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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 蔡沛學(Pei-Shiue Tsai) | |
dc.contributor.author | Hung-Ting Liu | en |
dc.contributor.author | 劉洪廷 | zh_TW |
dc.date.accessioned | 2021-06-17T01:58:06Z | - |
dc.date.available | 2020-08-29 | |
dc.date.copyright | 2017-08-29 | |
dc.date.issued | 2017 | |
dc.date.submitted | 2017-07-20 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/67919 | - |
dc.description.abstract | Cisplatin 或 cis-diamminedichloroplatinum (II)是一種常用的抗癌藥物,從 1970 年起被使用於治療許多癌症,它能有效的治療頭頸癌、肺癌、膀胱癌、生殖細胞 癌等,然而,此藥物所導致的腎毒性副作用卻時常限制了它於臨床上的使用。單 次注射 Cisplatin 造成的腎毒性或急性腎衰竭之發生率大約為 20%-30%,而且,重 複地施打 Cisplatin 也會加重其造成的腎臟損傷,因此,在癌症治療的過程中,時 常不得不停止使用 Cisplatin。雖然 Cisplatin 造成之腎毒性的詳細機制尚不清楚, 但目前證據指出主要可能與 Cisplatin 進入細胞後產生的氧化性傷害以及後續引起 的發炎反應有關。 異厚圤酚(Honokiol)是一種由 Magnolia obovate 或 Magnolia officinalis 的樹 幹所萃取出來的多酚類化合物,它具有多種生物活性,包括抗氧化、抗過敏、抗 焦慮、抗癌、抗憂鬱以及神經保護作用,近年來的許多離體 (in vitro) 及活體 (in vivo) 實驗也證實異厚圤酚具有抗發炎以及抗氧化的作用。 本研究藉由 Cisplatin 誘導的腎衰老鼠與 Madin-Darby 犬腎臟上皮細胞來探討 異厚圤酚是否具有保護腎臟的效用,我們使用多種不同的實驗設計來探討當腎臟 細胞暴露在 Cisplatin 或異厚圤酚中時,細胞骨架(微絲及微管)以及兩種維持細 胞極性的膜蛋白(E-cadherin 以及 Occludin)之間的變化與關係。首先,我們發 現 Cisplatin 會造成 E-cadherin 及 Occludin 的位置改變,再者,我們首次發現異厚 圤酚可以穩定 E-cadherin 及 Occludin 在細胞中的位置,防止它們受到 Cisplatin 的 影響,另外,異厚圤酚會增加聚合微絲以及微管在腎臟上皮細胞中的數量,這可 能與它能夠穩定 E-cadherin 以及 Occludin 在細胞位置的機制有關,在動物實驗 中,異厚圤酚則能減緩 Cisplatin 造成的腎毒性,它不僅維持了腎臟的基本機能以及構造,同時也減少間質性的纖維化與炎症反應,雖然我們期望和厚圤酚能在細 胞實驗產生抗氧化以及抗細胞凋亡的作用,但是在使用免疫染色法觀察氧化緊迫 指標蛋白 8-OHdG 以及細胞凋亡指標蛋白 Caspase-3 的變化時卻未能查覺此現 象。綜合本研究的結果,異厚圤酚可能具有減少 Cisplatin 造成的腎臟細胞毒性以 及器官毒性之療效,因此異厚圤酚非常有機會作為與 Cisplatin 共同給予癌症病患的藥物。 | zh_TW |
dc.description.abstract | Cisplatin or cis-diamminedichloroplatinum (II) has been used as an anti-cancer drug since 1970s. It is proven to be an effective therapeutic agent in treating head, neck, lung, bladder, and germ cell cancers. Unfortunately, the accompanied nephrotoxicity compromises its wide applications at clinics. The incidence of Cisplatin-induced nephrotoxicity or acute kidney injury ranges from 20%-30% following a single administration. Furthermore, the severity of Cisplatin- induced nephrotoxicity increases with repeated treatments/doses, thus often impede the cancer treatment. Although the exact mechanism that is responsible for Cisplatin-induced nephrotoxicity is not fully understood; however, the induction of oxidative stress and the activation of inflammatory responses might be involved. Honokiol, a polyphenol constituent extracted from the bark of Magnolia obovate or Magnolia officinalis, exhibits several important bioactivities, including anti-oxidation, anti-allergy, anti-anxiety, anti-cancer, anti-depression, and neuroprotection. Moreover, recent studies showed that Honokiol exhibits anti-inflammation and antioxidant effects in numbers of in vitro and in vivo models. In this study, we applied in vivo Cisplatin-induced kidney injury mouse model and in vitro Madin-Darby Canine Kidney Epithelial Cells (MDCK) cell culture-based analyses to investigate whether Honokiol exhibits the protective effects on kidney function. By applying various independent techniques, we investigated the relationship and changes between the cytoskeletons (actin and tubulin), and two important cell polarity maintaining proteins, E-cadherin and Occludin, upon Cisplatin and/or Honokiol challenges. First we demonstrated that Cisplatin can disrupt the localization of E-cadherin and Occludin in MDCK cells. Second, we disclosed a new property of Honokiol on its ability to stabilize cellular junction proteins against Cisplatin-induced alteration. Furthermore, Honokiol slightly increased the amount of F-actin and expended the structure of microtubule, which likely contributed to the maintenance of cellular localization of E-cadherin and Occludin at the cellular junctions. More importantly, Honokiol attenuated Cisplatin-induced nephrotoxicity in our animal model. Honokiol treatment not only preserved the basic kidney functions and structures, but also reduced the interstitial fibrosis and inflammation in general. Although we expected that Honokiol achieved its protective effect by its anti-oxidation and anti-apoptosis abilities, but immunolabeling of oxidative stress marker protein 8-hydroxy-deoxy-guanidine (8-OHdG) and apoptosis marker Caspase-3 didn’t fully support our speculations. Nevertheless, based on our results, Honokiol seems to be a promising candidate to be given along with Cisplatin to reduce the cyto-/ organ toxicity of Cisplatin and to improve the clinical safety of Cisplatin usage in cancer patients. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T01:58:06Z (GMT). No. of bitstreams: 1 ntu-106-R04629005-1.pdf: 31465127 bytes, checksum: 5e3a0512eecc1183b2ab1e18149ab30a (MD5) Previous issue date: 2017 | en |
dc.description.tableofcontents | 口試委員會審定書................................................................................................i 誌謝.................................................................................................ii 中文摘要..........................................................................................iii Abstract...............................................................................................v Contents..............................................................................................vii List of Figures..................................................................................x List of Tables...................................................................................xi Chapter 1 Introduction...........................................................................1 1.1 General kidney structure and its physiological functions......................1 1.2 The importance of