基于MKN28/Caco-2串联模型的荞麦多酚差异吸收及其降脂机制

doi:10.3864/j.issn.0578-1752.2026.13.014

摘要/Abstract

摘要：

【背景】多酚类化合物在降脂方面具有显著效果，且其在人体胃肠道中的吸收具有差异性，这一差异性对其生物利用度产生重要影响。已有研究表明，荞麦作为一种富含多酚的天然食品，可能在降脂方面具有独特的作用。【目的】本研究旨在通过建立MKN28/Caco-2连续细胞转运模型，探索荞麦多酚的差异性吸收，结合细胞试验和肠道菌群发酵试验，系统研究荞麦多酚的降脂作用及其机制。【方法】通过超高效液相色谱-四极杆静电场轨道阱质谱（UHPLC-Q-Orbitrap MS）结合数据库分析荞麦多酚提取物。通过建立MKN28/Caco-2连续细胞转运模型，将荞麦多酚分为易吸收和难吸收组分，研究易吸收组分在脂肪细胞中的降脂效应。采用油红O染色法和细胞内脂质测定，评估荞麦多酚的降脂作用。此外，通过体外肠道菌群发酵试验分析难吸收组分对肠道菌群的影响。【结果】荞麦多酚中，鉴定出23种多酚类化合物；其中，芦丁和原儿茶酸等多酚类物质的响应强度较高。油红O染色试验表明，易吸收组分能够减少脂肪细胞中的脂质积累，3T3-L1细胞内的TG、TC和LDL-C积累分别降低了17.8%、28.6%和15.2%。肠道菌群发酵试验显示，难吸收组分能够调节肠道菌群的组成，增加拟杆菌属和双歧杆菌属的相对丰度，并减少志贺氏菌和灵芝菌属的丰度。【结论】荞麦多酚提取物含有23种酚类化合物，易吸收组分具有显著的细胞内降脂效果，可用于肥胖治疗；难吸收组分通过调节肠道菌群的相对丰度发挥代谢调控作用。该研究揭示了荞麦多酚的差异性吸收和双途径作用机制，为荞麦多酚的功能性食品开发提供了理论依据。

关键词: 荞麦多酚, MKN28/Caco-2串联模型, 差异性吸收, 降脂作用, 肠道菌群

Abstract:

【Background】 Polyphenolic compounds exhibit pronounced lipid-lowering effects, and their absorption in the human gastrointestinal tract varies considerably, which critically influences their bioavailability. Buckwheat, a natural food being rich in polyphenols, has been suggested to possess unique lipid-lowering potential. 【Objective】 This study aimed to investigate the differential absorption of buckwheat polyphenols using an MKN28/Caco-2 sequential transport model, and to systematically evaluate their lipid-lowering effects and underlying mechanisms through cellular assays and gut microbiota fermentation experiments.【Method】 Buckwheat polyphenol extract was characterized by ultra-high-performance liquid chromatography coupled with quadrupole Orbitrap mass spectrometry (UHPLC-Q-Orbitrap MS). Based on the MKN28/Caco-2 transport model, the extract was fractionated into readily absorbable and poorly absorbable components. The lipid-lowering effects of the absorbable fraction were assessed in adipocytes using Oil Red O staining and intracellular lipid quantification. In addition, the impact of the non-absorbable fraction on gut microbiota composition was evaluated in vitro fermentation.【Result】 Component analysis revealed that there are 23 kinds of polyphenolic compounds rutin and protocatechuic acid exhibited relatively high signal intensities. The absorbable fraction significantly reduced lipid accumulation in adipocytes, with triglyceride (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C) levels in 3T3-L1 cells decreased by 17.8%, 28.6%, and 15.2%, respectively. Furthermore, in vitro fermentation demonstrated that the poorly absorbable fraction (BWP-AP) modulated gut microbiota composition by increasing the relative abundance of Bacteroides and Bifidobacterium, while decreasing Escherichia-Shigella and Alistipes.【Conclusion】 Buckwheat polyphenol extract contained 23 identified phenolic compounds. The readily absorbable fraction exerted significant intracellular lipid-lowering effects, whereas the poorly absorbable fraction regulated metabolic homeostasis via modulation of gut microbiota. These findings revealed a dual-pathway mechanism involving direct host absorption and microbiota-mediated regulation, providing a theoretical basis for the development of functional foods based on buckwheat polyphenols.

