中国农业科学 ›› 2023, Vol. 56 ›› Issue (5): 981-994.doi: 10.3864/j.issn.0578-1752.2023.05.013

• 食品科学与工程 • 上一篇    下一篇

荞麦多酚干预对高脂膳食诱导小鼠的降脂作用及其调控机制

姚轶俊(), 鞠兴荣, 王立峰()   

  1. 南京财经大学食品科学与工程学院/江苏省现代粮食流通与安全协同创新中心,南京 210023
  • 收稿日期:2022-04-26 接受日期:2022-11-25 出版日期:2023-03-01 发布日期:2023-03-13
  • 通信作者: 王立峰,E-mail:wanglifeng_8@nufe.edu.cn
  • 联系方式: 姚轶俊,E-mail:yyj@nufe.edu.cn。
  • 基金资助:
    国家重点研发计划(2021YFD2100904); 国家自然科学基金面上项目(32172147); 江苏省现代农业产业技术体系建设项目(JATS [2021]522); 江苏省现代农业重点项目(BE2022317)

Lipid-Lowering Effects and Its Regulation Mechanism of Buckwheat Polyphenols in High-Fat Diet-Induced Obese Mice

YAO YiJun(), JU XingRong, WANG LiFeng()   

  1. College of Food Science and Engineering, Nanjing University of Finance and Economics/Collaborative Innovation Center for Modern Grain Circulation and Safety, Nanjing 210023
  • Received:2022-04-26 Accepted:2022-11-25 Published:2023-03-01 Online:2023-03-13

摘要:

【背景】目前已有相关动物实验证实荞麦对于高脂膳食引起的肥胖和脂代谢紊乱具有良好的干预作用。同时研究表明,棕色脂肪适应性产热可以有效改善人体能量新陈代谢。因此,增加棕色脂肪活性、促进白色脂肪棕色化可作为一种预防肥胖和改善能量代谢疾病的有效途径。【目的】采用原粮及从荞麦中鉴定出的4种主要酚类物质按在原粮中的含量比例配制成多酚干预物进行干预,研究荞麦多酚通过调节白色脂肪棕色化对高脂膳食小鼠的干预及改善作用,系统研究荞麦中发挥降脂功能的主要酚类物质及其对脂代谢的调控机制。【方法】通过UHPLC-Q-Orbitrap质谱结合数据库对荞麦多酚提取物中的酚类物质进行鉴定,并通过相对定量将其中含量最多的主要多酚按其在荞麦原粮中的含量比例配制成多酚复合物。通过添加荞麦饲料以及荞麦多酚复合物干预C57BL/6J小鼠饮食,研究荞麦酚类物质改善脂质代谢的作用。以10%、20%、40%比例的荞麦替换基础饲料和2.5 mg∙mL-1荞麦多酚复配物对小鼠进行饮食干预,探究荞麦对小鼠体重、脏器指数和血脂水平的调节作用。并通过Western-blot和q-PCR探究荞麦饲料及多酚干预对小鼠皮下脂肪组织中与产热相关的蛋白及基因表达的影响。【结果】通过UHPLC-Q-Orbitrap质谱鉴定出荞麦多酚提取物中的20种酚类化合物,其中含量最高的4种酚类化合物羟基肉桂酸、槲皮素、滨蒿内酯和芦丁含量达到酚类总量的80%以上,为荞麦中的主要酚类物质。在饮食干预39 d后,与高脂饮食组相比,添加荞麦饲料以及荞麦多酚干预对小鼠的体重、肝脏指数以及血清中甘油三酯、总胆固醇、低密度脂蛋白、谷草转氨酶和谷丙转氨酶的增加有显著的抑制作用,并且随着荞麦添加量的提高,其抑制作用也增加。同时,添加荞麦饲料以及荞麦多酚干预对小鼠肾脏指数、高密度脂蛋白的降低起到了保护和改善的作用。高剂量40%荞麦和多酚干预组的效果最好,表明荞麦饮食干预对高脂膳食诱导的肥胖小鼠脂代谢异常的改善是其中的酚类物质发挥了主要作用。与正常饮食组相比,高脂饮食组中与棕色脂肪相关的UCP1、PRDM-16和PGC-1α蛋白表达水平显著降低,而与白色脂肪特异性表达的Tcf21和HOXC8水平显著上调(P<0.05)。饮食干预后,荞麦干预组中UCP1、PRDM-16和PGC-1α的蛋白表达水平均显著上调,Tcf21和HOXC8的蛋白表达均显著下降(P<0.05)。此外,40%荞麦和多酚干预组还有效地提高了小鼠皮下脂肪组织中UCP1、PRDM-16PGC-1α的表达(P<0.05),并下调Tcf21HOXC8的表达。【结论】荞麦干预可以有效改善高脂饲料诱导小鼠脂代谢紊乱情况,并证实了荞麦多酚在其中发挥了主要作用,其作用机理主要是荞麦多酚可以显著提高棕色脂肪产热活性,同时下调白色脂肪特异性表达,调节能量代谢平衡,改善高脂饮食诱导的高脂血症和能量代谢紊乱。研究结果为荞麦的利用和相关降脂功能食品的开发提供了新的研发思路和理论依据。

