以細胞及動物模式探討梅精複方在抗肥胖之作用與分子機制

肥胖是一個全球化的議題，對人們的健康帶來許多的負面影響，同時亦是造成多種疾病的危險因子。近年來，科學家積極尋找各種可以預防或改善肥胖的方法。梅精 (Mei-Gin) 是梅果實的濃縮萃取物，研究指出，其具降低血中三酸甘油酯與血糖之作用。但由於梅精在製作過程中需經長時間的加熱，造成梅果實中所含有之機能性成分被破壞，因而降低其生理功效與價值性。本實驗將利用複方結合方式，以梅精為基礎外加不同比例之梅果萃取物、黑蒜萃取物和仙草萃取物之梅精複方，探討不易形成體脂肪之梅精複方保健食品開發。本實驗分為三個部分，(一) 以體外3T3-L1細胞模式評估梅精複方之抗肥胖之潛力，從7種梅精複方中篩選出具有抗肥胖潛力之樣品；(二) 以體內動物模式評估具潛力梅精複方之抗肥胖潛力 (動物預實驗)，以6週齡的Wistar雄性大鼠作為實驗動物，以高脂飲食誘導之肥胖模式，並給予不同劑量之具潛力梅精複方 (100和300 mg/kg rat)，篩選出最具有抗肥胖潛力之梅精複方；(三) 以體內動物模式評估最具潛力梅精複方之抗肥胖作用與分子機制探討 (動物主實驗)，以6週齡的Wistar雄性大鼠作為實驗動物，以高脂飲食誘導之肥胖模式，並給予不同劑量的最具潛力梅精複方 (50、100和300 mg/kg rat)，評估梅精複方抗肥胖之作用。結果顯示，(一) 在7種梅精複方中，以梅精複方-3和-7在抑制3T3-L1脂肪細胞胞內三酸甘油酯含量與甘油-3-磷酸去氫酶活性之效果最佳，為最具有抑制脂質生合成潛力之樣品；(二) 由動物預實驗結果得知，梅精複方-7較梅精複方-3具有較佳降低體重、肝臟重量和脂肪組織重量之作用。因此，後續研究將挑選梅精複方-7作為第三部分動物主實驗之樣品；(三) 由動物主實驗結果得知，梅精複方-7 可降低高脂飲食誘導之肥胖大鼠之體重、肝臟重量，脂肪組織重量及肝臟總脂質，並增加糞便中脂質排出，在肝臟及腎周圍脂肪組織切片上亦有相同之結果。在梅精複方-7抗肥胖作用機轉上，由脂肪組織基因表現結果得知，給予梅精複方-7可提升高脂飲食誘導肥胖大鼠之Sirt1、ATGL、FOXO1、HSL、ACO、CPT-1、PGC-1 beta、adiponectin和UCP1基因表現，並抑制其FAS、FATP-1和perilipin之基因表現。此外，在肝臟基因表現上，給予梅精複方-7可提升高脂飲食誘導肥胖大鼠之Sirt1、ATGL、FOXO1、HSL、ACO、CPT-1、PGC-1 beta、PPAR-alpha和AMPK之基因表現。在棕色脂肪基因表現上，給予梅精複方-7可提高C/EBPβ、PPAR-gamma、PRDM16、UCP-1、BMP7、PGC-1 alpha、AMPK和Ebf2之基因表現，以及抑制ACC之基因表現。綜合以上的結果顯示，梅精複方-7可作為具抗肥胖潛力之保健食品開發參考。

Obesity has become a known globalized issue. Accompanied with many negative effects on the human body, it is also a major contributing factor for many diseases. In recent years, scientists are constantly searching for alternative to prevent or reduce obesity. Mei-Gin is a concentrated extract of plum fruit which is proven to reduce blood triglyceride and blood sugar. However, a known issue of the functional components contained in the plum fruit could be destroyed during production at the long heated procedure. This experiment will be using combinations of Mei-Gin with different proportions of plum fruit extract, black garlic extract and hsian-tsao extract to recombine a new Mei-Gin formula and to investigate the anti-obesity effect of the Mei-Gin Formula. The experiment is divided into three parts: (1) Assess the anti-obesity potential of Mei-Gin formulas by using the in vitro cell model, and will select the most effective samples among the seven tested Mei-Gin formulas. (2) Evaluate the anti-obesity potential of Mei-Gin formula with the in vivo animal model. By using 6-week-old male Wistar rats as experimental animals, giving high-fat diet to induce obesity and are treated orally with different doses of Mei-Gin Formula-3 and -7 (100 and 300 mg/kg rat) to find most effective formula and it will be involved in the third part of the experiment. (3) Evaluate the anti-obesity effect of Mei-Gin Formula-7 with 6-week-old high-fat diet-induced obese male Wistar rats. The obese rats are treated orally with different doses of Mei-Gin Formula-7 (50, 100, and 300 mg/kg rat) to investigate the anti-obesity effects of Mei-Gin Formula-7. The results have shown that (1) 3T3-L1 cell which treated with Mei-Gin Formula-3 and -7 have significantly decreased in triglyceride and glycerol-3-phosphate dehydrogenase activity. Mei-Gin Formula-3 and 7 are the most effective samples among 7 kinds of Mei-Gin formulas. (2) In animal model, the results indicated that Mei-Gin Formula-7 has better anti-obesity potential compare to Mei-Gin Formula-3. Mei-Gin Formula-7 (100 and 300 mg/kg rat) has significantly reduced the final body weight, liver weight and total body fat of the high-fat diet-induced obese rats. (3) The results have shown that Mei-Gin Formula-7 has significantly reduced the final body weight, liver weight, total body fat, hepatic total lipid and increased the excretion of fecal total lipid. In gene expressions, treated with Mei-Gin Formula-7 upregulated the gene expressions of Sirt1, ATGL, FOXO1, HSL, ACO, CPT-1, PGC-1 beta, adiponectin, and UCP1, whereas it downregulated the gene expression of FAS, FATP-1, perilipin in the perirenal adipose tissue of high-fat diet-induced obese rats. In addition, Mei-Gin Formula-7 also upregulated the gene expression of Sirt1, ATGL, FOXO1, HSL, ACO, CPT-1, PGC-1 beta, PPAR-alpha, and AMPK in the liver of high-fat diet-induced obese rats. Moreover, Mei-Gin Formula-7 could upregulate the gene expression of C/EBPβ, PPAR-gamma, PRDM16, UCP-1, BMP7, PGC-1 alpha, AMPK, and Ebf2, but downregulated ACC gene expression in the brown adipose tissue of high-fat diet-induced obese rats. The results demonstrated the anti-obesity effect of Mei-Gin Formula-7, therefore Mei-Gin Formula-7 could be developed as an anti-obesity potential nutraceutical product.