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DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 陳慧文(Hui-Wen Chen) | |
dc.contributor.author | Wei-Shan Chang | en |
dc.contributor.author | 張維珊 | zh_TW |
dc.date.accessioned | 2021-06-16T09:40:05Z | - |
dc.date.available | 2022-02-16 | |
dc.date.copyright | 2017-02-16 | |
dc.date.issued | 2017 | |
dc.date.submitted | 2017-02-08 | |
dc.identifier.citation | Ashfaq, U.A., Masoud, M.S., Khaliq, S., Nawaz, Z., Riazuddin, S., 2011. Inhibition of hepatitis C virus 3a genotype entry through Glanthus Nivalis Agglutinin. Virol J 8, 248.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/59828 | - |
dc.description.abstract | 中文摘要
貓傳染性腹膜炎是由貓冠狀病毒變異株所引起,於多貓的環境易發高度的盛行率以及顯著的死亡率,目前並無良好的預防及治療方式。Diphyllin為一具有阻斷宿主Vacuolar ATPase活性的化合物,已知能藉由抑制內質體酸化而干擾流感病毒與登革病毒在細胞內的感染。本研究的目的為探討diphyllin對於貓傳染性腹膜炎病毒之抗病毒效果及其作用機制,選用的病毒株為第二型的貓傳染性腹膜炎病毒NTU156。本篇論文之研究結果顯示,diphyllin能有效抑制Fcwf-4細胞的內質體酸化,並抑制病毒在細胞內的複製,具有劑量依賴效應。藥物作用時間點試驗則顯示diphyllin在病毒入侵宿主細胞之前給予之效果為最佳。另外,併用抗病毒藥物Galanthus nivalis agglutinin能夠得到更好的協同效果。而在抗體依賴增強作用的細胞感染模式之下,diphyllin仍能有效降低病毒在Fcwf-4細胞與U937細胞中的複製。與單純的藥物相比,使用奈米載體遞送之diphyllin表現出顯著較低的細胞毒性,以及較佳的病毒抑制效應。總結以上之研究結果,diphyllin對於貓傳染性腹膜炎病毒具有良好之抗病毒效果,有潛力發展為臨床使用之抗病毒藥物。 關鍵字:貓傳染性腹膜炎病毒,Diphyllin,Vacuolar ATPase 阻斷劑,抗體依賴增強作用,奈米粒子。 | zh_TW |
dc.description.abstract | Abstract
Feline infectious peritonitis (FIP), caused by a mutated feline coronavirus, is one of the most serious and fatal viral diseases in cats. So far, it remains incurable, and there is no effective vaccine commercially available. The compound diphyllin, a novel vacuolar ATPase blocker, has been shown to inhibit the intracellular endosomal acidification, thus interfering the infection of influenza virus and dengue virus. This study aims to investigate the antiviral activity of diphyllin against the type II feline infectious peritonitis virus (FIPV) strain NTU156. The results show that diphyllin dose-dependently inhibited endosomal acidification in Fcwf-4 cells. Treatment with diphyllin altered the cellular susceptibility to FIPV and inhibited the downstream virus replication. Furthermore, combinatorial treatment of the host-targeting diphyllin with pathogen-targeting Galanthus nivalis agglutinin demonstrated enhanced cell protection. In the in vitro model of antibody-dependent enhancement of FIPV infection, diphyllin also showed a significant antiviral activity against FIPV in Fcwf-4 cells and U937 cells. In addition, nanoparticulate diphyllin further demonstrated a decreased cytotoxicity and improved inhibitory effect against FIPV. These results collectively suggest that diphyllin possesses therapeutic potential for the treatment of FIP. Keywords: Feline infectious peritonitis virus, Diphyllin, Vacuolar ATPase inhibitor, Antibody-dependent enhancement, Nanoparticles | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T09:40:05Z (GMT). No. of bitstreams: 1 ntu-106-R02629004-1.pdf: 71454257 bytes, checksum: 11cb9ae25295e5fdb7f57f07889d6bb9 (MD5) Previous issue date: 2017 | en |
dc.description.tableofcontents | Contents
