分子营养 Molecular Nutrition

降钙素基因相关肽对猪小肠上皮细胞钠离子依赖型Ⅱb磷转运蛋白表达及磷吸收的影响

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  • 湖南农业大学动物科学技术学院, 长沙 410128
苏文芹(1989-),女,湖南岳阳人,硕士研究生,从事营养生理学研究。E-mail:susmile89@163.com

收稿日期: 2014-04-01

  网络出版日期: 2014-09-10

基金资助

国家自然科学基金面上项目“猪小肠NPT-Ⅱb调控蛋白的筛选与鉴定”(31172218)

Effects of Calcitonin Gene-Related Peptide on Sodium Dependent Phosphate Cotransporter Type Ⅱb Expression and Phosphorus Absorption in Porcine Intestinal Epithelial Cells

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  • College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China

Received date: 2014-04-01

  Online published: 2014-09-10

摘要

本试验旨在研究不同浓度降钙素基因相关肽(CGRP)对猪小肠上皮细胞钠离子依赖型Ⅱb磷转运蛋白(NaPi-Ⅱb)表达和磷吸收的影响。选用第3~8代猪小肠上皮细胞进行试验,按单因素试验设计,将试验细胞分为1个对照组和5个CGRP(1×10-11~1×10-7 mmol/L)试验组,每组设6个重复。用4-甲基偶氮唑盐(MTT)法测定细胞生长速度,采用比色法测定CGRP干预24 h后细胞上清液磷含量,用RT-PCR法检测NaPi-Ⅱb mRNA相对表达量,以及蛋白质印迹(Western blot)法检测NaPi-Ⅱb蛋白表达。结果表明:与对照组相比,不同浓度CGRP对细胞生长没有显著影响(P>0.05);各试验组均极显著或显著地促进了磷的吸收和NaPi-Ⅱb mRNA的相对表达量(P<0.01或P<0.05);除1×10-10 mmol/L组外均显著提高了NaPi-Ⅱb蛋白的表达(P<0.05)。研究结果表明,CGRP通过诱导小肠上皮细胞中NaPi-Ⅱb的表达促进细胞磷的吸收,其中以1×10-9、1×10-8 mmol/L浓度的CGRP促进效果较为显著。

本文引用格式

苏文芹, 方热军 . 降钙素基因相关肽对猪小肠上皮细胞钠离子依赖型Ⅱb磷转运蛋白表达及磷吸收的影响[J]. 动物营养学报, 2014 , 26(9) : 2752 -2760 . DOI: 10.3969/j.issn.1006-267x.2014.09.038

Abstract

The aim of present study was to investigate the effects of calcitonin gene-related peptide (CGRP) on sodium dependent phosphate cotransporter type Ⅱb (NaPi-Ⅱb) expression and phosphorus absorption in porcine intestinal epithelial cells (IPEC-1). The experiment was designed by single factor random allotment, the 3 to 8 generation cells was allotted randomly into 1 control group and 5 experimental groups with 5 concentrations of CGRP (1×10-11 to 1×10-7 mmol/L), each group consisted of 6 replicates with 1 well/dish per replicate. The cell growth curve was determined by the methyl thiazolyl tetrazolium (MTT) method. The phosphorus content in cell supernatant after 24 h CGRP treated were determined by colorimetric method. The NaPi-Ⅱb mRNA expression was determined by RT-PCR, the NaPi-Ⅱb protein expression was determined by Western blot. The results showed as follows: compared with the control group, no significant difference was found in the cell growth in all CGRP groups (P>0.05). Compared with the control group, all CGRP groups significantly increased phosphorus absorption and NaPi-Ⅱb mRNA expression (P<0.01 or P<0.05). All CGRP groups significantly induced NaPi-Ⅱb protein abundance (P<0.05) except the 1×10-10 mmol/L group. The results indicate that CGRP can increase the phosphorus absorption by inducing the NaPi-Ⅱb expression in porcine intestinal epithelial cell, and the 1×10-9 and 1×10-8 mmol/L groups have better significantly promoting effects.

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