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DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 謝淑貞 | |
dc.contributor.author | Yu-Te Yeh | en |
dc.contributor.author | 葉昱德 | zh_TW |
dc.date.accessioned | 2021-06-17T04:30:12Z | - |
dc.date.available | 2028-08-13 | |
dc.date.copyright | 2018-08-21 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-08-13 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/70525 | - |
dc.description.abstract | 流行病學和許多研究發現長期攝取富含多酚的食物可以預防代謝異常,而中國橄欖含有大量的多酚類物質,並擁有多種生理保健功能,包含保肝、抗發炎和抗腫瘤增生等功效。本研究的第一部分,我們旨在探究中國橄欖果實甲醇萃取物-乙酸乙酯區分層(CO-EtOAc)對小鼠肝臟細胞FL83B脂質堆積的影響。結果發現CO-EtOAc可減緩油酸誘導的脂質堆積、ROS含量和MDA含量,亦由實驗結果顯示了CO-EtOAc對FL83B脂質代謝的調控機制,CO-EtOAc抑制脂肪酸轉運蛋白基因(CD36和FABP)和脂肪生成基因(SREBP-1c,FAS和ACC1),但增加了脂肪分解(HSL)和脂質氧化基因(PPARα,CPT-1和ACOX)。此外,CO-EtOAc也會增加磷酸化AMPK及ACC1,CPT-1和PPAR的蛋白質表現,但減少SREBP-1c和FAS的表達。由添加AMPK抑制劑證實其在CO-EtOAc抑制脂質堆積中扮演重要角色。因此,我們認為中國橄欖果實可以改善與脂質堆積相關的代謝異常。第二部分,我們評估CO-EtOAc對HFD和STZ誘導的糖尿病大鼠之生理調控。管餵CO-EtOAc可顯著降低體重和附睾脂肪量。此外, CO-EtOAc會增加血清HDL-C濃度、肝臟中GSH含量和抗氧化酶活性(SOD,CAT和GPx),CO-EtOAc也會改善高血糖和減少血清中TC、TG、膽酸和TNFα濃度,並減少肝臟中TC和TG的含量。我們進一步證明CO-EtOAc會抑制肝臟IRS-1磷酸化,TNF-α和IL-6表現量,但增加Akt之磷酸化。我們評估CO-EtOAc可透過調控膽固醇轉運、生物合成和降解的基因表現進而影響膽固醇代謝機制。CO-EtOAc不僅抑制SREBP-2,HMG-CoAR,SR-B1和CYP7A1的基因表現,而且還增加了膽固醇釋出轉運蛋白ABCA1和膽固醇接受器LDLR的基因表現。這些研究結果顯示,中國橄欖果實成份可以改善高脂飲食糖尿病大鼠的代謝功能失衡。第三部分,是透過 L6肌管細胞探討CO-EtOAc改善葡萄糖恆定的可能機制。CO-EtOAc會刺激葡萄糖轉運蛋白轉位至細胞膜促進葡萄糖攝取,同時增加p-GSK3α/β的蛋白質表現量進而增加肝醣儲存含量。此外,我們也發現CO-EtOAc使線粒體膜去極化、降低粒線體氧消耗並抑制粒線體NADH脫氫酶酵素活性,導致ADP / ATP比率增加,然後促使AMPK活化。最後,CO-EtOAc能夠透過活化AMPK刺激葡萄糖攝取和促進線粒體分裂,這些顯示CO-EtOAc在治療或預防HFD誘導的高血糖症中具有潛在的保健功能。總體而言,實驗結果充分顯示中國橄欖果實可做為抗肥胖和相關疾病的健康食品。 | zh_TW |
dc.description.abstract | Epidemiological numerous studies show that polyphenols contribute various beneficial effects on hepatic protection, anti-inflammation, and anti-cancer. In the first part of the study, we aimed to investigate the role of the ethyl acetate fraction of Chinese olive fruit extract (CO-EtOAc) in suppression of lipid accumulation, ROS and MDA levels in FL83B mouse hepatocytes. CO-EtOAc inhibited the mRNA levels of fatty acid transporter genes (CD36 and FABP) and lipogenesis genes (SREBP-1c, FAS, and ACC1), but upregulated genes that govern lipolysis (HSL) and lipid oxidation (PPARα, CPT-1, and ACOX). Moreover, CO-EtOAc increased the protein expression of phosphorylated AMPK, ACC1, CPT-1, and PPAR, but downregulated the expression of SREBP-1c and FAS. Adding AMPK inhibitor, attenuated the role of in CO-EtOAc-mediated amelioration of lipid accumulation. Therefore, Chinese