培養液…………………………………………13
第五節、6-OHDA毒性反應…………………………………14
第六節、條件培養液對6-OHDA處理的中腦多巴胺神經元影響14
第七節、免疫螢光染色………………………………………15
第八節、免疫細胞螢光染色結果定量分析……………………16
第九節、蛋白質陣列系統(Protein array)…………………16
第十節、酵素免疫分析法(Enzyme-linked immunoassay)16
第十一節、神經纖維定量分析…………………………………16
第三章、結果……………………………………………………17
第一節、初代胚胎中腦細胞培養………………………………17
第二節、6-OHDA毒性影響中腦細胞及多巴胺神經元存活率…18
第三節、角膜緣纖維母細胞對神經細胞的保護作用……………20
第四節、條件培養液蛋白陣列分析………………………………21
第五節、角膜緣纖維母細胞分泌血管新生素VEGF………………22
第四章、討論………………………………………………………23
第五章、結論………………………………………………………27
第六章、參考文獻…………………………………………………29

Amirjamshidi, H., B. Y. Milani, et al. (2011). "Limbal fibroblast conditioned media: a non-invasive treatment for limbal stem cell deficiency." Mol Vis 17: 658-666.
Balasooriya, I. S. W., Kandatege (2007
). "Are SH-SY5Y and MN9D cell lines truly dopaminergic?" Proceedings: 3rd Annual Symposium: Graduate Research and Scholarly Projects.
Bayer, S. A., K. V. Wills, et al. (1995). "Time of neuron origin and gradients of neurogenesis in midbrain dopaminergic neurons in the mouse." Exp Brain Res 105(2): 191-199.
Beck, K. D., J. Valverde, et al. (1995). "Mesencephalic dopaminergic neurons protected by GDNF from axotomy-induced degeneration in the adult brain." Nature 373(6512): 339-341.
Benchenane, K., J. P. Lopez-Atalaya, et al. (2004). "Equivocal roles of tissue-type plasminogen activator in stroke-induced injury." Trends Neurosci 27(3): 155-160.
Biedler, J. L., S. Roffler-Tarlov, et al. (1978). "Multiple neurotransmitter synthesis by human neuroblastoma cell lines and clones." Cancer Res 38(11 Pt 1): 3751-3757.
Bjorklund, T. and J. H. Kordower (2010). "Gene therapy for Parkinson's disease." Mov Disord 25 Suppl 1: S161-173.
Blandini, F. (2009). "Neuroprotective compounds and innovative
therapeutic strategies for Parkinson’s disease:
experimental and clinical studies." Open Access Journal of Clinical Trials 1 1–15.
Blandini, F., L. Cova, et al. (2010). "Transplantation of undifferentiated human mesenchymal stem cells protects against 6-hydroxydopamine neurotoxicity in the rat." Cell Transplant 19(2): 203-217.
Blum, D., S. Torch, et al. (2001). "Molecular pathways involved in the neurotoxicity of 6-OHDA, dopamine and MPTP: contribution to the apoptotic theory in Parkinson's disease." Prog Neurobiol 65(2): 135-172.
Chen, R. C., S. F. Chang, et al. (2001). "Prevalence, incidence, and mortality of PD: a door-to-door survey in Ilan county, Taiwan." Neurology 57(9): 1679-1686.
Chen, W. F., C. Chakraborty, et al. (2012). "Neuroprotection by marine-derived compound, 11-dehydrosinulariolide, in an in vitro Parkinson's model: a promising candidate for the treatment of Parkinson's disease." Naunyn Schmiedebergs Arch Pharmacol 385(3): 265-275.
Cheng, B. and M. P. Mattson (1991). "NGF and bFGF protect rat hippocampal and human cortical neurons against hypoglycemic damage by stabilizing calcium homeostasis." Neuron 7(6): 1031-1041.
Chinta, S. J. and J. K. Andersen (2005). "Dopaminergic neurons." Int J Biochem Cell Biol 37(5): 942-946.
Choi, H. K., L. Won, et al. (1992). "Specific modulation of dopamine expression in neuronal hybrid cells by primary cells from different brain regions." Proc Natl Acad Sci U S A 89(19): 8943-8947.
Cohen, S., R. Levi-Montalcini, et al. (1954). "A NERVE GROWTH-STIMULATING FACTOR ISOLATED FROM SARCOM AS 37 AND 180." Proc Natl Acad Sci U S A 40(10): 1014-1018.
Cova, L., M. T. Armentero, et al. (2010). "Multiple neurogenic and neurorescue effects of human mesenchymal stem cell after transplantation in an experimental model of Parkinson's disease." Brain Res 1311: 12-27.
Crigler, L., R. C. Robey, et al. (2006). "Human mesenchymal stem cell subpopulations express a variety of neuro-regulatory molecules and promote neuronal cell survival and neuritogenesis." Exp Neurol 198(1): 54-64.
Dawson, T. M. and V. L. Dawson (2003). "Rare genetic mutations shed light on the pathogenesis of Parkinson disease." J Clin Invest 111(2): 145-151.
Deacon, T., J. Schumacher, et al. (1997). "Histological evidence of fetal pig neural cell survival after transplantation into a patient with Parkinson's disease." Nat Med 3(3): 350-353.
Dravida, S., R. Pal, et al. (2005). "The transdifferentiation potential of limbal fibroblast-like cells." Brain Res Dev Brain Res 160(2): 239-251.
Dunn, E. H. (1917). "Primary and secondary findings in a series of attempts to transplant cerebral cortex in the albino rat." J Comp Neurol 27: 565–582.
Freed, C. R., P. E. Greene, et al. (2001). "Transplantation of embryonic dopamine neurons for severe Parkinson's disease." N Engl J Med 344(10): 710-719.
