Whistler Center for Carbohydrate Research, 745 Agriculture Mall Drive, Purdue University, West Lafayette, IN 47907-2009, USA ; Department of Food Science, Purdue University, West Lafayette, IN 47907-2009, USA.
Department of Food Science, Purdue University, West Lafayette, IN 47907-2009, USA.
J Clin Biochem Nutr. 2014 Jan;54(1):55-60. doi: 10.3164/jcbn.13-59. Epub 2013 Dec 20.
Using the small intestine enterocyte Caco-2 cell model, sucrase-isomaltase (SI, the mucosal α-glucosidase complex) expression and modification were examined relative to exposure to different mono- and disaccharide glycemic carbohydrates. Caco-2/TC7 cells were grown on porous supports to post-confluence for complete differentiation, and dietary carbohydrate molecules of glucose, sucrose (disaccharide of glucose and fructose), maltose (disaccharide of two glucoses α-1,4 linked), and isomaltose (disaccharide of two glucoses α-1,6 linked) were used to treat the cells. qRT-PCR results showed that all the carbohydrate molecules induced the expression of the SI gene, though maltose (and isomaltose) showed an incremental increase in mRNA levels over time that glucose did not. Western blot analysis of the SI protein revealed that only maltose treatment induced a higher molecular weight band (Mw ~245 kDa), also at higher expression level, suggesting post-translational processing of SI, and more importantly a sensing of maltose. Further work is warranted regarding this putative sensing response as a potential control point for starch digestion and glucose generation in the small intestine.
采用小肠肠上皮细胞 Caco-2 细胞模型,研究了蔗糖酶-异麦芽糖酶(SI,黏膜 α-葡萄糖苷酶复合物)的表达和修饰与不同单糖和双糖血糖碳水化合物暴露的关系。Caco-2/TC7 细胞在多孔载体上生长至汇合后进行完全分化,并使用葡萄糖、蔗糖(葡萄糖和果糖的二糖)、麦芽糖(两个葡萄糖α-1,4 连接的二糖)和异麦芽糖(两个葡萄糖α-1,6 连接的二糖)等饮食碳水化合物分子处理细胞。qRT-PCR 结果表明,所有碳水化合物分子都诱导了 SI 基因的表达,尽管麦芽糖(和异麦芽糖)的 mRNA 水平随时间呈递增趋势,而葡萄糖则没有。SI 蛋白的 Western blot 分析表明,只有麦芽糖处理诱导出更高分子量的条带(Mw~245 kDa),表达水平也更高,提示 SI 的翻译后加工,更重要的是对麦芽糖的感知。需要进一步研究这种假定的感应反应作为小肠淀粉消化和葡萄糖生成的潜在控制点。