Key words: buckwheat polyphenols, MKN28/Caco-2 continuous transport model, differential absorption, lipid-lowering effects, gut microbiota

姚轶俊, 陈煜, 鞠兴荣, 王立峰. 基于MKN28/Caco-2串联模型的荞麦多酚差异吸收及其降脂机制[J]. 中国农业科学, 2026, 59(13): 2933-2945.

YAO YiJun, CHEN Yu, JU XingRong, WANG LiFeng. Differential Absorption of Components in Buckwheat Polyphenols via MKN28/Caco-2 Continuous Transport Model and Their Lipid- Lowering Effects[J]. Scientia Agricultura Sinica, 2026, 59(13): 2933-2945.

图/表 8

表1

表2

UHPLC-Q-Orbitrap MS对苦荞提取物中多酚组成的鉴定"

序号 Number	保留时间 Retention time	峰面积 Peak area	相对定量 Relative quantification (%)	分子量Molecular weight		误差 Error value	分子式 Formula	注释源Annotation source			化合物 Compounds
序号 Number	保留时间 Retention time	峰面积 Peak area	相对定量 Relative quantification (%)	测量值 Measured value	理论值 Theoretical value	误差 Error value	分子式 Formula	mzCloud	mzVault	MassList	化合物 Compounds
1	5.779	31520316066	95.31302115	610.1529	610.52	0.36715	C₂₇H₃₀O₁₆	√	√	√	芦丁 Rutin
2	5.356	813686384.8	2.460473665	154.027	154.12	0.09301	C₇H₆O₄	√	√	√	原儿茶酸 Protocatechuic acid
3	5.499	173675725.5	0.525171069	756.2135	756.66	0.44649	C₃₃H₄₀O₂₀			√	山茶花苷A Camelliaside A
4	5.478	167558784.7	0.506674297	170.022	170.12	0.09805	C₇H₆O₅	√	√	√	没食子酸 Gallic acid
5	5.685	110062044.7	0.332812208	390.132	390.38	0.24805	C₂₀H₂₂O₈			√	虎杖苷 Polydatin
6	6.032	67686001.22	0.204672988	668.198	668.6	0.40199	C₃₀H₃₆O₁₇			√	丹皮苷B Tenuifoliside B
7	2.392	40018060.9	0.121009011	436.1357	436.41	0.27435	C₂₁H₂₄O₁₀			√	根皮苷 Phlorizin
8	6.302	31067800.42	0.093944677	286.0482	286.236	0.18784	C₁₅H₁₀O₆			√	木犀草素 Luteolin
9	6.179	30683750.47	0.092783364	346.0682	346.29	0.2218	C₁₇H₁₄O₈			√	丁香苷 Syringetin
10	5.696	24698332.19	0.074684297	466.1122	466.39	0.27781	C₂₁H₂₂O₁₂			√	车前草苷 Plantagoside
11	8.147	19850602.56	0.060025442	270.053	270.24	0.18705	C₁₅H₁₀O₅	√	√	√	染料木黄酮 Genistein
12	5.903	16924242.75	0.05117654	194.0584	194.18	0.12162	C₁₀H₁₀O₄	√	√	√	阿魏酸 Ferulic acid
13	9.826	13206999.01	0.039936115	770.2244	770.68	0.45556	C₃₄H₄₂O₂₀			√	香蒲苷 Typhaneoside
14	5.652	12511802.1	0.037833937	304.0952	304.29	0.19482	C₁₆H₁₆O₆			√	原苏木素B Protosappanin B
15	10.915	9320952.514	0.028185255	784.2782	784.75	0.47179	C₃₆H₄₈O₁₉			√	安格洛苷C Angoroside C
16	3.867	5294193.31	0.016008899	304.0581	304.25	0.19188	C₁₅H₁₂O₇			√	二氢槲皮素 Taxifolin
17	6.305	3454208.941	0.010445044	166.0634	166.17	0.10657	C₉H₁₀O₃			√	丹皮酚 Paeonol
18	5.464	3036822.242	0.009182925	492.1045	492.43	0.32549	C₂₆H₂₀O₁₀			√	丹酚酸C Salvianolic acid C
19	6.344	2796773.379	0.008457051	362.174	362.42	0.24605	C₂₀H₂₆O₆			√	开环异落叶松树脂酚 Secoisolariciresinol
20	5.625	1966842.11	0.005947455	272.0901	272.25	0.15994	C₁₂H₁₆O₇			√	熊果苷 Arbutin
21	5.599	1747962.418	0.005285594	582.1941	582.5505	0.35639	C₂₇H₃₄O₁₄			√	柚皮苷 Naringin dihydrochalcone
22	11.716	631804.1539	0.001910487	578.1411	578.52	0.37889	C₃₀H₂₆O₁₂			√	原花青素B2 Procyanidin B2
23	10.054	118564.9798	0.000358524	724.221	724.66	0.43898	C₃₃H₄₀O₁₈			√	女贞苷 Ligustroflavone

