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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 吳信志(Shinn-Chih Wu) | |
| dc.contributor.author | Tzu-Jou Chen | en |
| dc.contributor.author | 陳子柔 | zh_TW |
| dc.date.accessioned | 2022-11-24T03:02:52Z | - |
| dc.date.available | 2021-07-23 | |
| dc.date.available | 2022-11-24T03:02:52Z | - |
| dc.date.copyright | 2021-07-23 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-07-11 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/80227 | - |
| dc.description.abstract | "心血管疾病是造成全球人口死亡之主要疾病之一，其中又以心肌梗塞占多數，在冠狀動脈阻塞後，心臟組織缺氧導致心肌細胞及心臟成纖維細胞受損，進而引發炎症及心臟重塑，將使心臟增加10%至35%之纖維組織，可能導致心臟功能受損、心律不整及收縮障礙，甚至心臟衰竭，因此探討心肌梗塞（myocardial infarction, MI）後如何維持心臟功能是刻不容緩的重要議題。 近年來許多研究開始關注細胞受損後所釋放之危險相關分子模型（danger associated molecular patterns, DAMPs）之訊息傳遞與調控，DAMPs包含HMGB1、S100A1/A8/A9及IL-1α等，具有活化常駐型巨噬細胞及增加內皮細胞通透性功能，其可招募嗜中性球及單核球使發炎反應加劇，活化成纖維細胞使其增生並累積細胞外基質。前人研究發現以S100A8/A9蛋白質預處理之人類間葉幹細胞（human mesenchymal stem cell, hMSC）可加速傷口癒合、降低疤痕組織，並對心臟具有保護作用。因此，本研究旨在探討經S100A8/A9 預處理的 hAMSCs條件培養液是否於 MI 後發揮心臟保護作用並維持心臟功能。 本研究分為體內試驗及體外試驗兩部分，第一階段，體外試驗將分別分離胎鼠心肌細胞（neonatal cardiomyocytes, nCMs）及人類羊膜間葉幹細胞（human amniotic mesenchymal stem cells, hAMSCs）進行初代培養與細胞特性分析作為本試驗細胞來源，再利用nCMs特異性磁株抗體純化後誘導其缺氧/復氧損傷（hypoxia/ reoxygenation, H/R），並以RT-PCR、qPCR進行檢測，與對照組相比，誘導H/R損傷之nCM處理組之S100蛋白、IL-1、IL-6、TNF-α和IFNγ等細胞受損、促纖維化增生及促炎症相關基因表達具有顯著差異。而人類羊膜間葉幹細胞則以含有額外添加S100A8/A9之培養液與hAMSCs培養，後以RT-PCR、qPCR確認細胞是否產生相應之基因表現，結果顯示以S100A8/A9預處理之hAMSC則於免疫調節和組織修復相關基因，包含TLR、CCR7、IL-10、GDF9、SPARCKL、MMP、TIMP等與對照組相比亦有顯著變化。此外，此研究通過在缺氧條件下更換受損 nCM 之培養液檢測與控制組相比經預處理之hAMSC條件培養液是否具有保護作用，結果顯示hAMSC來源之條件培養液組之細胞存活率顯著高於基礎培養基組，然而，未預處理之hAMSC組與S100A8/A9預處理之hAMSC組之間沒有顯著差異，除此之外，各組在某些基因表達水平之上調和下調結果則顯示hAMSC於預處理後會產生適應性反應，並調控相關之分子機制。 第二階段體內試驗，使用8週齡C57BL/6小鼠心肌梗塞/再灌流模型，以靜脈注射經S100A8/A9預處理後之hAMSC條件培養液進行治療。試驗將分為五組，分別為控制組、假手術組、伪治療組（基礎培養液）、一般治療組（hMSC條件培養液）及預處理治療組（經S100A8/A9預處理後之hMSC條件培養液），於治療後3天、1週、2週及4週以血清學檢查、心臟超音波、組織切片染色檢測心臟結構、功能與血管密度之變化，於心臟超音波結果顯示，預處理之hAMSC組之左心室射血分數和收縮分率顯著高於未預處理hAMSCs組和I/R組（P<0.05），纖維化區域之組織切片染色定量分析，亦顯示各組間纖維化具有顯著差異，並且預處理之hAMSC組雖與一般治療組並無統計上之顯著差異，然其纖維組織之定量結果仍為各處理中最少之組別。 總結，我們證實 S100A8/A9 預處理的 hAMSCs 條件培養基在體外研究和小鼠心肌 I/R 模型的治療中具有增強之修復潛力，期望此研究結果有益於受損後心臟功能之維持，為未來的心肌梗塞相關研究提供良好的治療方向與展望並貢獻於臨床治療。 " | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T03:02:52Z (GMT). No. of bitstreams: 1 U0001-0907202117240900.pdf: 3343940 bytes, checksum: 4f8537031914afe75a840b33e8a4f870 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | 口試委員會審定書 謝誌…………………………Ⅰ 摘要…………………………Ⅱ ABSTRACT…………………………Ⅳ 目錄…………………………Ⅶ 圖次…………………………Ⅹ 表次…………………………ⅩⅠ ABBRREVIATIONS…………………………ⅩⅡ 第一章 緒論…………………………1 第二章 文獻探討…………………………3 2.1 心臟基本構造…………………………3 2.1.1 心臟解剖構造…………………………3 2.1.2 心臟組成及細胞類型…………………………5 2.1.3 心臟生理功能…………………………5 2.2 心臟疾病…………………………7 2.2.1 心肌梗塞…………………………8 2.2.2 心臟疾病體外細胞模型…………………………9 2.2.3 心臟疾病體內動物模型…………………………11 2.3 心臟治療…………………………13 2.4 幹細胞…………………………16 2.4.1 幹細胞定義與分類…………………………16 2.4.2 幹細胞治療…………………………18 2.4.3 羊膜間葉幹細胞…………………………19 2.5 預處理間葉幹細胞之治療策略…………………………21 2.5.1 預處理間葉幹細胞之治療策略綜述…………………………21 2.5.2 S100鈣離子結合蛋白家族之功能…………………………22 2.5.3 S100A8與間葉幹細胞之相互作用…………………………24 第三章 試驗研究…………………………26 3.1 人類羊膜間葉幹細胞之體外分離培養與S100A8/A9預處理系統之建立…………………………26 3.1.1 前言…………………………26 3.1.2 試驗設計與材料方法…………………………27 3.1.3 試驗結果與討論…………………………35 3.2 小鼠心肌細胞體外培養及誘導低氧/復氧損傷模型與人類羊膜間葉幹細胞條件培養液之體外治療潛能…………………………43 3.2.1 前言…………………………43 3.2.2 試驗設計與材料方法…………………………44 3.2.3 試驗結果與討論…………………………50 3.3 小鼠心肌梗塞結紮與再灌流疾病動物模型之建立與人類羊膜間葉幹細胞條件培養液之治療潛能…………………………57 3.1.1 前言…………………………57 3.1.2 試驗設計與材料方法…………………………58 3.1.3 試驗結果與討論…………………………62 第四章 綜合討論…………………………70 第五章 結論…………………………73 第六章 未來展望…………………………75 第七章 參考文獻…………………………76 | |
| dc.language.iso | zh-TW | |
| dc.subject | S100A8/A9 | zh_TW |
| dc.subject | 人類羊膜間葉幹細胞條件培養液 | zh_TW |
| dc.subject | 小鼠心肌梗塞/再灌流模型 | zh_TW |
| dc.subject | S100A8/A9 | en |
| dc.subject | murine myocardial I/R model | en |
| dc.subject | human amniotic mesenchymal stem cell derived condition medium | en |
| dc.title | S100A8/A9預處理於人類羊膜間葉幹細胞條件培養液對心肌梗塞修復潛能之探討 | zh_TW |
| dc.title | Repairing myocardial infarction with S100A8/A9 pretreatment in conditioned medium from human amniotic membrane mesenchymal stem cells | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 宋麗英(Hsin-Tsai Liu),陳全木(Chih-Yang Tseng),李愛先,彭劭于 | |
| dc.subject.keyword | 人類羊膜間葉幹細胞條件培養液,小鼠心肌梗塞/再灌流模型,S100A8/A9, | zh_TW |
| dc.subject.keyword | human amniotic mesenchymal stem cell derived condition medium,murine myocardial I/R model,S100A8/A9, | en |
| dc.relation.page | 92 | |
| dc.identifier.doi | 10.6342/NTU202101369 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-07-12 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 動物科學技術學研究所 | zh_TW |
| 顯示於系所單位： | 動物科學技術學系 | |
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