檸檬皮萃取物對3T3-L1脂肪細胞分化與葡萄糖吸收之影響

Effect of lemon peel extract on 3T3-L1 adipocyte differentiation and glucose uptake

柯永泰(Yung-Tai Ke)

指導教授 ： 吳美莉

國立屏東科技大學/農學院/食品科學系所/碩士(2020年)

https://doi.org/10.6346/NPUST202000345

摘要

過去十年來過氧化物酶體增殖物活化受體-γ（Peroxisome proliferator-activated receptor gamma，PPARγ）已成為研究第二型糖尿病(Type 2 Diabetes Mellitus)藥物標靶之一。透過在脂肪細胞中，PPARγ調控前脂肪細胞分化成脂肪細胞，來影響脂肪細胞的葡萄糖吸收，藉此調節胰島素敏感性，因此活化PPARγ的化合物已成為熱門的研究方向。本實驗探討檸檬皮萃取物是否會活化PPARγ進而影響3T3-L1脂肪細胞分化與葡萄糖吸收。結果顯示檸檬皮萃取物上調脂肪分化相關轉錄因子(PPARγ、CCAAT-enhancer-binding proteins beta和CCAAT-enhancer-binding proteins alpha )及下游脂肪特異性基因 (脂聯素和葡萄糖轉運蛋白4 ) mRNA表現，進而促進前脂肪細胞脂肪累積，並促進其分化為脂肪細胞。接著透過TR-FRET PPARγ Competitive Binding Assay和Dual Luciferase® Reporter Assay分析檸檬皮萃取物對於PPARγ結合性與轉錄活性，結果表示檸檬皮萃取物為PPARγ配體。最後以2-NBDG Glucose Uptake Assay檢測葡萄糖吸收之能力，結果顯示檸檬皮萃取物作用於成熟脂肪細胞時，隨著濃度提高顯著增加葡萄糖吸收。綜合以上結果，檸檬皮萃取物活化PPARγ進而影響脂肪分化與葡萄糖吸收。

關鍵字

過氧化物酶體增殖物活化受體-γ ； 脂肪細胞分化 ； 葡萄糖吸收 ； 第二型糖尿病 ； 檸檬皮萃取物

並列摘要

Over the past decade, peroxisome proliferator-activated receptor-γ（PPARγ）has become one of the most researched targets for anti-Type 2 diabetes mellitus drugs. In adipocytes, PPARγ is a master regulator of the differentiation of preadipocytes into adipocytes, which affects the uptake of glucose by adipocytes to regulate insulin sensitivity. Therefore, compounds that activate PPARγ have become highly sought after in T2DM research. In this experiment, lemon peel extract was used to explore whether it can activate PPARγ and thus affect the differentiation of adipocytes and glucose uptake in 3T3-L1 cells. The results showed that the lemon peel extract up-regulated the mRNA expression of adipocyte differentiation transcription factors (PPARγ, CCAAT-enhancer-binding proteins-β and CCAAT-enhancer-binding proteins-α) and affected adipocyte specific genes ( adiponectin, glucose transporter 4) consequently promoting lipid accumulation of adipocytes. The TR-FRET PPARγ Competitive Binding Assay and Dual Luciferase® Reporter Assay System were used to analyze the transcription activity and binding activity of lemon peel extract for PPARγ. The results indicated that lemon peel extract is a PPARγ ligand and increase transcription activity. Finally, 2-NBDG Glucose Uptake Assay was used to test for glucose uptake in differentiated mature adipocytes. The results showed a significant concentration-dependent increase in glucose uptake. Based on the above results, lemon peel extract may have the potential to stimulate the activation of PPARγ in adipocyte differentiation and increase glucose uptake.

並列關鍵字

Peroxisome proliferator-activated receptor-γ ； Adipocyte Differentiation ； Glucose uptake ； Type 2 diabetes mellitus ； Lemon peel extract

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