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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 黃銓珍(Chang-Jen Huang) | |
| dc.contributor.author | Yi- Lin Huang | en |
| dc.contributor.author | 黃意霖 | zh_TW |
| dc.date.accessioned | 2021-06-16T02:56:52Z | - |
| dc.date.available | 2017-07-20 | |
| dc.date.copyright | 2015-07-20 | |
| dc.date.issued | 2015 | |
| dc.date.submitted | 2015-07-07 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/54437 | - |
| dc.description.abstract | 中文摘要
薑黃素（curcumin）為薑黃根莖中的主要色素，早期用來做食品加工的食用色素。其藥理特性也受到注目，如降血脂、抗氧化等機制也陸續有研究報導。近年研究將薑黃素歸類為組蛋白去乙醯酶（histone deacetylase）抑制劑之一，其中抗癌及毒殺細胞為主要特性。而在早些實驗室的研究中發現，將斑馬魚胚胎以薑黃素處理，發現會有尾部病變的現象。 在本研究中，我們對薑黃素引發斑馬魚肌肉病變之機制進行探討。我們發現尾部病變的比例會隨著薑黃素濃度上升，其發生速度也會因溫度提高而加快。而根據延時錄相我們發現這是一個連續性的肌肉病變現象。接著，我們利用轉基因斑馬魚（krt4:mCherry, ap:GFP）發現薑黃素會造成斑馬魚胚胎尾部的表皮和肌肉損傷，最後造成胚胎的死亡。而我們也利用質譜分析這些尾部損傷後細胞崩解釋出的蛋白。此外，在斑馬魚原位基因雜交和免疫螢光染色實驗中，我們發現經由薑黃素處理的斑馬魚胚胎尾部其層黏蛋白（laminin）的表現量下降，失去了對魚體尾部的保護，導致尾部病變的發生。我們也試著探討薑黃素分子作用機制，我們利用干擾核酸(morpholino)注射斑馬魚胚胎，降低蛋白去乙醯酶（histone deacetylase）表現，發現並沒有與薑黃素同樣的尾部潰爛現象。 最後，我們利用了許多免疫抑制劑試著抑制薑黃素引發的尾部病變現象，我們找到了海藻糖（trehalose）可以抑制尾部病變的發生，保持斑馬魚胚胎尾部完整使胚胎得以繼續存活。 | zh_TW |
| dc.description.abstract | Abstract
Curcumin (diferuloylmethane) is a pigment derived from the plant species Curcuma longa, which is frequently added in food for spice. In addition, it has been reported to be a new member of the histone deacetylase (HDAC) inhibitors and anti- cancer drug. In our previous studies, we found that curcumin functioned in inducing myopathy in zebrafish embryos during early developmental stages. In this study, our data indicated that curcumin- induced myopathy were in a dose and temperature- dependent manner. Then, a time-lapse recording was shown curcumin- induced myopathy with continuous damages. On the other hand, we used tissue specific transgenic lines to label keratinocytes and muscle cells (krt4:mCherry, ap:GFP). After the muscle damage occurred, the muscle cells and keratinocytes started to collapse and released the component proteins of the cells. We then detected these proteins in Mass analysis. The in situ hybridization and whole mount immunostaining analysis were performed to analyze this myopathy of zebrafish embryo. We found that the expression of laminin, a protein for tail bud and membrane structure, was decreased in curcumin treated embryos. To investigate possible mechanism of curcumin- induded myopathy in zebrafish embryos, we injected morpholino (MO) to knock down HDCA1 and HDAC8 expression, but it couldn’t recapitulate the phenotype of curcumin- induced myopathy. Finally, we used this zebrafish model to screen several known inhibitors, such as apoptosis inhibitors, autophagy inhibitors and immunosuppressors with the roles to inhibit the progression of curcumin-induced myopathy. Interestingly, a TOR independent autophagy inducer, inhibited curcumin- induced myopathy effectively. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T02:56:52Z (GMT). No. of bitstreams: 1 ntu-104-R02b46008-1.pdf: 1888508 bytes, checksum: e01d7133c719a941dee36f750a3472be (MD5) Previous issue date: 2015 | en |
| dc.description.tableofcontents | Content .................................................. I
中文摘要 ................................................. IV Abstract ................................................. V Introduction ............................................. 1 Curcumin ................................................. 1 Histone deacetylases (HDACs)..... ........................ 1 Myopathy...................................................3 Rapamycin and TOR .........................................4 Trehalose..................................................5 Specific aims .............................................8 Materials and methods ................................... 10 Materials ............................................... 10 Animal model............................................. 10 Culture, plasmid ........................................ 10 Drugs, enzymes, reagents ................................ 10 RNA in situ hybridization reagents .......................10 Methods ................................................. 11 Zebrafish care .......................................... 11 RNA probe preperation ................................... 12 Polymerase chain reaction (polymerase chain reaction) ... 12 DNA ligation............................................. 13 Transformation .......................................... 13 Whole-mount in situ hybridization........................ 13 Injection of morpholinos................................. 15 Whole-mount immunostaining .............................. 16 AO staining.............................................. 17 TUNEL assay.............................................. 17 Time-lapse recording and confocal images................. 18 Treatment of curcumin.................................... 18 Proteomic analysis....................................... 18 Results ................................................. 22 The phenotype and the process of curcumin induced muscle destruction or rupture in zebrafish embryos.................................................. 22 The curcumin- induced tail damage started from the tail and led to myopathy at different developmental stage.................................................... 22 The mechanism and progress in curcumin- induced myopathy zebrafish embryos. ...................................... 23 Inhibitors creening of curcumin- induced myopathy in zebrafish................................................ 25 Discussion .............................................. 26 References .............................................. 33 Figures ................................................. 40 Tables................................................... 56 | |
| dc.language.iso | en | |
| dc.title | 薑黃素引發斑馬魚肌肉病變之分子機制探討 | zh_TW |
| dc.title | Molecular mechanism of curcumin-induced myopathy in zebrafish | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 103-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 黃鵬鵬(Pung-Pung Hwang),張茂山(Mau-Sun Chang) | |
| dc.subject.keyword | 薑黃素,斑馬魚,肌肉病變,海藻糖,組蛋白去乙醯化?, | zh_TW |
| dc.subject.keyword | curcumin,zebrafish,myopathy,trehalose,histone deacetylase, | en |
| dc.relation.page | 59 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2015-07-08 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生化科學研究所 | zh_TW |
| 顯示於系所單位： | 生化科學研究所 | |
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