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中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (21): 6-11.

所属专题： 畜牧兽医

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

不同浓度赖氨酸对猪IEC碱性氨基酸转运载体mRNA表达的影响

职爱民 左建军 周响艳 邹仕庚 黄志毅 王小兰 叶慧 代发文 冯定远   

  • 收稿日期:2010-02-24 修回日期:2010-06-02 出版日期:2010-11-05 发布日期:2010-11-05

The Influence of Different Lysine Concentration on the Cationic Amino Acid Transporter mRNA Expression of Porcine IEC

  • Received:2010-02-24 Revised:2010-06-02 Online:2010-11-05 Published:2010-11-05

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摘要：

本研究通过建立猪肠粘膜上皮细胞（Intestinal epithelial cells, IEC）原代培养的方法，研究了赖氨酸对体外培养的猪小肠粘膜上皮细胞碱性氨基酸转运载体mRNA表达的影响。用机械刮取初生仔猪小肠粘膜加胶原酶Ⅱ消化的方法，成功地在体外培养了新生仔猪IEC；分别用浓度为0.5 mM、2 mM、8 mM赖氨酸处理细胞96 h，提取细胞总RNA，采用实时荧光定量PCR方法，检测了碱性氨基酸转运载体在不同处理细胞中的表达丰度。结果表明：高浓度的赖氨酸对CAT-1 mRNA的表达有上调作用，差异极显著(P<0.01)，并且呈剂量依赖效应；不同浓度的赖氨酸对CAT-2 mRNA的表达影响不大，8 mM和2 mM赖氨酸组CAT-2 mRNA的表达丰度均有低于0.5 mM的趋势，但二者之间差异不显著(P>0.05)；不同浓度的赖氨酸对y+LAT1 mRNA表达影响差异不显著(P>0.05)；高浓度的赖氨酸可提高4F2hc mRNA的表达；0.5 mM和2 mM赖氨酸组4F2hc mRNA的表达丰度均低于8 mM，且差异显著(P<0.05)；0.5 mM和2 mM赖氨酸组之间差异不显著(P>0.05)。

关键词: 朱砂根, 朱砂根, RAPD, 遗传多样性, 聚类分析

Abstract:

The influence of different lysine concentration on the cationic amino acid transporter mRNA expression porcineIntestinal epithelial cells (IEC) in vitro was studied by the primary culture of porcine IEC. The primary culture of porcine IEC system was established by mechanical and enzymatic (collagenaseⅡ) methods using small intestines removed from colostrum-deprived newborn piglets. Total RNA was isolated from IEC samples after treatment with 0.5 mM, 2mM and 8mM lysine for 96 hours. The cationic amino acid transporters mRNA expression abundances were determined by real-time Q–PCR. The results showed as follows: High concentrations of lysine have significant up-regulation effect on the CAT-1 mRNA expression and there was an obvious dose-dependent effect (P<0.01). Lysine treatment had no distinct effect on the CAT-2 mRNA expression abundance. CAT-2 mRNA expressions of 8mM and 2mM lysine treatment were lower than that of 0.5mM lysine group, but with no significant difference (P>0.05). Lysine treatment had no significant effect on the y+LAT1 mRNA expression abundance (P>0.05). High concentrations of lysine could enhance 4F2hc mRNA expression. 4F2hc mRNA expression abundances of 0.5 mM and 2mM lysine groups were significantly lower than that of 8 mM lysine group (P<0.05). There was no significant difference between 0.5mM and 2mM group (P>0.05).