动物营养学报 >

2017 , Vol. 29 >Issue 2: 652 - 662

DOI: https://doi.org/10.3969/j.issn.1006-267x.2017.02.035

研究简报 Short communications

胰岛素受体-1在尼罗罗非鱼不同组织中的表达及其对注射葡萄糖的响应

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  • 1. 山东农业大学动物科技学院, 泰安 271000;
    2. 泰山医学院基础医学部, 泰安 271000
刘含亮(1986-),男,山东东平人,硕士研究生,研究方向为鱼类营养与免疫。E-mail:liuhanliang666@163.com

收稿日期: 2016-07-01

  网络出版日期: 2017-02-17

基金资助

国家自然科学基金项目（31472288）；山东省科技发展计划项目（2014GGH210010）；山东省高等学校科技计划项目（J09LC05）

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Expression of Insulin Receptor-1 in Different Tissues of Nile Tilapia and Its Response to Glucose Injection

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  • 1. College of Animal Science and Technology, Shandong Agricultural University, Taian 271000, China;
    2. Department of Basic Medicine, Taishan Medical College, Taian 271000, China

Received date: 2016-07-01

  Online published: 2017-02-17

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摘要

本试验旨在研究胰岛素受体-1（IR-1）在尼罗罗非鱼不同组织中的表达及其对注射葡萄糖的响应。利用PCR扩增的方法从尼罗罗非鱼肌肉中克隆IR-1的cDNA片段，并通过半定量PCR检测，比较IR-1在肌肉、肝脏和心脏中的表达差异。选取体重约为100 g的尼罗罗非鱼160尾，随机分2组，每组4个重复，每个重复20尾。试验组腹腔注射葡萄糖（每100 g体重30 mg），对照组以相同剂量腹腔注射0.7%的无菌生理盐水。在注射前（0 h）和注射后的1、3、6和12 h分别进行采样，测定血浆葡萄糖和胰岛素含量，并通过实时荧光定量PCR检测IR-1在肌肉、心脏和肝脏中的mRNA相对表达量。结果显示：1）克隆出的IR-1 cDNA片段，其GenBank登陆号为JN967750，大小为1 979 bp，编码548个氨基酸。序列分析发现，尼罗罗非鱼的IR-1与其他物种比较具有很高的保守性，并具有丰富的酪氨酸激酶特征性序列。2）IR-1在尼罗罗非鱼肌肉、心脏和肝脏中均有较高的表达量，其中在肝脏和肌肉中的表达量基本一致，而在心脏中的表达量相对较低。3）试验组血浆葡萄糖含量在注射葡萄糖1 h后达到最高，并显著高于对照组（P<0.05），而后开始下降，3 h后恢复到正常水平；试验组血浆胰岛素含量在葡萄糖注射3 h后达到最高，并显著高于对照组（P<0.05），而后开始下降，12 h后恢复到正常水平。试验组肌肉和肝脏中IR-1 mRNA的相对表达量在注射葡萄糖后6 h时达到最高，显著高于对照组（P<0.05），在12 h时恢复到正常水平；试验组心脏中IR-1 mRNA的相对表达量在注射葡萄糖后的12 h内没有发生显著变化（P>0.05）。结果表明，注射葡萄糖后即刻升高了尼罗罗非鱼的血浆葡萄糖含量，相对于血浆葡萄糖含量的升高，血浆胰岛素含量的升高相对延迟，而肌肉和肝脏中IR-1 mRNA相对表达量的提高又延迟于血浆胰岛素含量的升高，从而加重了尼罗罗非鱼对葡萄糖的代谢负担。

关键词： 尼罗罗非鱼; 葡萄糖; 胰岛素; 胰岛素受体-1

本文引用格式

刘含亮, 王纪亭, 万文菊, 孙敏敏, 孟晓, 徐蒙蒙 . 胰岛素受体-1在尼罗罗非鱼不同组织中的表达及其对注射葡萄糖的响应[J]. 动物营养学报, 2017 , 29(2) : 652 -662 . DOI: 10.3969/j.issn.1006-267x.2017.02.035

Abstract

This experiment was conducted to determine the expression of insulin receptor (IR)-1 in different tissues of Nile tilapia (Oreochromis nilotica) and its response to glucose injection. A partial cDNA of IR-1 was cloned from muscle of Nile tilapia by real-time PCR method. The expression differences of IR-1 in muscle, liver and heart were compared by semi-quantitative PCR detection. A total of 160 Nile tilapia with the body weight about 100 g were randomly allotted to 2 groups with 4 replicates per group and 20 fish per replicate. In experimental group, glucose with the dose of 30 mg per 100 g body weight was injected intraperitoneally into the fish, while the fish in control group were injected with 0.7% sterile saline water as the same dose and method. Samples were collected and measured before injection (0 h) and at 1, 3, 6 and 12 h after injection, respectively. The plasma glucose and insulin contents were analyzed, and the relative expression level of IR-1 mRNA in muscle, heart and liver was measured by quantitative real-time PCR detection. The results show as follows:1)the cloned partial cDNA of IR-1 (GenBank accession No. was JN967750) was 1 979 bp, and encoded 548 amino acids. Sequence analysis showed that IR-1 of Nile tilapia had high conservation compared with other species, and had a rich tyrosine kinase characteristic sequence. 2) The higher expression level of IR-1 in muscle, heart and liver was found, the IR-1 expression level in liver and muscle was consistent, while that in heart was relatively lower. 3) The maximum plasma glucose content of experimental group was observed at 1 h after glucose injection which was significantly higher than that of control group (P<0.05), and then began to decrease and returned to normal level at 3 h after injection; the maximum plasma insulin content of experimental group was observed at 3 h after glucose injection which was significantly higher than that of control group (P<0.05), and then began to decrease and returned to normal level at 12 h after injection. The relative expression level of IR-1 mRNA in muscle and liver at 6 h after glucose injection of experimental group was significantly higher than that of control group (P<0.05), and then to return to normal level at 12 h after injection; the relative expression level of IR-1 mRNA in heart had no significant change after glucose injection of experimental group (P>0.05). The results suggest that injecting glucose can instantaneously increase the plasma glucose content of Nile tilapia, but the increase of plasma insulin content is relatively delayed compare with plasma glucose content, and the relative expression level of IR-1 in muscle and liver show a delayed increase relative to plasma insulin content, then the burden of glucose metabolism in Nile tilapia is increased.

Key words： Nile tilapia (Oreochromis nilotica); glucose; insulin; insulin receptor-1

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