中文摘要 I Abstract II List of figures VI List of tables VII Chapter 1 Introduction 1 1.1. Feline coronavirus and feline infectious peritonitis virus 1 1.2. Feline infectious peritonitis: pathogenesis 3 1.3. The epidemiology of feline infectious peritonitis in Taiwan 5 1.4. Feline infectious peritonitis: diagnosis 7 1.5. Feline infectious peritonitis: treatment 8 1.6. Vacuolar ATPase (V-ATPase) and its inhibitors 9 1.7. Application of diphyllin 11 1.8. Antiviral research of Galanthus nivalis agglutinin 12 1.9. Nanoparticulate antiviral delivery system 14 1.10. Objective of this study 15 Chapter 2 Materials and methods 17 2.1. Cells, virus, and compounds 17 2.2. Measurement of in vitro cytotoxicity of diphyllin by MTT assay 18 2.3. Cell cytopathic effect (CPE) inhibition assay and determination of IC50 19 2.4. Viral plaque assay 20 2.5. Analysis of endosomal acidification with acridine orange labeling 20 2.6. Immunofluorescence staining of intracellular viral protein 21 2.7. Time-of-addition assay of diphyllin 22 2.8. Compound combinations treatment assay 22 2.9. Plasmid construction and quantitative RT-PCR (qRT-PCR) of FIPV 23 2.10. Establishment of in vitro antibody-dependent enhancement (ADE) infection of FIPV and the antiviral activity of diphyllin under the ADE infection 25 2.11. Preparation of diphyllin-loaded nanoparticles 26 2.12. Quantification of diphyllin by HPLC 26 2.13. Transmission electronic microscopy (TEM) 27 2.14. Determination of CC50 and IC50 of diphyllin-loaded nanoparticles 28 2.15. Statistical analyses 28 Chapter 3 Results 29 3.1. Cellular cytotoxicity of diphyllin 29 3.2. Dose-dependent inhibitory antiviral activity of diphyllin in Fcwf-4 cells 29 3.3. Intracellular virus inhibition activity of diphyllin by immunofluorescence 30 3.4. Diphyllin blocked endosomal acidification in Fcwf-4 cells 30 3.5. Pretreatment of diphyllin exhibits the maximal antiviral effects 31 3.6. Compound combinations of diphyllin and GNA 31 3.7. Construction of quantitative RT-PCR standard curve 32 3.8. Establishment of antibody-dependent enhancement infection models in two different cells 32 3.9. Antiviral activity of diphyllin in the context of ADE for viral infection 33 3.10. Characterization of diphyllin-loaded nanoparticles 33 3.11. Quantification of diphyllin nanoparticles by HPLC 34 3.12. Improved safety and efficacy demonstrated by nanoparticle delivery of diphyllin 35 Chapter 4 Discussion 36 Chapter 5 Figures and tables…………………………………………………………...41 Chapter 6 References………………………………………………………….……….60 | |
dc.language.iso | en | |
dc.title | Vacuolar ATPase阻斷劑Diphyllin抑制貓傳染性腹膜炎病毒之效果 | zh_TW |
dc.title | Inhibitory effect of diphyllin, a vacuolar ATPase blocker, on feline infectious peritonitis virus | en |
dc.type | Thesis | |
dc.date.schoolyear | 105-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 王金和(Ching-Ho Wang),張惠雯(Hui-Wen Chang),胡哲銘(Che-Ming Jack Hu),張麗冠(Li-Kwan Chang) | |
dc.subject.keyword | 貓傳染性腹膜炎病毒,diphyllin,vacuolar ATPase 阻斷劑,抗體依賴增強作用,奈米粒子, | zh_TW |
dc.subject.keyword | Feline infectious peritonitis virus,Diphyllin,Vacuolar ATPase inhibitor,Antibody-dependent enhancement,Nanoparticles, | en |
dc.relation.page | 65 | |
dc.identifier.doi | 10.6342/NTU201600143 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2017-02-08 | |
dc.contributor.author-college | 獸醫專業學院 | zh_TW |
dc.contributor.author-dept | 獸醫學研究所 | zh_TW |
顯示於系所單位: | 獸醫學系 |
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