四角菱角殼萃取物對脂肪細胞脂質生合成和肌管細胞粒線體功能之影響及其分子機轉研究

Effects of Trapa quadrispinosa Roxb. hulls extracts on adipogenesis and mitochondrial function in 3T3-L1 adipocytes and C2C12 myotubes and their molecular mechanisms, respectively

許玲瑄(Ling-Hsuan Hsu)

指導教授 ： 徐慶琳

中山醫學大學/健康管理學院/營養學研究所/碩士(2015年)

摘要

肥胖與疲勞為現代人常見的文明病，會造成脂質堆積與肌肉活動力降低。研究指出，植物的果皮與殼含有豐富之多酚化合物，並且具有抗氧化和抗癌等作用。因此，本研究利用不同溶劑 (水、甲醇、乙醇、丙酮和乙酸乙酯) 之四角菱角殼萃取物作為素材，開發成具抗肥胖與抗疲勞功效潛力之保健食品。本研究內容分為三部分：(一) 四角菱角殼不同溶劑萃取、基本組成與抗氧化能力分析。(二) 以細胞模式評估不同溶劑四角菱角殼萃取物之抗肥胖潛力。(三) 以細胞模式評估不同溶劑四角菱角殼萃取物之抗疲勞潛力。本研究結果顯示，(一) 四角菱角殼基本組成以無氮萃取物和粗纖維為主要成分，而四角菱角殼甲醇、乙醇與丙酮萃取物具有較佳的抗氧化能力。(二) 以不同溶劑四角菱角殼萃取物處理3T3-L1脂肪細胞後，篩選出四角菱角殼乙醇萃取物可顯著抑制脂肪細胞胞內三酸甘油酯與甘油-3-磷酸去氫酶活性。在肥胖相關基因表現上，四角菱角殼乙醇萃取物具有提升脂肪細胞分化關鍵基因 (Sirt1) 之表現，進而抑制脂肪細胞分化基因 (PPARγ、C/EBPα和C/EBPβ)、脂肪酸合成基因 (FAS和ACC) 與脂肪細胞激素基因 (Resistin、PAI-1和MCP-1) 之表現。(三) 以不同溶劑四角菱角殼萃取物處理分化後之C2C12肌管細胞，可篩選出四角菱角殼乙醇萃取物具有顯著提升粒線體DNA含量基因 (cytochrome b、COX-I和COX-II) 之表現與粒線體酵素 (citrate synthase和cytochrome c oxidase) 之活性。在抗疲勞相關基因表現上，四角菱角殼乙醇萃取物具有提升粒線體生合成相關基因 (Sirt1、PGC-1α、NRF1、ERRα和TFAM)、粒線體氧化磷酸化相關基因 (NDUFB3、SDHB、COX5b和ATP5 l) 與氧化型肌纖維相關基因 (MyHC I和MyHC IIa) 之表現。綜合上述結果表示，四角菱角殼乙醇萃取物確實具有抑制3T3-L1脂肪細胞脂肪合成與提升C2C12肌管細胞粒線體功能。因此，四角菱角殼乙醇萃取物具有開發為抗肥胖與抗疲勞保健功能材料或產品之潛力。

關鍵字

四角菱角殼萃取物 ； 抗肥胖 ； 3T3-L1脂肪細胞 ； 抗疲勞 ； C2C12肌管細胞

並列摘要

Obesity and fatigue are the most common cases of civilization diseases. They may be accompanied by an increased lipid accumulation and decreased muscle performance, respectively. Previous studies indicated that plant hull and pericarp are rich in polyphenolic compounds which exhibit bioactive functions such as antioxidant and anticancer activities. However, the aim of this study was to investigate the effects of different solvent (water, methanol ethanol, acetone, and ethyl acetate) extracts from Trapa quadrispinosa Roxb. hulls as potential nutraceuticals on in vitro models of anti-obesity and anti-fatigue. There are three topics included in this study: (1) Proximate composition and antioxidant activities of different solvent extracts from Trapa quadrispinosa Roxb. hulls, (2) Effects of different solvent extracts from Trapa quadrispinosa Roxb. hulls on adipogenesis in 3T3-L1 adipocytes, and (3) Effects of different solvent extracts from Trapa quadrispinosa Roxb. hulls on mitochondrial functions in C2C12 myotubes. (1) The proximate composition showed that the nitrogen free extract and crude fiber were the major component of Trapa quadrispinosa Roxb. hulls. The antioxidant activities (total polyphenolics and trolox equivalent antioxidant capacity) in methanol, ethanol, and acetone extracts of Trapa quadrispinosa Roxb. hulls were higher than that in water and ethyl acetone extracts. (2) The result indicated that the ethanol extract of Trapa quadrispinosa Roxb. hulls (EETQRH) significantly decreased the intracellular triglyceride and glycerol-3-phosphate dehydrogenase in 3T3-L1 adipocytes. In gene expressions, EETQRH significantly increased transcription of the key gene (Sirt1), and inhibited the gene levels of adipocytes differentiation (PPARγ, C/EBPα, and C/EBPβ), fatty acid synthesis (FAS and ACC), and adipokines related genes (Resistin, PAI-1, and MCP-1) in 3T3-L1 adipocytes. (3) The result indicated that EETQRH significantly increased the mitochondrial DNA content gene expressions (cytochrome b, COX-I, and COX-II) and enzyme activites (citrate synthase and cytochrome c oxidase) as well. In addition, EETQRH significantly increased the gene levels of mitochondrial functions (Sirt1, PGC-1α, NRF1, ERRα, and TFAM), oxidative phosphorylation (NDUFB3, SDHB, COX5b, and ATP5 l), and oxidative muscle fibers (MyHC I and MyHC IIa) in C2C12 myotubes. These results indicated that EETQRH can suppress adipogenesis in 3T3-L1 adipocytes and promote mitochondrial functions in C2C12 myotubes. Thus, EETQRH may provide a potential ameliorative approach for obesity and fatigue.

並列關鍵字

Trapa quadrispinosa Roxb. hulls extract ； anti-obesity ； 3T3-L1 adipocytes ； anti-fatigue ； C2C12 myotubes

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