epithelial cell polarity for kidney functions...........................2 1.3 Kidney injury................................................................................4 1.3.1 Clinical signs..........................................................................4 1.3.2 Pathological findings.....................................................................4 1.3.3 Disruptions of critical components/proteins at the cellular level.....................5 1.4 Introduction of Cisplatin............................................................6 1.4.1 Cellular actions of Cisplatin..............................................................6 1.4.2 Side effects and alternative platinum compounds...........................................7 1.4.3 Know causes of Cisplatin-induced nephrotoxicity..........................8 1.4.4 Pharmacokinetics of Cisplatin..................................................10 1.5 Anti-oxidant........................................................................10 1.5.1 Source of oxidative stress........................................................10 1.5.2 Anti-oxidants........................................................................11 1.6 Honokiol...................................................................................12 1.6.1 Known functions.....................................................................12 1.6.1.1 Anti-inflammation...............................................................13 1.6.1.2 Anti-oxidative stress.............................................................13 1.6.1.3 Anti-cancer............................................................................13 1.6.2 Other natural derivatives with similar anti-inflammation or anti-oxidation effects...................................................................................14 1.6.2.1 Curcumin......................................................................14 1.6.2.2 Ginger extract...............................................................15 1.7 The scope of this thesis............................................................15 Chapter 2 Materials and Methods.......................................................17 2.1 Chemicals, reagents, antibodies....................................................17 2.2 Cell culture..............................................................................17 2.3 2D polarized Transwell® culture.......................................................18 2.4 Cell viability assay (MTT assay)...........................................................19 2.5 Cell spreading assay......................................................................19 2.6 Cytoskeleton (F-actin and tubulin) quantification assay...................................20 2.7 Immuno-blotting.............................................................................21 2.8 Indirect immunofluorescence staining and image acquisition..........................22 2.9 Correlation analysis of cell junction proteins Occludin and E-cadherin in MDCK cells..............................................................................................23 2.10 Oxidative stress and apoptosis evaluation............................................24 2.11 Animal, metabolic cage experiments and serum, urine analysis......................25 2.12 Morphologic and pathological evaluation of kidney tissue.............................28 Chapter 3 Results.........................................................................30 3.1 Concentration determination of Cisplatin and Honokiol...................................30 3.2 Cisplatin inhibited cell spreading....................................................31 3.3 Honokiol attenuated Ciplatin-induced disorganization of Occludin and E-cadherin........................................................................................33 3.4 Cisplatin caused vertical movement of Occludin and lateral movement of E-cadherin, and Honokiol partially maintained the localization of Occludin but not E-cadherin........................................................................................35 3.5 Honokiol increased the amount of F-actin and tubulin bundle in MDCK cells..39 3.6 Honokiol did not reduce the formulation of 8-OHdG caused by Cisplatin........39 3.7 Honokiol had no effect on the increased Caspase-3 expression induced by Cisplatin..............................................................................39 3.8 Honokiol attenuated Cisplatin induced nephrotoxicity in ICR mice..................42 3.8.1 Renal function.........................................................42 3.8.2 Kidney morphology evaluation........................................47 Chapter 4 Discussions............................................................51 Chapter 5 Conclusions and future work..........................................60 References..........................................................................61 | |
dc.language.iso | en | |
dc.title | 以 Cisplatin 誘導腎毒性之小鼠為動物模式探討異厚圤酚對細胞骨架及細胞極性之影響 | zh_TW |
dc.title | Investigate the Effects of Honokiol on Cisplatin-Induced Nephrotoxicity Mouse Model: From Cytoskeleton Alterations to Cell Polarity Maintenance | en |
dc.type | Thesis | |
dc.date.schoolyear | 105-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 詹東榮,李雅珍,周濟眾,陳明汝 | |
dc.subject.keyword | Cisplatin,異厚?酚,細胞極性,腎毒性,細胞骨架, | zh_TW |
dc.subject.keyword | Cisplatin,Honokiol,Cell polarity,Nephrotoxicity,Cytoskeleton, | en |
dc.relation.page | 68 | |
dc.identifier.doi | 10.6342/NTU201701728 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2017-07-20 | |
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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