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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 牟中原(Chung-Yuan Mou) | |
dc.contributor.author | Cheng-Shun Zheng | en |
dc.contributor.author | 鄭丞勛 | zh_TW |
dc.date.accessioned | 2021-06-15T16:35:02Z | - |
dc.date.available | 2020-08-16 | |
dc.date.copyright | 2015-08-16 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2015-08-12 | |
dc.identifier.citation | [1] Modi G, Pillay V, Choonara YE, 'Advances in the treatment of neurodegenerative disorders,' Ann. N.Y. Acad. Sci., vol. 1184, pp. 154-172, 2010.
[2] Ross CA, Poirier MA, 'Protein aggregation and neurodegenerative disease,' Nature Medicine, vol. 10, p. S10–S17, 2004. [3] Serrano-Pozo A, Frosch MP, Masliah E, Hyman BT., 'Neuropathological Alterations in Alzheimer Disease.,' Cold Spring Harb Perspect Med, vol. 1, 2011. [4] Rosales-Corral SA, Acuña-Castroviejo D, Coto-Montes A, Boga JA, Manchester LC, Fuentes-Broto L, Korkmaz A, Ma S, Tan DX, Reiter RJ., 'Alzheimer's disease: pathological mechanisms and the beneficial role of melatonin.,' Journal of Pineal Research, vol. 52, pp. 167-202, 2012. [5] Natalia Crespo-Biel, Clara Theunis, and Fred Van Leuven, 'Protein Tau: Prime Cause of Synaptic and Neuronal Degeneration in Alzheimer's Disease,' International Journal of Alzheimer's Disease, vol. 2012, 2012. [6] Alonso A, Zaidi T, Novak M, Grundke-Iqbal I, Iqbal K, 'Hyperphosphorylation induces self-assembly of tau into tangles of paired helical filaments/straight filaments,' Proc. Natl. Acad. Sci. U.S.A., vol. 98, pp. 6923-28, 2001. [7] Iqbal K, Alonso Adel C, Chen S, Chohan MO, El-Akkad E, Gong CX, Khatoon S, Li B, Liu F, Rahman A, Tanimukai H, Grundke-Iqbal I., 'Tau pathology in Alzheimer disease and other tauopathies,' Biochimica et Biophysica Acta (BBA), vol. 1739, pp. 198-210, 2005. [8] Mohandas E, Rajmohan V, Raghunath, 'Neurobiology of Alzheimer's disease.,' Indian J Psychiatry, vol. 51, pp. 55-61, 2009. [9] Hardy J, Selkoe DJ., 'The Amyloid Hypothesis of Alzheimer's Disease: Progress and Problems on the Road to Therapeutics,' Science, vol. 297, pp. 353-356, 2002. [10] Giri RK, Rajagopal V, Kalra VK., 'Curcumin, the active constituent of turmeric,inhibits amyloid peptide-induced cytochemokine gene expression and CCR5-mediated chemotaxis of THP-1 monocytes by modulating early growthresponse-1 transcription factor,' J. Neurochem., vol. 91, p. 1199–1210, 2004. [11] Aggarwal S, Ichikawa H, Takada Y, Sandur SK, Shishodia S, Aggarwal BB., 'Curcumin (diferuloylmethane) down-regulates expression of cell proliferation and antiapoptotic and metastatic gene products through suppression of IkappaBalpha kinase and Akt activation.,' Mol Pharmacol., vol. 69, pp. 195-206, 2006. [12] Kamat AM, Sethi G, Aggarwal BB., 'Curcumin potentiates the apoptotic effects of chemotherapeutic agents and cytokines through down-regulation of nuclear factor-κB and nuclear factor-κB-regulated gene products in IFN-α–sensitive and IFN-α–resistant human bladder cancer cells,' Mol Cancer Ther., vol. 6, pp. 1022-1030, 2007. [13] Kim DS, Park SY, Kim JK., 'Curcuminoids from Curcuma longa L. (Zingiberaceae) that protect PC12 rat pheochromocytoma and normal human umbilical vein endothelial cells from betaA(1-42) insult.,' Neurosci Lett, vol. 303, pp. 57-61, 2007. [14] Deng Y, Lu X, Wang L, Li T, Ding Y, Cao H, Zhang Y, Guo X, Yu G., 'Curcumin inhibits the AKT/NF-κB signaling via CpG demethylation of the