Differential proteome analysis along jejunal crypt-villus axis in piglets
Differential proteome analysis along jejunal crypt-villus axis in piglets
复制标题
仔猪空肠隐窝-绒毛轴的差异蛋白质组分析
DOI：
10.2741/4392
复制
发表时间：
2016-01-01
影响因子：
3.1
通讯作者：
Yin, Yulong
中科院分区：
文献类型：
--
作者：
Xiong, Xia;Yang, Huansheng;Yin, Yulong
肠上皮细胞的刷状缘膜负责消化和吸收小肠中的营养物质和离子。为了鉴定与隐窝-绒毛轴上皮细胞成熟相关的BBM蛋白，从21日龄哺乳仔猪的绒毛顶端至空肠隐窝依次分离肠上皮细胞。在制备BBM囊泡后，我们利用相对和绝对定量(ITRAQ)技术，通过等压标记检测到空肠上皮细胞中的194种蛋白质。其中56个BBM蛋白沿隐窝-绒毛轴差异表达。在分化过程中，与营养物质消化吸收相关的蛋白质在绒毛顶端的表达主要是在上、中绒毛或隐窝下调，而与结构和酶调节蛋白相关的蛋白质表达则在很大程度上上调。我们通过蛋白质印迹或免疫化学分析验证了Na+/K+转运ATPase、Galectin-3和肠型脂肪酸结合蛋白的差异。BBM相关蛋白的鉴定有助于加深我们对仔猪和包括人类在内的其他哺乳动物的消化和吸收的了解。
The brush-border membrane (BBM) of enterocytes is responsible for the digestion and absorption of nutrients and ions in the small intestine. To identify the BBM proteins involved in epithelial cell maturation along the crypt-villus axis, enterocytes were sequentially isolated from the villus tip to the crypt of the jejunum from 21-day-old suckling piglets. After preparation of BBM vesicles, we detected 194 proteins in the jejunal epithelial cells by isobaric tags using relative and absolute quantification (iTRAQ) techniques. Of these, 56 BBM proteins were differentially expressed along the crypt-villus axis. During differentiation, the expression of proteins related to digestion and absorption of nutrients was primarily downregulated at the upper, middle villus, or crypt compared to the villus tip, while expression of proteins related to structural and enzyme regulator proteins was largely upregulated. We verified the differences in Na+/K+ -transporting ATPase, galectin-3, and an intestinal-type fatty acid binding protein by western blot or immunochemical analysis. Identification of BBM-associated proteins helps enhance our understanding of digestion and absorption in piglets and other mammals, including humans.