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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 謝淑貞 | |
| dc.contributor.author | Hsin-Ya Lin | en |
| dc.contributor.author | 林欣雅 | zh_TW |
| dc.date.accessioned | 2021-06-14T17:10:56Z | - |
| dc.date.available | 2016-09-20 | |
| dc.date.copyright | 2011-09-20 | |
| dc.date.issued | 2011 | |
| dc.date.submitted | 2011-08-22 | |
| dc.identifier.citation | 行政院衛生署。2010。中華民國98年國人死因統計結果分析。
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/40997 | - |
| dc.description.abstract | 代謝症候群(metabolic syndrome)盛行於已開發國家中，為數種代謝異常症狀的綜合臨床徵兆。根據前人研究，龍眼花水萃物於高果糖餵食誘導模式中可減緩大鼠之代謝症候群症狀，然而，其分子機制路徑尚未得知。因此，本實驗室取基礎飲食組(A)、高果糖飲食組(B)、龍眼花水萃物灌食組(C)之大鼠肝臟進行蛋白質體學方法分析，利用差異性螢光標記二維電泳膠體(2-dimensional differential gel electrophoresis, 2D-DIGE)及電噴霧串聯質譜法（electrospray ionization-tandem mass spectrometry, ESI-MS-MS）鑑定出在三種實驗條件下表現量具有差異的蛋白質，透過文獻探討與實驗確認，試圖建立各蛋白質間所依循的分子路徑，提出龍眼花水萃物對於代謝症候群改善作用之假說。本實驗室在分析蛋白質體學資料後，發現數種具有結合細胞內鈣離子能力且受內質網壓力所調控之蛋白質，分別為78 kDa glucose-regulated protein (GRP78)、94 kDa glucose-regulated protein (GRP94)、calreticulin (CRT)、 protein disulfide-isomerase A3 (PDIA3)，其中GRP78蛋白被眾多文獻用於作為偵測內質網壓力之指標；數據已透過即時定量聚合酶連鎖反應作進一步確認，因此我們推斷龍眼花水萃物可能透過抑制內質網壓力來減緩代謝症候群。以HepG2細胞模式進行驗證，結果顯示龍眼花水萃物可抑制tunicamycin所誘發的內質網壓力，使上述四種蛋白質表現量降低，證明龍眼花水萃物具有抑制內質網壓力之效果，而可佐證龍眼花水萃物改善代謝症候群之分子機制可能部分來自抑制內質網壓力。 | zh_TW |
| dc.description.abstract | Metabolic syndrome (MetS) is prevalent in the developed countries, consisting of several metabolic abnormal symptoms. According to the previous study, the water extract of Longan (Dimocarpus longan Lour.) flower (LFWE) could attenuate the symptoms of MetS in fructose-fed rat model. Nevertheless, the molecular mechanism remains unknown. In this study, we employed proteomic analysis by 2-dimensional differential gel electrophoresis (2D-DIGE) and electrospray ionization-tandem mass spectrometry (ESI-MS-MS) to investigate the possible molecular pathway. Through the differential protein patterns displayed in the livers among control, high fructose, and high fructose plus LFWE treated rats, the mechanism underlying the preventive function of LFWE and the pathogenic signaling pathways MetS might be explored. Through proteomic results, we have identified the differential expression of several endoplasmic reticulum (ER) stress related proteins, including 78 kDa glucose-regulated protein (GRP78), 94 kDa glucose-regulated protein (GRP94), calreticulin (CRT), protein disulfide-isomerase A3 (PDIA3). The real time PCR results further confirmed the involvement of the above proteins after LFWE treatment. We thus hypothesize that endoplasmic reticulum(ER) stress may be the target for LFWE mediated prevention of MetS. Utilizing HepG2 cell as cell model, we observed that LFWE inhibited ER stress and the mRNA expression of those ER stress marker proteins identified from our proteomics results. The result indicates LFWE might attenuate MetS at least partially through inhibiting ER stress. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-14T17:10:56Z (GMT). No. of bitstreams: 1 ntu-100-R98641036-1.pdf: 24670292 bytes, checksum: eaf904219a824c8d85fdd650096ff53c (MD5) Previous issue date: 2011 | en |