promoter and restoration of NEP in the N2a cell line.,' AAPS J., vol. 16, pp. 649-657, 2014. [15] Lim GP, Chu T, Yang F, Beech W, Frautschy SA, Cole GM., 'The curry spice curcumin reduces oxidative damage and amyloid pathology in an Alzheimer transgenic mouse.,' The Journal of Neuroscience., vol. 21, pp. 8370-8377, 2001. [16] Anand P, Kunnumakkara AB, Newman RA, Aggarwal BB., 'Bioavailability of curcumin: problems and promises,' Mol. Pharm., vol. 4, pp. 807-818, 2007. [17] Yang KY, Lin LC, Tseng TY, Wang SC, Tsai TH., 'Oral bioavailability of curcumin in rat and the herbal analysis from Curcuma longa by LC-MS/MS.,' Journal of Chromatography B, vol. 853, pp. 183-189, 2007. [18] Hall A, 'Rho GTPases and the actin cytoskeleton.,' Science, vol. 279, no. 5350, pp. 509-514, 1998. [19] Sasaki T, Takai Y., 'The Rho small G protein family-Rho GDI system as a temporal and spatial determinant for cytoskeletal control.,' Biochem Biophys Res Commun., vol. 245, no. 3, pp. 641-645, 1998. [20] Negishi M, Katoh H., 'Rho Family GTPases as Key Regulators for Neuronal Network Formation.,' J. Biochem., vol. 132, pp. 157-166, 2002. [21] Katoh H, Yasui H, Yamaguchi Y, Aoki J, Fujita H, Mori K, and Negishi M., 'Small GTPase RhoG Is a Key Regulator for Neurite,' Mol. Cell. Biol., vol. 20, pp. 7378-7387, 2000. [22] Kresge CT,Leonowicz ME, Roth WJ, Vartuli JC, Beck JS, 'Ordered mesoporousmolecular sieves synthesized by a liquid-crystal template mechanism,' Nature, vol. 359, pp. 710-712, 1992. [23] Xia T, Kovochich M, Liong M, Meng H, Kabehie S, George S, Zink JI, Nel AE., 'Polyethyleneimine Coating Enhances the Cellular Uptake of Mesoporous Silica Nanoparticles and Allows Safe Delivery of siRNA and DNA Constructs,' ACS Nano, vol. 3, pp. 3273-3286, 2009. [24] Rosenholm JM, Peuhu E, Eriksson JE, Sahlgren C, Lindén M., 'Targeted Intracellular Delivery of Hydrophobic Agents using Mesoporous Hybrid Silica Nanoparticles as Carrier Systems.,' Nano Letters, vol. 9, pp. 3308-3311, 2009. [25] Slowing II, Trewyn BG, Lin VS, 'Mesoporous Silica Nanoparticles for Intracellular Delivery of Membrane-Impermeable Proteins,' JACS, vol. 129, pp. 8845-8849, 2007. [26] Liong M, Lu J, Kovochich M, Xia T, Ruehm SG, Nel AE, Tamanoi F, Zink JI., 'Multifunctional Inorganic Nanoparticles for Imaging, Targeting, and Drug Delivery.,' ACS Nano, vol. 2, pp. 889-896, 2008. [27] Mitra S, Gaur U, Ghosh PC, Maitra AN., 'Tumour targeted delivery of encapsulated dextran–doxorubicin conjugate using chitosan nanoparticles as carrier.,' J. Control. Release, vol. 74, pp. 317-323, 2001. [28] Wilson B, Samanta MK, Santhi K, Kumar KP, Ramasamy M, Suresh B., 'Chitosan nanoparticles as a new delivery system for the anti-Alzheimer drug tacrine.,' Nanomedicine, vol. 6, pp. 144-152, 2009. [29] Sonavane G, Tomoda K, Makino K., 'Biodistribution of colloidal gold nanoparticles after intravenous administration: effect of particle size.,' Colloids Surf B Biointerfaces., vol. 66, pp. 274-280, 2008. [30] Chen W, Tsai PH, Hung Y, Chiou SH, Mou CY., 'Nonviral Cell Labeling and Differentiation Agent for Induced Pluripotent Stem Cells Based on Mesoporous Silica Nanoparticles,' ACS Nano, vol. 7, p. 8423–8440, 2013. [31] Chen YP, Chen CT, Hung Y, Chou CM, Liu TP, Liang MR, Chen CT, Mou CY., 'A New Strategy for Intracellular Delivery of Enzyme Using,' J. Am. Chem. Soc., vol. 135, p. 1516−1523, 2013. [32] Suk JS, Suh J, Choy K, Lai SK, Fu J, Hanes J., 'Gene delivery to