碩士 / 國立臺灣師範大學 / 生命科學研究所 / 101 / Alzheimer's disease (AD) is a progressive neurodegenerative disorder associated with accumulation of extracellular amyloid-β and intracellular tau-associated neurofibrillary tangles (NFTs). The aggregation of tau in AD correlates with the clinical progression of the disease and inhibition
or reversal of tau aggregation may protect the affected neurons. In this study wild type (K18) and pro-aggregation mutant (ΔK280) of repeat domain of tau (tauRD) was fused with DsRed (tauRD-DsRed fusion gene) and used to generate Tet-On 293 and SH-SY5Y cell clones as drug screening platforms. Inhibitors that retard or block ΔK280 tau aggregation can be distinguished by increasing fluorescence on Tet-On 293 cells. For those compounds/herbs with putative aggregate inhibitors, mechanisms of neuroprotection were determined. Upon induction with
doxycycline, red fluorescence in ΔK280 tauRD-DsRed 293 cells was significantly reduced compared to that in wild type cells. As a positive control, congo red increased red fluorescence more effectively in ΔK280 tauRD-DsRed 293 cells than in wild type cells. Without significantly
reducing cell numbers, pretreatment of NC009-1, -2, -3, -6, -7 (indolylquinoline compounds), NTNU-003, -008 (GSK-3 inhibitor-like compounds), NH021, NTNU-043, -057, -059, -224, -309, -313, -319, -331, -379 (herbal extracts) resulted in significant increased fluorescence on ΔK280 tauRD-DsRed cells. For SH-SY5Y cell clones, retinoic acid
treatment generated cells resemblance to adult neurons. Fluorescent microscopy examination revealed that neurite outgrowth (including total outgrowth, process and branch) was significantly reduced in ΔK280 tauRD-DsRed cells compared to that in wild type cells. NC009-1, -7 and
NTNU-008 significantly increased neurite total outgrowth in ΔK280 SH-SY5Y cells, accompanying with enhanced HSP27 (NC009-1 and -7) or GRP78 and HSP27 (NTNU-008) expression.
| Identifer | oai:union.ndltd.org:TW/101NTNU5112049 |
| Date | January 2013 |
| Creators | Hsuan-Chiang Chen, 陳炫江 |
| Contributors | Guey-Jen Lee-Chen, 李桂楨 博士 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 53 |
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