表2

图1

图2

图3

图4

图5

图6

参考文献 30

[1]	Sharma T, Agrawal P, Sahoo P, Verma N K, Vasikarla S. Line segments based rotation invariant descriptor for disparate images [C]//2018 IEEE Applied Imagery Pattern Recognition Workshop (AIPR). October 9-11, 2018, Washington, DC, USA. IEEE, 2018: 1-8.
[2]	Xiao J B, Kai G Y, Yamamoto K, Chen X Q. Advance in dietary polyphenols as α-glucosidases inhibitors: A review on structure- activity relationship aspect[J]. Critical Reviews in Food Science and Nutrition, 2013, 53(8): 818-836.
[3]	Kandemir K, Tomas M, McClements D J, Capanoglu E. Recent advances on the improvement of quercetin bioavailability[J]. Trends in Food Science & Technology, 2022, 119: 192-200.
[4]	谢佳函, 刘回民, 刘美宏, 郑明珠, 徐倩, 刘景圣. 杂粮多酚功能活性研究进展[J]. 食品工业科技, 2017, 38(14): 326-329, 335.
	Xie J H, Liu H M, Liu M H, Zheng M Z, Xu Q, Liu J S. Research progress of functional activities of polyphenols in coarse cereals[J]. Science and Technology of Food Industry, 2017, 38(14): 326-329, 335. (in Chinese)
[5]	李辉, 刘志虎, 李华, 卿笃信. 姜黄素脂质体对高脂诱导的非酒精性脂肪性肝病小鼠的降脂保肝作用[J]. 中国医药, 2020, 15(7): 1053-1057.
	Li H, Liu Z H, Li H, Qing D X. Effect of curcumin liposomes on lipid-lowering and hepatoprotective effects of hyperlipidemia-induced nonalcoholic fatty liver disease mice[J]. China Medicine, 2020, 15(7): 1053-1057. (in Chinese)
[6]	徐红敏, 吴惠文. 芦丁抑制3T3-L1前脂肪细胞分化并促进米色脂肪细胞形成[J]. 中国公共卫生, 2021, 37(5): 845-848.
	Xu H M, Wu H W. Rutin inhibits 3T3-L1 preadipocytes differentiation and promotes beige adipogenesis[J]. Chinese Journal of Public Health, 2021, 37(5): 845-848. (in Chinese)
[7]	谢琳琳, 迟晓星, 董佳萍, 杨琪, 王鹤霖, 刘殊凡, 张东杰. 荞麦槲皮素对高脂饮食大鼠脂代谢及脂肪因子的调节作用[J]. 现代食品科技, 2023, 39(5): 1-7.
	Xie L L, Chi X X, Dong J P, Yang Q, Wang H L, Liu S F, Zhang D J. Regulatory effects of buckwheat quercetin on lipid metabolism and adipokines in rats receiving a high-fat diet[J]. Modern Food Science and Technology, 2023, 39(5): 1-7. (in Chinese)
[8]	李洁, 吴金春, 曹萍. 福林酚比色法对含茶制品中茶多酚含量的测定研究[J]. 中国食品工业, 2025(19): 85-87.
	Li J, Wu J C, Cao P. Study on the determination of tea polyphenol content in tea products by Folin phenol colorimetric method[J]. China Food Industry, 2025(19): 85-87. (in Chinese)
[9]	秦博闻. 薏米多酚的吸收转运及其降脂功能研究[D]. 南京: 南京财经大学, 2023.
	Qin B W. Absorption, transport and lipid-lowering function of polyphenols in Coix seed[D]. Nanjing: Nanjing University of Finance & Economics, 2023. (in Chinese)
[10]	张泽宇, 代玉婷, 丁煜哲, 李岩松, 姜珂昕, 张林, 石镜明. 正常肝细胞系LO2中单油酸诱导非酒精性脂肪性肝细胞模型的建立[J]. 中国生物化学与分子生物学报, 2026, 42(1): 143-152.
	Zhang Z Y, Dai Y T, Ding Y Z, Li Y S, Jiang K X, Zhang L, Shi J M. Development of an NAFLD cellular model using monooleic acid in a wild-type liver cell line LO2[J]. Chinese Journal of Biochemistry and Molecular Biology, 2026, 42(1): 143-152. (in Chinese)