differentiated neurotypic cells with RGD and HIV Tat peptide functionalized polymeric nanoparticles.,' Biomaterials, vol. 27, pp. 5143-5150, 2006. [33] Liu L, Guo K, Lu J, Venkatraman SS, Luo D, Ng KC, Ling EA, Moochhala S, Yang YY., 'Biologically active core/shell nanoparticles self-assembled from cholesterol-terminated PEGeTAT for drug deliveryacross the bloodebrain barrier.,' Biomaterials, vol. 29, pp. 1509-1517, 2008. [34] Liang Y, Liu Z, Shuai X, Wang W, Liu J, Bi W, Wang C, Jing X, Liu Y, Tao E, 'Delivery of cationic polymer-siRNA nanoparticles for gene therapies in neural regeneration.,' Biochemical and Biophysical Research Communications, vol. 421, pp. 690-695, 2012. [35] Newland B, Dowd E, Pandit A., 'Biomaterial approaches to gene therapies for neurodegenerative disorders of the CNS.,' Biomater Sci, vol. 1, pp. 557-576, 2013. [36] Boussif O, Lezoualc'h F, Zanta MA, Mergny MD, Scherman D,Demeneix B, Behr JP., 'A versatile vector for gene and oligonucleotidetransfer into cells in culture and in vivo: polyethylenimine.,' ProcNatl Acad Sci USA, vol. 92, pp. 7297-7301, 1995. [37] Newland B, Moloney TC, Fontana G, Browne S, Abu-Rub MT, DowdE, Pandit AS., 'The neurotoxicity of gene vectors and its amelio-ration by packaging with collagen hollow spheres.,' Biomaterials, vol. 34, pp. 2130-2141., 2013. [38] Wilson B, Samanta MK, Santhi K, Kumar KP, Paramakrishnan N, Suresh B., 'Poly(n-butylcyanoacrylate) nanoparticles coated with polysorbate 80 for the targeted delivery of rivastigmine into the brain to treat Alzheimer's disease,' Brain Research, vol. 1200, pp. 159-168, 2008. [39] Tiwari SK, Agarwal S, Seth B, Yadav A, Nair S, Bhatnagar P, Karmakar M, Kumari M, Chauhan LK, Patel DK, Srivastava V, Singh D, Gupta SK, Tripathi A, Chaturvedi RK, Gupta KC., 'Curcumin-Loaded Nanoparticles Potently Induce Adult Neurogenesis and Reverse Cognitive Deficits in Alzheimer’s Disease Model via Canonical Wnt/β-Catenin Pathway,' ACS Nano, vol. 8, pp. 76-103, 2014. [40] Kim JA, Lee N, Kim BH, Rhee WJ, Yoon S, Hyeon T, Park TH., 'Enhancement of neurite outgrowth in PC12 cells by iron oxide nanoparticles.,' Biomaterials, vol. 32, pp. 2871-2877, 2011. [41] Madani F, Lindberg S, Langel U, Futaki S, Gräslund A., 'Mechanisms of cellular uptake of cell-penetrating peptides.,' J Biophys., vol. 2011, 2011. [42] Mishra S, Palanivelu K, 'The effect of curcumin (turmeric) on Alzheimer's disease: An overview,' Ann Indian Acad Neurol., vol. 11, pp. 13-19, 2008. [43] Negishi M, Katoh H., 'Rho Family GTPases as Key Regulators for Neuronal Network.,' J. Biochem, vol. 132, pp. 157-166, 2002. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/52939 | - |
dc.description.abstract | 阿茲海默症為一種神經退化性疾病且隨著年齡增長而增加,根據統計2014年全球超過4千萬人因病所苦。但目前為止,尚未找到有效的方法來治療或者延緩發病,因此治療阿茲海默症的方法,其研究及開發上相當的急迫性與重要性。
RhoG(質體DNA)屬於GTPases家族成員之一,根據過去文獻顯示RhoG藉由活化下游蛋白Rac與cdc42以調節細胞骨架誘導神經細胞分化。此外,薑黃素(Curcumin, CUR)萃取自薑黃,具有抗腫瘤以及抗發炎反應等藥物活性,過去在神經退化性疾病研究上發現其與發炎反應息息相關,因此薑黃素在治療阿茲海默症上非常具有潛力,然而薑黃素本身不溶於水,因此在生物應用上受到限制。 在本次研究上,我們使用PEG修飾的奈米材料(mesoporous silica nanoparticle,簡稱MSN)作為藥物載體,因為MSN具有中孔洞結構、易修飾和好的生物相容性等優點,能夠同時載入Curcumin (於中孔洞結構中)與攜帶RhoG(藉由材料表面修飾正電荷吸)。另外,為了增加細胞吞噬與基因轉染效率,我們引入TAT peptide於實驗設計上,我們稱此奈米藥物為CUR@FMSN(+)/RhoG-TAT。當CUR@FMSN(+)/RhoG-TAT被neuro-2a (N2a)細胞吞噬後,我們藉由流式細胞儀鑑定吞噬效率以及定量活性氧化物質(ROS),西方點墨法定量發炎反應相關蛋白表現,以及螢光顯微鏡觀察轉染結果。 在我們系統中,除了證明材料無生物毒性和好的細胞吞噬效率(接近99%吞噬)外並解決了薑黃素難溶於水所造成生物應用上的限制,此外我們也觀察到CUR@FMSN(+)/RhoG-TAT具有抗氧化、抗發炎及幫助神經生長等多重功能,因此我們認為MSN在神經退化性疾病的應用上具有相當大的潛力。 | zh_TW |