Ganser, C., A. Papazoglou, et al. (2010). "Neuroprotective effects of erythropoietin on 6-hydroxydopamine-treated ventral mesencephalic dopamine-rich cultures." Exp Cell Res 316(5): 737-746.
Gasser, T. (2001). "Genetics of Parkinson's disease." J Neurol 248(10): 833-840.
Graham, D. G., S. M. Tiffany, et al. (1978). "Autoxidation versus covalent binding of quinones as the mechanism of toxicity of dopamine, 6-hydroxydopamine, and related compounds toward C1300 neuroblastoma cells in vitro." Mol Pharmacol 14(4): 644-653.
Halliwell, B. (1992). "Reactive oxygen species and the central nervous system." J Neurochem 59(5): 1609-1623.
Han, B. S., H. S. Hong, et al. (2003). "Caspase-dependent and -independent cell death pathways in primary cultures of mesencephalic dopaminergic neurons after neurotoxin treatment." J Neurosci 23(12): 5069-5078.
Hornykiewicz, O. (1975). "Parkinson's disease and its chemotherapy." Biochem Pharmacol 24(10): 1061-1065.
Jeon, B. S., V. Jackson-Lewis, et al. (1995). "6-Hydroxydopamine lesion of the rat substantia nigra: time course and morphology of cell death." Neurodegeneration 4(2): 131-137.
Joyce, N., G. Annett, et al. (2010). "Mesenchymal stem cells for the treatment of neurodegenerative disease." Regen Med 5(6): 933-946.
Karlsson, J., M. Emgard, et al. (2002). "Comparison between survival of lazaroid-treated embryonic nigral neurons in cell suspensions, cultures and transplants." Brain Res 955(1-2): 268-280.
Kingwell, K. (2011). "Parkinson disease: Gene therapy for Parkinson disease shows promise in phase II trial." Nat Rev Neurol 7(5): 241.
Koike, H., A. Ishida, et al. (2006). "Prevention of onset of Parkinson's disease by in vivo gene transfer of human hepatocyte growth factor in rodent model: a model of gene therapy for Parkinson's disease." Gene Ther 13(23): 1639-1644.
Langston, J. W., E. B. Langston, et al. (1984). "MPTP-induced parkinsonism in human and non-human primates--clinical and experimental aspects." Acta Neurol Scand Suppl 100: 49-54.
Li, W., Y. Hayashida, et al. (2007). "Niche regulation of corneal epithelial stem cells at the limbus." Cell Res 17(1): 26-36.
Liu, B. and J. S. Hong (2003). "Primary rat mesencephalic neuron-glia, neuron-enriched, microglia-enriched, and astroglia-enriched cultures." Methods Mol Med 79: 387-395.
Mackay, K. B., S. A. Loddick, et al. (2003). "Neuroprotective effects of insulin-like growth factor-binding protein ligand inhibitors in vitro and in vivo." J Cereb Blood Flow Metab 23(10): 1160-1167.
Marti, J., K. V. Wills, et al. (2002). "A combined immunohistochemical and autoradiographic method to detect midbrain dopaminergic neurons and determine their time of origin." Brain Res Brain Res Protoc 9(3): 197-205.
Mattson, M. P. (2008). "Glutamate and neurotrophic factors in neuronal plasticity and disease." Ann N Y Acad Sci 1144: 97-112.
Muller, L. J., C. F. Marfurt, et al. (2003). "Corneal nerves: structure, contents and function." Exp Eye Res 76(5): 521-542.
Peterson, A. L. and J. G. Nutt (2008). "Treatment of Parkinson's disease with trophic factors." Neurotherapeutics 5(2): 270-280.
Polisetti, N., P. Agarwal, et al. (2010). "Gene expression profile of epithelial cells and mesenchymal cells derived from limbal explant culture." Mol Vis 16: 1227-1240.
Polisetty, N., A. Fatima, et al. (2008). "Mesenchymal cells from limbal stroma of human eye." Mol Vis 14: 431-442.
Prakash, N. and W. Wurst (2006). "Development of dopaminergic neurons in the mammalian brain." Cell Mol Life Sci 63(2): 187-206.
Seeberger, L. C. and R. A. Hauser (2009). "Levodopa/carbidopa/entacapone in Parkinson's disease." Expert Rev Neurother 9(7): 929-940.
Simon, H. H., S. Thuret, et al. (2004). "Midbrain dopaminergic neurons: control of their cell fate by the engrailed transcription factors." Cell Tissue Res 318(1): 53-61.
Smith, P. F. (2008). "Inflammation in Parkinson's disease: an update." Curr Opin Investig Drugs 9(5): 478-484.
Takeuchi, Y., T. Sawada, et al. (1991). "Effects of 6-hydroxydopamine lesions of the nigrostriatal pathway on striatal serotonin innervation in adult rats." Brain Res 562(2): 301-305.
Wakeman, D. R., H. B. Dodiya, et al. (2011). "Cell transplantation and gene therapy in Parkinson's disease." Mt Sinai J Med 78(1): 126-158.
Wilkins, A., K. Kemp, et al. (2009). "Human bone marrow-derived mesenchymal stem cells secrete brain-derived neurotrophic factor which promotes neuronal survival in vitro." Stem Cell Res 3(1): 63-70.
Yasuhara, T., T. Shingo, et al. (2007). "Glial cell line-derived neurotrophic factor (GDNF) therapy for Parkinson's disease." Acta Med Okayama 61(2): 51-56.
Zawada, W. M., D. L. Kirschman, et al. (1996). "Growth factors rescue embryonic dopamine neurons from programmed cell death." Exp Neurol 140(1): 60-67.

QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top