中国食品学报2024,Vol.24Issue(9) :93-104.DOI:10.16429/j.1009-7848.2024.09.009

基于Caco-2细胞模型的辣椒素与槲皮素协同调节小肠糖代谢的分子机制

The Molecular Mechanism of Synergistic Effect of Capsaicin and Quercetin on the Glucose Metabolism in Small Intestine Using Caco-2 Monolayer Model

祝文轩 李彤 王向红 米思
中国食品学报2024,Vol.24Issue(9) :93-104.DOI:10.16429/j.1009-7848.2024.09.009

基于Caco-2细胞模型的辣椒素与槲皮素协同调节小肠糖代谢的分子机制

The Molecular Mechanism of Synergistic Effect of Capsaicin and Quercetin on the Glucose Metabolism in Small Intestine Using Caco-2 Monolayer Model

祝文轩 1李彤 1王向红 1米思1
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作者信息

  • 1. 河北农业大学食品科技学院 河北保定 071000
  • 折叠

摘要

为探究辣椒素与槲皮素协同调节小肠糖代谢的分子机制,借助Caco-2细胞单层模型,研究辣椒素与槲皮素单一及联合处理对Caco-2细胞存活率、细胞膜通透性的影响.采用免疫印迹法测定跨膜蛋白Occludin、胞质蛋白ZO-1、表面生长因子受体EGFR和纤维状肌蛋白F-actin以及葡萄糖转运关键蛋白GLUT2、SGLT1、Na+/K+-ATPase的表达量.结果显示:各试验处理组Caco-2细胞的增殖率均大于75%,表明药物对细胞没有明显毒性,可进行后续试验.Caco-2细胞单层模型的跨膜电阻值为(315.70±26.65)Ω.cm2,加药后各试验组的跨膜电阻值均呈现上升趋势,表明膜通透性变小,细胞屏障功能增强,可延缓小肠对葡萄糖的吸收.同时与空白对照组相比,各试验组上调了跨膜蛋白Occludin(>0.65)、胞质蛋白ZO-1(>0.51)、表面生长因子受体EGFR(>0.19)和纤维状肌蛋白F-actin(>0.04)的相对表达量,并且下调了葡萄糖转运关键蛋白GLUT2(>1.00)、SGLT1(>0.78)、Na+/K+-ATPase(>0.87)的相对表达水平.综合来看,辣椒素与槲皮素在调节小肠糖代谢方面具有协同效应,二者在高剂量水平以1:3比例混合效果最佳,主要通过增强肠道黏膜屏障功能,下调葡萄糖转运关键蛋白水平,从而抑制小肠对葡萄糖的吸收来达到调节糖代谢的效果.

Abstract

The present study was conducted to unravel the molecular mechanism underlying the synergistic effect of cap-saicin and quercetin on the regulation of glucose metabolism in small intestine.The Caco-2 monolayer model was estab-lished to investigate the effect of capsaicin and quercetin,both singly and combined,on the cell viability and cell mem-brane permeability.Additionally,western blotting was utilized to determine the expression levels of Occludin,cytoplasmic protein ZO-1,epidermal growth factor receptor(EGFR)and fibrillar muscle protein F-actin as well as key proteins in-volved in glucose transport including GLUT2,SGLT1 and Na+/K+-ATPase.Results showed that the cell viability of all treatment groups was higher than 75%,demonstrating no toxic effect of capsaicin and quercetin on Caco-2 cells.Admin-istration of capsaicin and quercetin increased the TEER values of Caco-2 monolayer model(315.70±26.65)Ω·cm2,which indicated smaller permeability and thus stronger barrier function.Furthermore,compared to the CK group,cap-saicin and quercetin treatments upregulated the expression levels of Occludin(>0.65),ZO-1(>0.51),EGFR(>0.19),and F-actin(>0.04),but downregulated that of GLUT2(>1.00),SGLT1(>0.78)and Na+/K+-ATPase(>0.87).In sum-mary,capsaicin and quercetin can exert a synergistic effect on the regulation of glucose metabolism in small intestine and high dose levels of capsaicin and quercetin at a ratio of 1∶3 was found to be optimal.The potential molecular mech-anism can be attributed to the enhancement of barrier function of small intestine and downregulation of key proteins in-volved in glucose transport,which consequently hinders glucose absorption to achieve a hypoglycemic effect.

关键词

辣椒素/槲皮素/Caco-2细胞模型/小肠糖代谢/分子机制

Key words

capsaicin/quercetin/Caco-2 cell model/glucose metabolism in small intestine/molecular mechanism

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基金项目

国家自然科学基金青年科学基金项目(32202013)

河北省自然科学基金生物农业联合基金培育项目(C2022204222)

出版年

2024
中国食品学报
中国食品科学技术学会

中国食品学报

CSTPCDCSCD北大核心EI
影响因子:1.079
ISSN:1009-7848
参考文献量6
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