olive fruits may have the potential to improve the metabolic abnormalities associated with hepatic lipid accumulation. The second part of the study, we evaluated the effect of CO-EtOAc on HFD and STZ-induced diabetes. CO-EtOAc could re-markedly decreased the body weight and epididymal adipose mass. In addition, CO-EtOAc increased serum HDL-C levels, hepatic GSH levels, and antioxidant enzyme activities (GSH, CAT, and GPx). Furthermore, CO-EtOAc lowered blood glucose, serum levels of TC, TG, bile acid, and TNFα; and reduced TC and TG in liver. We further demonstrated that CO-EtOAc also mildly suppressed hepatic levels of phosphorylated IRS-1, TNF-α, and IL-6, but enhanced Akt phosphorylation. The possible mechanisms of CO-EtOAc on cholesterol metabolism were assessed by determining the expression of genes involved in cholesterol transportation, biosynthesis, and degradation. It was found that CO-EtOAc not only inhibited mRNA levels of SREBP-2, HMG-CoAR, SR-B1, and CYP7A1 but also increased the expression of genes, such as ABCA1 and LDLR that governed cholesterol efflux and cholesterol uptake, respectively. Moreover, the protein expressions of ABCA1 and LDLR were also significantly increased in the liver of rats supplemented with CO-EtOAc. These findings suggest that Chinese olive fruit may improve the metabolic dysfunction in diabetic rats under HFD challenge. The third part of the study, we aimed to investigate the potential mechanisms in regulation of glucose homeostasis in L6 myotubes. CO-EtOAc effectively increased glycogen content via increased p-GSK3α/β and enhanced glucose uptake by stimulating glucose transporter 4 membraned translocation. In addition, we found that CO-EtOAc depolarized the mitochondrial membrane, decreased mitochondrial oxygen consumption and inhibited mitochondrial NADH dehydrogenase enzyme activities leading to increases in the ADP/ATP ratio, which then triggered AMPK activation. We also found CO-EtOAc is capable of stimulating glucose uptake and promoting mitochondrial fission through activation of AMPK. We thus suggest that CO-EtOAc has the therapeutic potential in the prevention of HFD-induced hyperglycemia. Overall, the experimental evidence highlight the Chinese olive fruit as a new health food in the prevention of obesity and its related metabolic disorders. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T04:30:12Z (GMT). No. of bitstreams: 1 ntu-107-D02641002-1.pdf: 4484123 bytes, checksum: a463f308eddb550aa26ab373e1ecaaed (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | 謝誌 I