dc.description.abstract | Alzheimer's disease (AD), a progressive neurodegenerative disorder (ND) of the elderly, affects more than 44 million people worldwide. So far, there is no effective method that treats or delays the progression of AD. Therefore, development of strategies for AD therapy is very important goal.
RhoG, a member of small Rho family guanine nucleotide triphosphatases (GTPases), is able to induce neurite outgrowth through downregulating activation of Rac and cdc42, resulting in reorganizing the actin cytoskeleton and subsequent morphological changes. In addition, a well-known polyphenol compound, Curcumin, which was found from the rhizome of turmeric with antioxidant and anti- inflammatory properties, has already been noted for its therapy potential in neurodegenerative disease especially AD. However, due to limitations of poor bioavailability and aqueous solubility, curcumin is not suitable for biomedical applications. In our work, taking advantage of mesoporous silica nanoparticle (MSN), including its structure, easy modification and good biocompatibility, we proposed a concept of PEGylated MSN-based dual therapy and designed the nanodrug that contains curcumin(CUR) by MSN inner channel loading; MSN surface positive charge modification for RhoG gene adsorption; and TAT peptide enhanced the RhoG gene nuclear delivery and nonendocytosis mechanisms, named CUR@FMSN(+)/RhoG-TAT. MSN as a drug carrier improved the stability and bioavailability of curcumin. On the basis of the results shown in this study, Flow cytometry and confocal microscopy demonstrated the good cell uptake efficiency (approaching 99%) and localization of CUR@FMSN(+)/RhoG-TAT in neuro-2a (N2a) cell line. The expression levels of inflammatory related proteins inhibited by CUR@FMSN(+)/RhoG-TAT were carried out by using Western blotting. The levels of paraquat-induced ROS reduced by nanodrug were stained by DHE assays and quantified by flow cytometry. Our results evaluated that CUR@FMSN(+)/RhoG-TAT not only stimulated neurite outgrowth but also allowed neuron cells to against ROS-induced damage in N2a cells. Hence, the application of MSN might be a promising candidate method for treating NDs. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T16:35:02Z (GMT). No. of bitstreams: 1 ntu-104-R02223124-1.pdf: 3645706 bytes, checksum: c8c69d894c9c8c061bcf8aa28e103e55 (MD5) Previous issue date: 2015 | en |
dc.description.tableofcontents | 謝誌 I
中文摘要 II Abstract IV Table of Content VI List of Figure VIII List of Schemes XII List of Table XIII Chapter 1 Introduction..................................1 1.1 Neurodegenerative Diseases..........................1 1.1.1 Tau Hypothesis..................................2 1.1.2 Amyloid Hypothesis..............................4 1.2 Role of Curcumin in Health and Disease..............5 1.3 Role of Rho Family Small GTPases....................8 1.4 Mesoporous Silica Nanoparticles (MSN)..............10 1.4.1 Brief History of MSN...........................10 1.4.2 Multifunctional Mesoporous Silica Nanoparticles .......................................................11 1.5 Desirable structural and physicochemical properties of nanoparticles (NPs).................................12 1.6 Literature Review of Nanoparticles Applications In Neurodegenerative Diseases (ND)........................17 1.6.1 Gene delivery..................................17 1.6.2 Drug