[11]	张富新, 贺连珍, 马楠. 荞麦营养成分及生物活性成分研究进展[J]. 北方农业学报, 2025, 53(3): 104-109.
	Zhang F X, He L Z, Ma N. Research progress on nutritional and bioactive components of Fagopyrum esculentum[J]. Journal of Northern Agriculture, 2025, 53(3): 104-109. (in Chinese)
[12]	王丽, 李浩志, 段黎娟, 闫春生, 贾安, 黄小强. 响应面法优化女贞花原儿茶酸提取工艺[J]. 食品工业, 2025, 46(10): 1-4.
	Wang L, Li H Z, Duan L J, Yan C S, Jia A, Huang X Q. Optimization of extraction process of protocatechuic acid from Ligustrum lucidum flowers by response surface methodology[J]. The Food Industry, 2025, 46(10): 1-4. (in Chinese)
[13]	张旭, 陈朝银, 董珺琳, 蔡金燕, 赵声兰. 没食子酸对谷氨酸钠诱导的肥胖小鼠的降脂作用[J]. 中成药, 2017, 39(6): 1115-1119.
	Zhang X, Chen C Y, Dong J L, Cai J Y, Zhao S L. Lipid-lowering effects of Gallic acid on glutamate-induced obese mice[J]. Chinese Traditional Patent Medicine, 2017, 39(6): 1115-1119. (in Chinese)
[14]	Qin B W, Yao Y J, Zhang J, Wang L F. Bioavailability of Coix seed polyphenols in a MKN28/caco-2 continuous transport model and their lipid-lowering effects via modulating adipocyte differentiation of 3T3-L1 cells[J]. Journal of Agricultural and Food Chemistry, 2023, 71(22): 8425-8436.
[15]	左丹, 廖霞, 李瑶, 石芳, 王丽颖, 明建. 基于肠道吸收机制的膳食多酚代谢研究进展[J]. 食品科学, 2017, 38(7): 266-271.
	Zuo D, Liao X, Li Y, Shi F, Wang L Y, Ming J. Progress in research on dietary polyphenols metabolism based on a mechanism involving intestinal absorption[J]. Food Science, 2017, 38(7): 266-271. (in Chinese)
[16]	Kikuchi T, Hayashi A, Ikeda N, Morita O, Tasaki J. Multidrug resistance-associated protein 2 (MRP2) is an efflux transporter of EGCG and its metabolites in the human small intestine[J]. The Journal of Nutritional Biochemistry, 2022, 107: 109071.
[17]	Qu L L, Liu Q Q, Zhang Q, Tuo X X, Fan D D, Deng J J, Yang H X. Kiwifruit seed oil prevents obesity by regulating inflammation, thermogenesis, and gut microbiota in high-fat diet-induced obese C57BL/6 mice[J]. Food and Chemical Toxicology, 2019, 125: 85-94.
[18]	Kim S J, Zaidul I S M, Suzuki T, Mukasa Y, Hashimoto N, Takigawa S, Noda T, Matsuura-Endo C, Yamauchi H. Comparison of phenolic compositions between common and Tartary buckwheat (Fagopyrum) sprouts[J]. Food Chemistry, 2008, 110(4): 814-820.
[19]	Lemmens E, De Brier N, Spiers K M, Ryan C, Garrevoet J, Falkenberg G, Goos P, Smolders E, Delcour J A. The impact of steeping, germination and hydrothermal processing of wheat (Triticum aestivum L.) grains on phytate hydrolysis and the distribution, speciation and bio-accessibility of iron and zinc elements [J]. Food Chemistry, 2018, 264: 367-376.
[20]	de Souza da Costa Brum I, Mafra D, de Souza Gouveia Moreira L, Teixeira K T R, Stockler-Pinto M B, de França Cardozo L F M, Borges N A. Consumption of oils and anthocyanins may positively modulate PPAR-γ expression in chronic noncommunicable diseases: A systematic review[J]. Nutrition Research, 2022, 105: 66-76.
