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研究生:吳曼如
研究生(外文):Wu Man Ru
論文名稱:樟芝子實體主要揮發性成分2,4,5-trimethoxybenzaldehyde對於3T3-L1細胞之影響
論文名稱(外文):Effect of 2,4,5-trimethoxybenzaldehyde,the Major Volatile Component of Antrodia camphorate Fruiting Body, on 3T3-L1 Adipocytes
指導教授:郭家芬郭家芬引用關係
指導教授(外文):Kuo, Chia-Feng
口試委員:潘敏雄陳勁初
口試委員(外文):Pan, Min-HsiungChen, Chin-Chu
口試日期:2012-01-30
學位類別:碩士
校院名稱:實踐大學
系所名稱:食品營養與保健生技學系碩士班
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:114
中文關鍵詞:245-TMBA脂質合成3T3-L1 細胞
外文關鍵詞:2,4,5-TMBAadipogenesis3T3-L1 cells
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肥胖已被認為是許多慢性疾病的危險因子,如高血壓、糖尿病及動脈粥狀硬化等。樟芝(Antrodia camphorata)之子實體已被證實具抗氧化、降血壓、抗癌、降血脂及保肝等功效,但樟芝對於體脂堆積的影響尚未被探討。因樟芝子實體中主要揮發性成分2,4,5-TMBA具有抑制COX-2活性,而COX-2在脂肪細胞分化中扮演了重要的角色,本實驗探討2,4,5-TMBA對於3T3-L1脂肪前驅細胞的分化及成熟脂肪細胞脂解能力之影響。以紅油染色方法顯示在分化階段,2,4,5-TMBA會隨著劑量的提升而抑制脂肪的形成。在分子層次上,2,4,5-TMBA會抑制參與細胞訊息傳遞路徑的MEK、ERK、JNK 及參與脂肪細胞中主要的轉錄因子 C/EBPs、PPAR γ、ADD1/SREBP1c 並抑制參與脂肪酸合成的acetyl-CoA carboxylase及脂肪細胞表面蛋白perilipin A的蛋白質表現量。當2,4,5-TMBA處理成熟脂肪細胞時,會提升荷爾蒙敏感性脂解酶HSL 及降低 perilipin A的蛋白質表現,而提升三酸甘油酯被水解的能力。綜合以上的結果顯示,2,4,5-TMBA會抑制3T3-L1脂肪前驅細胞的分化並促進成熟脂肪細胞內的脂質分解。

關鍵字: 2,4,5-TMBA、脂質合成、脂解能力、3T3-L1脂肪細胞

Obesity is considered a major risk factor for many chronic disease, including hypertension, type 2 diabetes, and atherosclerosis.The fruiting bodies of Antrodia camphorate possess various biological activites, including anti-oxidation, anti-hypertension, anti-carcinogenesis, anti-hyperlipidemia and liver-protection.
However, there has not been any study related to anti-obesity. The major volatile compound of Antrodia camphorate fruiting body, 2,4,5-trimethoxybenzaldehyde (2,4,5-TMBA), was reported to be a significantly inhibitor of COX-2 activity. Because COX-2 is essential for adipogenesis, we investigated the effect of 2,4,5-TMBA on 3T3-L1 preadipocytes differentiation and adipocytes lipolysis. During differentiation, 2,4,5-TMBA dose-dependently attenuated lipid accumulation in differentiated adipocyte as evidenced by Oil Red O staining. At the molecular level, 2,4,5-TMBA decreased the protein expression of MAP kinase kinsae (MEK), extracellular signal-regulated kinase (ERK), c-Jun amino-terminal kinase (JNK), CCAAT/enhancer binding protein (C/EBP) α,β,γ, peroxisome proliferator-activated receptor (PPAR) γ, adipocyte determination and differentiation-dependent factor1/sterol regulation element binding protein1c (ADD1/SREBP1c), acetyl-CoA carboxylase (ACC), and perilipin A. In mature adipocytes, 2,4,5-TMBA singnificantly decreased intracellular lipid accumulation by increasing the lipolysis of triglyceride though suppression of perilipin A and up-regulation of hormone-sensitive lipase (HSL). These results demonstrate that 2,4,5-TMBA efficiently inhibits adipogenesis and increase lipolysis in 3T3-L1 cells.

Keywords:2,4,5-TMBA, adipogenesis, lipolysis, 3T3-L1 cells

內容 頁次
審查通過證明書 I
博碩士論文電子檔案上傳授權書 II
致謝 III
目錄 IV
圖目錄 VI
表目錄 VII
摘要 VIII
Abstract XI
前言 1
壹、脂肪組織 2
貳、脂肪細胞之研究 5
一、脂肪前驅細胞之細胞株 5
二、脂肪前驅細胞分化過程 6
三、脂肪細胞脂質代謝 14
参、參與脂肪細胞分化及代謝相關基因之介紹 18
一、PPAR家族 18
二、CCAAT-增強子結合蛋白(C/EBPs) 20
三、環氧化酶(COX) 22
肆、樟芝 23
伍、實驗假說 28
材料與方法 29
一、實驗設計 29
二、樣品來源及配置 31
三、細胞之培養 31
四、誘導3T3-L1脂肪細胞分化 33
五、細胞毒性試驗(MTT assay) 36
六、測量脂質堆積情形 37
(一)油紅染色測定 37
(二)三酸甘油酯含量 40
七、脂肪細胞激素 41
(一)脂聯素 41
(二)瘦體素 42
八、成熟脂肪細胞lipolysis之測定(甘油釋放濃度) 43
九、原漿微粒蛋白質之分離 44
十、蛋白質定量 45
十一、電泳分析 45
十二、西方點墨法 47
十三、統計與分析 47
結果 53
一、2,4,5-TMBA對於脂肪前驅細胞分化之影響 53
(一)細胞毒殺性 53
(二)油紅染色測定 55
(三)三酸甘油酯含量測定 55
(四)MAPK蛋白質之表現 59
(五)脂肪細胞轉錄因子蛋白質之表現 62
(六)脂質合成相關蛋白質之表現 66
二、2,4,5-TMBA對於成熟脂肪細胞之影響 69
(一)油紅染色測定 69
(二)三酸甘油酯含量測定 69
(三)成熟脂肪細胞釋放甘油之含量 75
(四)成熟脂肪細胞分泌脂肪激素 75
(五)脂肪分解相關蛋白質之表現 79
(六)脂肪激素蛋白質之表現 80
討論 82
結論 94
文獻回顧 96




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