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

作　　者：祝文轩 李彤 王向红[1] 米思 Zhu Wenxuan;Li Tong;Wang Xianghong;Mi Si(College of Food Science and Technology,Hebei Agricultural University,Baoding 071000,Hebei)

机构地区：[1]河北农业大学食品科技学院,河北保定071000

出　　处：《中国食品学报》2024年第9期93-104,共12页Journal of Chinese Institute Of Food Science and Technology

基　　金：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)。

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·cm^(2),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。The present study was conducted to unravel the molecular mechanism underlying the synergistic effect of capsaicin and quercetin on the regulation of glucose metabolism in small intestine.The Caco-2 monolayer model was established to investigate the effect of capsaicin and quercetin,both singly and combined,on the cell viability and cell membrane 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 involved 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.Administration of capsaicin and quercetin increased the TEER values of Caco-2 monolayer model(315.70 ±26.65) Ω·cm~2,which indicated smaller permeability and thus stronger barrier function.Furthermore,compared to the CK group,capsaicin 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 summary,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 mechanism can be attributed to the enhancement of barrier function of small intestine and downregulation of key proteins involved 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 

分 类 号：TS201.4[轻工技术与工程—食品科学]