紅豆萃取物減緩肥胖之功效及相關分子機制探討

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doi:10.6342/NTU201703601

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红豆萃取物减缓肥胖之功效及相关分子机制探讨

Studies on the anti-obesity efficacy of adzuki bean (Vigna angularis) extracts and its molecular mechanisms of action

邓家宜 , 硕士　　导师：潘敏雄　　

繁体中文 DOI： 10.6342/NTU201703601

肥胖；红豆；脂肪细胞新生； C57BL/6 ； Wnt/β-catenin 路径；肠道菌相； Obesity ； Adzuki bean (Vigna angularis) ； Adipogenesis ； C57BL/6 ； Wnt/β-catenin pathway ； Gut microbiota

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摘要 │ 参考文献 (136) │ 国际计量

摘要〈TOP〉

随着饮食和生活型态的改变，全世界肥胖人口逐年攀升，已经成为日益严重的健康问题，肥胖会增加罹患许多慢性疾病的风险，例如：第二型糖尿病、非酒精性脂肪肝、心血管疾病及癌症等，如何预防及延缓肥胖成为近年来研究的重要议题之一。红豆 (Vigna angularis) 富含多酚类、皂苷及膳食纤维，在先前的研究中发现红豆萃取物具有保护心血管、抗高血压及改善胰岛素阻抗等功效。因此本研究使用国产老鹰红豆高雄 9 号，利用四种不同方法萃取，并以 3T3-L1 前脂肪细胞模式评估其对脂肪细胞新生作用的影响。结果显示，热水萃取之红豆萃取物 (HWE) 最能有效抑制脂肪细胞内油滴堆积，故选择 HWE 进行动物实验。动物模式以 50% 高脂饮食诱导 C57BL/6 小鼠肥胖，同时给予不同比例的 HWE，评估 HWE 是否具有减缓肥胖之功效，并额外加入一组同时喂食正常饮食及高剂量 HWE 之组别 (NDHWE)。饲养 13 周後牺牲，结果显示，喂食 1% 及 5% HWE 的组别与 high-fat diet (HFD) 组相比，小鼠之体重、白色脂肪组织重量及体脂率在不影响其摄食量的情况下，皆显着下降并呈现剂量效应。血液中总胆固醇及三酸甘油酯也显着的降低，其中 NDHWE 组三酸甘油酯含量更是显着低於 ND 组，表示 HWE 可能具有降血脂之潜力。由肝脏切片则可观察到喂食 HWE 的组别，其肝脏内脂肪空泡的数量和大小明显减少，显示 HWE 可以减缓高脂饮食造成之小鼠肝脏脂肪堆积。在先前的研究中已证实在 3T3-L1 细胞模式中活化 Wnt/β-catenin 路径可以抑制脂肪细胞新生，HWE 可以透过增加 Wnt/β-catenin 路径中 Wnt10b、Dvl2、β-catenin 及 cyclin D1 蛋白质表现量，同时降低 destruction complex 成员 Axin1 表现量，活化 Wnt/β-catenin 路径，进而减少脂肪细胞新生必须之转录因子 PPARγ 及 C/EBPα 表现量，以抑制脂肪细胞新生。喂食高脂饮食後会提高 Firmicutes/Bacteroidetes 之比例 (F/B ratio)，导致肠道菌相失衡，同时给予 HWE 则可降低 F/B ratio，提升菌种多样性，另外也发现 HWE 可以提升肠道内益生菌 Lactobacillus 及 Akkermansia 比例，降低 Blautia 及 Ruminococcus 比例，使小鼠肠道菌相组成恢复至与 ND 组相似，推测 HWE 可以透过调节小鼠肠道菌相而减缓肥胖。综合上述结果，HWE 具有抑制脂肪细胞内脂质累积之能力，并透过活化 Wnt/β-catenin 路径及改善肠道菌相减缓高脂饮食诱导之肥胖，达到降低体脂、血脂及改善脂肪肝之功效。显示未来 HWE 具有相当潜力被开发成不易形成体脂肪之保健食品，提升国产红豆经济价值。