关键词: 荞麦, 多酚, 高脂膳食小鼠, 降脂作用, 白色脂肪棕色化

Abstract:

【Background】At present, relevant animal experiments have confirmed that buckwheat had a good intervention effect on obesity and lipid metabolism disorder caused by high-fat diet. At the same time, studies have shown that the adaptive thermogenesis of brown fat could effectively improve the body’s energy metabolism. Therefore, increasing the activity of brown fat and promoting the browning of white fat could be used as an effective way to prevent obesity and improve energy metabolism diseases.【Objective】The raw buckwheat and four kinds of main polyphenols identified from buckwheat were prepared according to the content proportion to study the lipid-lowering effects of buckwheat polyphenols in high-fat diet-induced obese mice via browning of white adipocytes.【Method】Phenolic compounds in buckwheat polyphenol extract were identified by UHPLC-Q-Orbitrap mass spectrometry combined with database. And the main polyphenol compound was prepared according to the content proportion in the raw buckwheat. In order to explore the regulating effects of buckwheat on body weight, organ index and blood lipid level in C57BL/6J mice, the dietary intervention was carried out with 10%, 20% and 40% proportion of buckwheat to replace basic feed and the 2.5 mg∙mL-1 of mixture buckwheat polyphenol standard substance. Last but not least, Western-blot and q-PCR were used to investigate the effects of buckwheat feed and polyphenol intervention on the expression of heat-producing proteins and genes in white and brown fat in mouse subcutaneous adipose tissue.【Result】Twenty phenolic compounds in buckwheat polyphenol extract were identified by UHPLC-Q-Orbitrap mass spectrometry, and the content of four phenolic compounds with the highest content (hydroxycinnamic acid, quercetin,scoparone, and rutin) accounted for more than 80% of the total phenolic compounds, which were the main polyphenols in buckwheat. The results showed that after 39 d in the diet intervention, compared with high-fat diet group, adding buckwheat feed and buckwheat polyphenol intervention on body weight in mice, the liver index and serum triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-c), aspartate aminotransferase (ALT) and third transaminase (AST) increased significantly inhibitory effect; with the increase of amount of buckwheat, the inhibition increased. At the same time, adding buckwheat feed and buckwheat polyphenol could protect and improve the decrease of renal index and HDL-c in mice. It was also the high dose 40% buckwheat and polyphenol group that had the best effect, which showed that phenolic substances played a major role in the improvement of abnormal lipid metabolism in high-fat diet-induced obese mice. UCP1, PRDM-16, and PGC-1α protein and gene expression levels were significantly decreased in the high-fat diet group compared with the normal diet group, while Tcf21 and HOXC8 expression levels were significantly increased (P<0.05). After dietary intervention, protein expression levels of UCP1, PRDM-16 and PGC-1α in the buckwheat intervention group were significantly up-regulated, while protein expression levels of Tcf21 and HOXC8 were significantly decreased (P<0.05). In addition, the 40% buckwheat and polyphenol intervention group significantly increased the expression of UCP1, PRDM-16 and PGC-1α genes in mouse visceral adipose tissue (P<0.05), and which down-regulated the expressions of Tcf21 and HOXC8 genes.【Conclusion】Buckwheat intervention could effectively improve the lipid metabolism disorder in high-fat diet-induced obese mice, and it was confirmed that buckwheat polyphenols played a major role. The mechanism was mainly buckwheat polyphenols could significantly improve the thermogenic activity of brown fat, reduce the content of white fat, and regulate the energy metabolism balance. This study provided a new idea and theoretical basis for the utilization of buckwheat and the development of functional foods about lipid-lowering products.

Key words: buckwheat, polyphenols, high-fat diet-induced mice, lipid-lowering effects, browning of white adipocytes