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顶端钠-葡萄糖协同转运蛋白 1(SGLT1)的活性和蛋白丰度沿新生仔猪空肠隐窝-绒毛轴表达。

Apical Na+-D-glucose cotransporter 1 (SGLT1) activity and protein abundance are expressed along the jejunal crypt-villus axis in the neonatal pig.

机构信息

Center for Nutrition Modeling, Department of Animal and Poultry Science, University of Guelph, Guelph, Ontario, Canada.

出版信息

Am J Physiol Gastrointest Liver Physiol. 2011 Jan;300(1):G60-70. doi: 10.1152/ajpgi.00208.2010. Epub 2010 Oct 28.

Abstract

Gut apical Na(+)-glucose cotransporter 1 (SGLT1) activity is high at the birth and during suckling, thus contributing substantially to neonatal glucose homeostasis. We hypothesize that neonates possess high SGLT1 maximal activity by expressing apical SGLT1 protein along the intestinal crypt-villus axis via unique control mechanisms. Kinetics of SGLT1 activity in apical membrane vesicles, prepared from epithelial cells sequentially isolated along the jejunal crypt-villus axis from neonatal piglets by the distended intestinal sac method, were measured. High levels of maximal SGLT1 uptake activity were shown to exist along the jejunal crypt-villus axis in the piglets. Real-time RT-PCR analyses showed that SGLT1 mRNA abundance was lower (P < 0.05) by 30-35% in crypt cells than in villus cells. There were no significant differences in SGLT1 protein abundances on the jejunal apical membrane among upper villus, middle villus, and crypt cells, consistent with the immunohistochemical staining pattern. Higher abundances (P < 0.05) of total eukaryotic initiation factor 4E (eIF4E) protein and eIE4E-binding protein 1 γ-isoform in contrast to a lower (P < 0.05) abundance of phosphorylated (Pi) eukaryotic elongation factor 2 (eEF2) protein and the eEF2-Pi to total eEF2 abundance ratio suggest higher global protein translational efficiency in the crypt cells than in the upper villus cells. In conclusion, neonates have high intestinal apical SGLT1 uptake activity by abundantly expressing SGLT1 protein in the epithelia and on the apical membrane along the entire crypt-villus axis in association with enhanced protein translational control mechanisms in the crypt cells.

摘要

肠道顶端钠-葡萄糖共转运蛋白 1(SGLT1)在出生和哺乳期间的活性很高,因此对新生儿的葡萄糖稳态有很大贡献。我们假设新生儿通过在整个隐窝-绒毛轴上表达顶端 SGLT1 蛋白,通过独特的控制机制具有高 SGLT1 最大活性。通过用扩张的肠囊法从新生仔猪的回肠隐窝-绒毛轴上依次分离上皮细胞,制备顶端膜囊泡,并测量其 SGLT1 活性的动力学。结果表明,仔猪的回肠隐窝-绒毛轴上存在高水平的最大 SGLT1 摄取活性。实时 RT-PCR 分析显示,SGLT1 mRNA 丰度在隐窝细胞中比在绒毛细胞中低(P < 0.05)30-35%。在上部绒毛、中部绒毛和隐窝细胞的空肠顶端膜上,SGLT1 蛋白丰度没有显著差异,与免疫组织化学染色模式一致。与磷酸化(Pi)真核延伸因子 2(eEF2)蛋白和 eEF2-Pi 与总 eEF2 丰度的比值相比,总真核起始因子 4E(eIF4E)蛋白和 eIF4E 结合蛋白 1 γ-同工型的丰度更高(P < 0.05)表明隐窝细胞的整体蛋白质翻译效率更高。总之,新生儿通过在整个隐窝-绒毛轴上丰富地表达上皮细胞和顶端膜上的 SGLT1 蛋白,并与隐窝细胞中增强的蛋白质翻译控制机制相关,具有高肠道顶端 SGLT1 摄取活性。

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