并列摘要〈TOP〉

Due to rapid changes in our diet and lifestyle, obesity population is rising year by year worldwide and become a serious health problem. Obesity is associated with numerous chronic diseases such as type 2 diabetes, nonalcoholic fatty liver, cardiovascular disease and cancer. Adzuki bean (Vigna angularis), one of Taiwan's important economic crops, are known to include many polyphenols, saponins and dietary fiber. The extracts of adzuki bean were found to have cardiovascular protection, anti-hypertenison and insulin resistance improvement effect. In this study we used four different solvent to extract adzuki bean. Further, we investigated the anti-adipogenic effect of different adzuki bean extracts in 3T3-L1 cell model. The results showed that hot water extract (HWE) group had the most significant anti-adipogenic effect, it could inhibit lipid accumulation in adipocyte. Therefore, we choosed HWE for further animal experiments. Five-week-old C57BL/6 male mice were fed with normal diet (ND), high-fat diet (50% energy from fat, HFD), HFD with 1% or 5% HWE (LHWE, HHWE) and ND with 5% HWE (NDHWE) for 13 weeks. Compared with HFD group, body weight, white adipose tissue weight and body fat ratio were significantly reduced in both HWE-treated groups in dose-dependent manner without affecting their food intake. Serum total cholesterol and triglyceride levels were significantly lower in both HWE-treated groups compared with HFD group. NDHWE group also had lower triglyceride content compared with ND group, it suggested that HWE may has potential hypolipidemic effect. The number and size of fat vacuoles in liver lesions were significantly reduced, indicating that HWE could ameliorate steatosis in HFD-induced mice. In previous studies, it has been demonstrated that activating Wnt/β-catenin pathway can inhibit adipogenesis in 3T3-L1 cell model. The results showed that HWE-treated groups inhibited adipogenesis via increasing Wnt10b, Dvl2, β-catenin and cyclin D1 protein levels and decreasing protein levels of Axin1 (destruction complex member), activated Wnt/β-catenin pathway and reduced adipogenesis associated transcription factors PPARγ and C/EBPα expression. Moreover, our studies firmed that mice fed with HFD increasd Firmicutes/Bacteroidetes ratio (F/B ratio) and caused gut microbiota imbalance. Supplementation of HWE altered the composition of the gut microbiota by decreasing F/B ratio, improving bacterial diversity, increasing intestinal probiotics Lactobacillus and Akkermansia and decreasing Blautia and Ruminococcus compared with HFD-treated group, suggesting that HWE could improve obesity through modulating gut microbiota. Taken together, these results suggested that HWE could inhibited lipid accumulation in 3T3-L1 cells and ameliorated high-fat-diet induced obesity by activating Wnt/β-catenin pathway and regulating gut microbiota. HWE may has potential to be developed into functional food to improve metabolic disorders and enhance the economic value of domestic adzuki bean in Taiwan.

参考文献 ( 136 ) 〈TOP〉

Ahn, J.; Lee, H.; Kim, S.; Park, J.; Ha, T., The anti-obesity effect of quercetin is mediated by the AMPK and MAPK signaling pathways. Biochemical and Biophysical Research Communications 2008, 373, 545-549.
连结：
Ahn, J.; Lee, H.; Kim, S.; Ha, T., Curcumin-induced suppression of adipogenic differentiation is accompanied by activation of Wnt/β-catenin signaling. American Journal of Physiology Cell Physiology 2010, 298, 1510-1516.
连结：
Anhê, F. F.; Roy, D.; Pilon, G.; Dudonné, S.; Matamoros, S.; Varin, T. V.; Garofalo, C.; Moine, Q.; Desjardins, Y.; Levy, E., A polyphenol-rich cranberry extract protects from diet-induced obesity, insulin resistance and intestinal inflammation in association with increased Akkermansia spp. population in the gut microbiota of mice. Gut 2015, 64, 872-883.
连结：
Arango, N. A.; Szotek, P. P.; Manganaro, T. F.; Oliva, E.; Donahoe, P. K.; Teixeira, J., Conditional deletion of β-catenin in the mesenchyme of the developing mouse uterus results in a switch to adipogenesis in the myometrium. Developmental Biology 2005, 288, 276-283.
连结：
Aronsson, L.; Huang, Y.; Parini, P.; Korach-André, M.; Håkansson, J.; Gustafsson, J. Å.; Pettersson, S.; Arulampalam, V.; Rafter, J., Decreased fat storage by Lactobacillus paracasei is associated with increased levels of angiopoietin-like 4 protein. PloS one 2010, 5, e13087.
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