| dc.description.tableofcontents | 摘要 ii
Abstract iii 目錄 viii 壹、前言 1 貳、文獻回顧 3 一、代謝症候群(metabolic syndrome)於臺灣盛行之現況 3 二、代謝症候群(metabolic syndrome)之成因 5 （一） 研究歷史 5 （二） 症狀定義 5 （三） 病理成因 5 三、高果糖飲食誘導代謝症候群(metabolic syndrome)之動物模式 13 （一） 果糖之消化吸收作用 13 （二） 果糖於肝臟之代謝路徑 13 （三） 攝取過量果糖導致體內代謝異常之效應 15 四、龍眼花水萃物(LFWE)之先期研究成果 20 （一） 龍眼花 20 （二） 龍眼花萃取物功效成分研究 20 五、蛋白質體學分析技術之應用 21 （一） 蛋白質體 21 （二） 蛋白質體學研究方法 22 參、研究目的與實驗架構 25 一、研究目的 25 二、實驗架構 25 肆、材料與方法 27 一、實驗材料來源 27 （一） 大鼠肝臟 27 （二） HepG2細胞 27 （三） 試劑與藥品 28 二、實驗方法 29 （一） 蛋白質體學分析技術 29 （二） 即時定量聚合酶連鎖反應(real-time PCR) 32 （三） 細胞試驗 37 伍、結果 39 一、蛋白質體學差異性螢光標記二維膠體電泳(2D-DIGE) 39 （一） 基礎飲食組(A)與高果糖飲食組 (B)比較結果(spot 1~115) 41 （二） 高果糖飲食組(B)與龍眼花水萃物灌食組(C)比較結果(spot 116~199) 47 二、電噴霧串聯質譜法(ESI-MS-MS)鑑定結果 53 （一） 於高果糖飲食組(B)表現量上升之蛋白質（相較於基礎飲食組(A)） 53 （二） 於高果糖飲食組(B)表現量下降之蛋白質（相較於基礎飲食組(A)） 58 （三） 於龍眼花水萃物灌食組(C)表現量上升之蛋白質（相較於高果糖飲食組(B)） 62 （四） 於龍眼花水萃物灌食組(C)表現量下降之蛋白質（相較於高果糖飲食組(B)） 66 三、資料探勘──建立蛋白質連結(correlation mapping) 71 （一） 龍眼花水萃物(LFWE)於醣類代謝之影響 71 （二） 龍眼花水萃物(LFWE)於脂肪代謝、合成之影響 75 （三） 龍眼花水萃物(LFWE)於胰島素訊息傳導、分泌之可能影響 78 （四） 龍眼花水萃物(LFWE)對於蛋白質代謝之影響──氨甲醯磷酸合成酶(CPS1) 83 （五） 龍眼花水萃物(LFWE)對於氧化還原恆定之影響──過氧化物酶-1 (PRDX1)、蛋白質雙硫鍵異構酶(PDI) 84 （六） 龍眼花水萃物(LFWE)對於鈣離子結合及蛋白質摺疊作用之影響──GRP78、GRP94、CRT、PDIA3、RGN 86 四、參與未摺疊蛋白質反應(UPR)相關蛋白(GRP78、GRP94、CRT、PDIA3)之real-time PCR結果 93 （一） 大鼠肝臟組織中GRP78、GRP94、CRT、PDIA3 mRNA表現量比較結果 93 （二） 誘發HepG2細胞產生內質網壓力(ER stress)模式下龍眼花水萃物(LFWE)對於GRP78、GRP94、CRT、PDIA3 mRNA表現量之影響 98 陸、討論 111 一、大鼠肝臟組織real-time PCR結果 111 二、龍眼花水萃物(LFWE)之抗氧化能力可能透過調控內質網壓力(ER stress)展現 113 三、Sirtuins可能是龍眼花水萃物(LFWE)影響相關蛋白其表現量之上游因子 114 （一） Sirt1透過SREBP調控脂肪合成酶(FAS) 114 （二） Sirt1可能透過forkhead box O1 (FoxO1)調控GRP78表現，並與內質網壓力(ER stress)存在關聯性 115 （三） Sirt1可能透過調控β-catenin而影響RGN表現 116 （四） Sirt4與Sirt5各調控穀胺酸脫氫酶(GDH1)及氨甲醯磷酸合成酶(CPS1) 117 四、胰島素作用於中樞神經系統與肝臟之連結 119 柒、結論 121 捌、參考文獻 123 玖、附錄 135 | |
| dc.language.iso | zh-TW | |
| dc.subject | 未摺疊蛋白質反應 | zh_TW |
| dc.subject | 代謝症候群 | zh_TW |
| dc.subject | 高果糖飲食 | zh_TW |
| dc.subject | 龍眼花水萃物 | zh_TW |
| dc.subject | 蛋白質體學 | zh_TW |
| dc.subject | 內質網壓力 | zh_TW |
| dc.subject | high-fructose diet | en |
| dc.subject | unfolded protein response | en |
| dc.subject | ER stress | en |
| dc.subject | 2D-DIGE | en |
| dc.subject | metabolic syndrome | en |
| dc.subject | longan flower water extract | en |
| dc.title | 於高果糖餵食誘導模式下以蛋白質體學方法探討龍眼花水萃物改善大鼠代謝症候群之機制 | zh_TW |
| dc.title | The Proteomic Based Mechanism Study Governing the Longan (Dimocarpus longans Lour.)Flower Water Extract-mediated Prevention of Metabolic Syndrome in A High Fructose-fed Rat | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 99-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 廖辰中,吳亮宜,羅翊禎,高承福 | |
| dc.subject.keyword | 代謝症候群,高果糖飲食,龍眼花水萃物,蛋白質體學,內質網壓力,未摺疊蛋白質反應, | zh_TW |
| dc.subject.keyword | metabolic syndrome,high-fructose diet,longan flower water extract,2D-DIGE,ER stress,unfolded protein response, | en |
| dc.relation.page | 162 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2011-08-22 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 食品科技研究所 | zh_TW |
| 顯示於系所單位： | 食品科技研究所 | |
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