特殊脂肪酸及天然抗氧化物對3T3-L1脂肪細胞生合成影響; The adipogenic effect of uncommon fatty acids and natural antioxidants on 3T3-L1 cell

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题名:	特殊脂肪酸及天然抗氧化物對3T3-L1脂肪細胞生合成影響;The adipogenic effect of uncommon fatty acids and natural antioxidants on 3T3-L1 cell
作者:	賴姿文;Tzu-Wen Lai
贡献者:	中國醫藥大學：營養學系碩士班
关键词:	桐酸;植烷酸;脂肪細胞生合成;α-eleostearic acid;phytanic acid;adipogenesis
日期:	2007-07-23
上传时间:	2009-08-11 15:30:59 (UTC+8)
摘要:	肥胖是全球普遍之健康問題，從天然食物中尋找抗肥胖成分，不論在疾病預防及健康食品市場上均有其重要性。本研究旨在利用3T3-L1前脂肪細胞測試2種特殊脂肪酸及5種天然抗氧化物，對脂肪細胞生合成(adipogenesis)的影響，選擇的二種特殊脂肪酸為α-eleostearic acid (c9,t11,t13 conjugated linolenic acid；c9,t11,t13-CLN)及phytanic acid (PA)，前者存在於苦瓜中，後者在反芻動物肉和乳汁中可發現，5種多酚為EGCG、curcumin、resveratrol、apigenin、genistein存在天然蔬果。分別在3T3-L1 adipogenesis的三個階段﹕前脂肪細胞增生、分化及分化末期階段處理細胞，結果發現脂肪酸都具有促進3T3-L1分化的能力，但由於飽和程度不同，所以促分化能力也會有所不同，分別是SFA<MUFA<PUFA；因此在特殊脂肪酸部分特別選用類似的脂肪酸作為對照。當c9,t11,t13-CLN與其對照(linolenic acid；C18: 3) 相較，會顯著抑制前脂肪細胞增生；藉由細胞內TG (triglyceride)、細胞內G3PDH (glycerol-3-phosphate dehydrogenase) 活性及油紅染色觀察，發現於分化階段c9,t11,t13-CLN會顯著抑制脂肪細胞分化；在分化末期階段會抑制脂質生合成；除了成熟脂肪細胞外c9,t11,t13-CLN於100μM會造成前脂肪細胞及分化階段細胞凋亡，顯示c9,t11,t13-CLN的抑制3T3-L1增生及分化作用與誘導細胞凋亡有關；PA與其對照(palmitic acid；C16:0) 相較，在前脂肪細胞階段，不會造成細胞數目減少；在分化階段則會抑制脂肪細胞分化；在分化末期階段PA會抑制成熟脂肪細胞lipogenesis。多酚類的部分與vehicle control相比較，發現測試的5種多酚類均會抑制增生且呈現dose-response，其中curcumin於50μM會造成細胞毒性；分化階段除了EGCG以外另外4種多酚類都會明顯抑制3T3-L1分化；在成熟脂肪細胞階段，3種多酚類(EGCG、Resveratrol、Genistein)會抑制細胞內TG堆積；除了前脂肪細胞外，curcumin會造成分化階段及分化末期階段細胞凋亡，顯示curcumin抑制3T3-L1分化和抑制成熟脂肪細胞TG堆積與誘導細胞凋亡有關。由以上可知，特殊脂肪酸中c9,t11,t13-CLN與對照組(C18:3)相比，PA與對照組(C16:0)相比，均會抑制脂肪細胞增生、分化、成熟脂肪細胞脂質生合成，而以c9,t11,t13-CLN效果最強；在多酚類當中則是以Curcumin之anti-adipogenesis效果最強，而這些成分具有抗肥胖潛力，值得日後進一步研究。 Obesity is a worldwide health problem. It is important to find the compounds from food with anti-obesity effecct for preventing obesity-related disease and developing functional food. In this study, we investigated the anti-adipogenic effect of two uncommon fatty acids and five natural antioxidants on preadipocyte cell line 3T3-L1. The uncommon fatty acids we chose are α-eleostearic acid (c9,t11,t13 conjugated linolenic acid；c9,t11,t13-CLN) and phytanic acid (PA). c9,t11,t13-CLN is enriched in bitter gourd. Phytanic acid is abundant in ruminant meat and milk. The natural antioxidants we chose are including EGCG, curcumin, resveratrol, apigenin and genistein which are present in fresh vegetables and fruits. These compounds were incubated with 3T3-L1 at three different stages: preadipocyte proliferation, differentiation and terminal differentiation. Results show all common fatty acids could induce the differentiation of 3T3-L1. The potency of inducing adipocyte differentiation was as following, SFA<MUFA<PUFA. During the preadipocyte proliferation stage, c9,t11,t13-CLN reduced the cell number as compared with its control (linolenic acid;C18:3). Compared with C18:3, c9,t11,t13-CLN inhibited 3T3-L1 differentiation as indicated by cellular triglyceride、glycerol-3-phosphate dehydrogenase (G3PDH) activity and oil-red O staining. In terminal differentiation stage, c9,t11,t13-CLN reduced lipogenesis in mature adipocytes. c9,t11,t13-CLN induced apoptosis in the preadipocytes and differentiating adipocytes. Palmitic acid was used as a control group of PA. At preadipocyte proliferation stage, PA didn’t decrease cell number as compared with its control. At differentiation stages, PA suppressed TG accumulation and G3PDH activity in differentiating adipocytes as compared with palmitic acid. The same result were observed by oil-red O staining. At terminal differentiation stages, PA reduced lipogenesis in mature adipocytes. When compared with their vehicle control,five polyphenols reduced the preadipocyte cell number during the proliferation stage, especially for curcumin. At differentiation stage, curcumin, resveratrol, apigenin and genistein resulted in an inhibition of adipocyte differentiation. At terminal differentiation stage, EGCG, resveratrol and genistein suppressed TG accumulation in mature adipocytes. Curcumin induced apoptosis in the differentiating and mature adipocytes. It is concluded that uncommon fatty acids c9,t11,t13-CLN and PA may exert its anti-obesity effect by inhibiting preadipocyte proliferation, differentiation and terminal differentiation. c9,t11,t13-CLN was more potent than PA. Among the polyphenols we tested, curcumin was the most potent in anti-adipogenesis. Thus, these compounds merit further study for their action mechanism and their contribution in developing the health food for anti-obesity.
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