摘要 II Abstract IV 目錄 VI 表目錄 IX 圖目錄 X 第一章 文獻回顧 1 第一節、肝臟中脂質代謝之調控 1 1. NAFLD的致病原因 2 2. NAFLD的致病相關機制 2 3. 肝臟脂質代謝 3 第二節、胰島素敏感性與血醣恆定之調控 7 1. 糖尿病的診斷指標與分類 8 2. 糖尿病的致病機轉 8 3. 葡萄糖攝取路徑 10 4. AMPK對糖尿病的影響 11 5. 糖尿病的動物模式 12 第三節、粒線體 (mitochondria) 13 1. 粒線體的動態平衡與其生理意義 14 2. 粒線體功能失調 (Mitochondrial dysfunction) 15 第四節、中國橄欖 16 1. 中國橄欖的成份分析 16 2. 中國橄欖的藥理學研究與相關生物活性探討 17 第二章 研究目標 20 第三章 材料與方法 21 第一節、 實驗材料 21 第二節、 實驗方法 28 1. 中國橄欖甲醇萃取-乙酸乙酯區分層(CO-EtOAc)之樣品製備 28 2. 動物實驗 28 3. 細胞培養 32 4. 細胞蛋白質萃取 33 5. 棕櫚酸和油酸之製備 36 6. 十二烷基硫酸鈉-聚丙烯醯胺凝膠電泳分析 36 7. 西方墨點法 38 8. 細胞總RNA萃取 40 9. 反轉錄 41 10. 即時定量聚合酶連鎖反應 41 11. 細胞存活率試驗 42 12. 細胞粒線體膜電位分析 42 13. 細胞粒線體壓力測試 43 14. 油紅染色 44 15. 2-NBDG 葡萄糖攝取試驗 45 16. 活性氧化物質分析 45 17. 硫代巴比妥酸值試驗 46 18. 組織學鑑定 47 19. 腺嘌呤核苷酸萃取和測定 48 20. shRNA製備與慢病毒感染 48 21. 酵素活性分析 50 22. 統計分析 51 第四章 實驗結果 52 第一節、探討中國橄欖甲醇萃取物-乙酸乙酯區分層(CO-EtOAc)對小鼠肝臟細胞FL83B脂質堆積之影響及其可能參與的脂質代謝路徑 52 1. CO-EtOAc 對小鼠肝臟細胞FL83B脂質堆積的影響 52 2. CO-EtOAc對小鼠肝臟細胞FL83B之ROS和TBARS生合成的影響53 3. CO-EtOAc減少小鼠肝臟細胞FL83B脂質堆積可能參與的途徑53 4. CO-EtOAc對小鼠肝臟細胞FL83B中AMPK和脂質代謝相關蛋白質表現量之影響 54 5. 抑制AMPK對小鼠肝臟細胞FL83B脂質堆積的影響 55 6. 活化小鼠肝臟細胞FL83B的AMPK可能路徑 55 第二節、探討中國橄欖甲醇萃取物-乙酸乙酯區分層(CO-EtOAc)對調節HFD合併STZ所誘導的高血糖與高血脂之影響 56 1. CO-EtOAc 對HFD合併STZ大鼠體重及血液生化數值的影響 56 2. CO-EtOAc 對HFD合併STZ大鼠肝臟抗氧化酵素及硫代巴比妥酸的影響 57 3. CO-EtOAc 對HFD合併STZ大鼠肝臟IRS-1和Akt的影響 58 4. CO-EtOAc 對HFD合併STZ大鼠肝臟脂質堆積的影響 58 5. CO-EtOAc對HFD合併STZ大鼠肝臟膽固醇代謝基因與蛋白質表現的影響 58 6. CO-EtOAc 對HFD合併STZ大鼠肝臟發炎反應的影響 59 第三節、探討中國橄欖甲醇萃取物-乙酸乙酯區分層(CO-EtOAc)對大鼠肌肉細胞L6葡萄糖攝取的機制 60 1. CO-EtOAc對大鼠肌肉細胞L6的葡萄糖攝取能力的影響 60 2. CO-EtOAc對大鼠肌肉細胞L6葡萄糖攝取路徑之影響 61 3. CO-EtOAc對大鼠肌肉細胞L6肝醣合成和糖解作用與之影響 61 4. CO-EtOAc對大鼠肌肉細胞L6 粒線體功能的影響 62 5. CO-EtOAc對大鼠肌肉細胞L6 粒線體分裂(fission)的影響 63 第五章 討論 65 附錄 109 參考文獻 110 | |
dc.language.iso | zh-TW | |
dc.title | 中國橄欖萃取物對脂質與醣類代謝相關機制的探討 | zh_TW |
dc.title | The study of Chinese olive (Canarium album L.) extract
on lipid and glucose metabolism | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-2 | |
dc.description.degree | 博士 | |
dc.contributor.coadvisor | 姜安娜 | |
dc.contributor.oralexamcommittee | 黃智興,郭靜娟,姜為中 | |
dc.subject.keyword | 中國橄欖,高血糖,高血脂,粒線體,葡萄醣攝取,AMPK, | zh_TW |
dc.subject.keyword | Chinese olive,hyperglycemia,hyperlipidemia,mitochondrial,glucose uptake,AMPK, | en |
dc.relation.page | 132 | |
dc.identifier.doi | 10.6342/NTU201803157 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2018-08-13 | |
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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