delivery..................................19 1.7 Summary............................................20 Chapter 2 Experimental Section.........................22 2.1 Materials..........................................22 2.2 Synthesis of Green Fluorescent Mesoporous Silica Nanoparticles (FMSN(+))................................23 2.3 Dynamic Light Scattering (DLS).....................24 2.4 Transmission Election Microscopy (TEM).............24 2.5 N2 Adsorption-desorption Isotherms.................25 2.6 X-ray Diffraction (XRD) Patterns...................25 2.7 Agarose Gel Electrophoresis........................25 2.8 Cell Line and Cell Culture.........................26 2.9 Cytotoxicity of FMSN(+)............................26 2.10 Flow Cytometry Analysis...........................27 2.11 Loading efficiency of CUR in FMSN(+)..............28 2.11.1 Synthesis of CUR@FMSN(+).......................28 2.11.2 Examine Loading Efficiency of CUR in FMSN(+) .......................................................28 2.12 ROS measurement with DHE by Flow Cytometry........29 2.13 Measurement of Neurite Length of N2a cells........30 2.14 Western Blot Analysis.............................32 2.15 Confocal Microscopy...............................33 Chapter 3 Results and Discussion.......................34 3.1 Characterization and Surface Functionalization of FMSN(+)................................................34 3.1.1 Dynamic Light Scattering (DLS) and Zeta Potential Measurements...........................................36 3.1.2 N2 Adsorption-desorption Isotherm Measurements .......................................................37 3.2 Cytotoxicity and Cellular Uptake Efficiency of FMSN(+) Nanoparticles..................................38 3.3 Plasmid RhoG binding abilities of TA-modified FMSN (FMSN(+))..............................................42 3.4 Effect of TAT Peptide on the Transfection and Delivery of FMSN(+)/RhoG-TAT...........................43 3.5 Flow Cytometry Examinations of the Effect of Endocytosis Inhibitors on the Cellular uptake of FMSN(+)s...............................................47 3.6 Loading efficiency and Loading amount of CUR in FMSN(+)................................................49 3.7 Protective Effect of CUR on PQ Oxidative Stress .......................................................51 3.8 Quantification of Neurite Outgrowth Protected by CUR@MSN(+)/RhoG-TAT treatment..........................54 3.9 CUR@FMSN(+)/RhoG-TAT inhibits the Expression of Inflammatory-related proteins in N2a cells.............56 Chapter 4 Conclusion...................................59 | |
dc.language.iso | en | |
dc.title | 雙功能中孔洞奈米藥物載體於治療神經退化性疾病之設計 | zh_TW |
dc.title | Design of Nanodrug for Neurodegenerative Disease by using Dual-functionalized Mesoporous Silica Nanoparticle | en |
dc.type | Thesis | |
dc.date.schoolyear | 103-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 簡汎清(Fan-Ching Chien),戴桓青(Hwan-Ching Tai) | |
dc.subject.keyword | 中孔洞二氧化矽奈米粒子,神經退化性疾病,阿茲海默症,薑黃素,藥物傳遞系統,基因治療, | zh_TW |
dc.subject.keyword | Mesoporous silica nanoparticles,Neurodegenerative diseases,Curcumin,Rho family GTPases,Drug delivery,Gene delivery, | en |
dc.relation.page | 63 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2015-08-12 | |
dc.contributor.author-college | 理學院 | zh_TW |
dc.contributor.author-dept | 化學研究所 | zh_TW |
顯示於系所單位: | 化學系 |
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