[21]	Liu W, Zhao S Q, Wang J Q, Shi J, Sun Y K, Wang W Q, Ning G, Hong J, Liu R X. Back cover: Grape seed proanthocyanidin extract ameliorates inflammation and adiposity by modulating gut microbiota in high-fat diet mice[J]. Molecular Nutrition & Food Research, 2017, 61(9): 1770096.
[22]	Song Z L, Revelo X, Shao W J, Tian L L, Zeng K J, Lei H, Sun H S, Woo M, Winer D, Jin T R. Dietary curcumin intervention targets mouse white adipose tissue inflammation and brown adipose tissue UCP1 expression[J]. Obesity, 2018, 26(3): 547-558.
[23]	Gao X L, Jia R Z, Xie L, Kuang L H, Feng L, Wan C M. A study of the correlation between obesity and intestinal flora in school-age children[J]. Scientific Reports, 2018, 8: 14511.
[24]	Ferrarese R, Ceresola E R, Preti A, Canducci F. Probiotics, prebiotics and synbiotics for weight loss and metabolic syndrome in the microbiome era[J]. European Review for Medical and Pharmacological Sciences, 2018, 22(21): 7588-7605.
[25]	Bervoets L, Van Hoorenbeeck K, Kortleven I, Van Noten C, Hens N, Vael C, Goossens H, Desager K N, Vankerckhoven V. Differences in gut microbiota composition between obese and lean children: A cross-sectional study[J]. Gut Pathogens, 2013, 5(1): 10.
[26]	Ley R E, Turnbaugh P J, Klein S, Gordon J I. Human gut microbes associated with obesity[J]. Nature, 2006, 444(7122): 1022-1023.
[27]	喻喜华, 王一, 李刘若兰, 杨柳, 冯滢璇, 王鸿超, 肖越. 褐藻多糖对肠道菌群及相关代谢的调节作用[J/OL]. 中国乳品工业.
	YU X H, WANG Y, LI L R L, YANG L, FENG Y X, WANG H C, XIAO Y. Modulatory effects of fucoidan on the gut microbiota and associated metabolism[J/OL]. China Dairy Industry. in Chinese)
[28]	Zeng Q, Li D F, He Y, Li Y H, Yang Z Y, Zhao X L, Liu Y H, Wang Y, Sun J, Feng X, Wang F, Chen J X, Zheng Y J, Yang Y H, Sun X L, Xu X M, Wang D X, Kenney T, Jiang Y Q, Gu H, et al. Discrepant gut microbiota markers for the classification of obesity-related metabolic abnormalities[J]. Scientific Reports, 2019, 9: 13424.
[29]	余雅婷, 赵亮, 郝彦玲, 翟征远. 基于体外发酵模型研究4种植物多酚对肠道菌群的调节作用[J]. 食品工业科技, 2025, 46(23): 190-201.
	Yu Y T, Zhao L, Hao Y L, Zhai Z Y. Modulation of intestinal flora by plant polyphenols based on in vitro fermentation models[J]. Science and Technology of Food Industry, 2025, 46(23): 190-201. (in Chinese)
[30]	Segui-Perez C, Huang L Z X, Paganelli F L, Lievens E, Strijbis K. Probiotic Bifidobacterium bifidum strains desialylate MUC13 and increase intestinal epithelial barrier function[J]. Scientific Reports, 2025, 15: 8778.

	得率 Receiving rate (%)	总酚 Total phenolic content (mg GAE·g^-1)	总黄酮 Total flavonoid content (mg RE·g^-1)
苦荞 Tartary buckwheat	3.25±0.02	20.40±0.29	10.83±0.46
甜荞 Sweet buckwheat	2.50±0.04	13.24±0.54	7.30±0.62