葡萄糖转运载体4、葡萄糖转运载体2在罗非鱼不同组织中的表达及其对注射葡萄糖的响应  

Expression of Glucose Transporter 4 and Glucose Transporter 2 in Different Tissues of Tilapia and Its Response to Glucose Injection

作　　者：刘含亮[1] 王纪亭[1] 万文菊[2] 付佩胜[3] 孙敏敏[1] 王红卫[1] 

机构地区：[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院,泰安271000 [3]山东省淡水水产研究所,济南252000

出　　处：《动物营养学报》2014年第11期3500-3509,共10页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金项目(31472288);山东省自然科学基金项目(Y2008D35);山东省高等学校科技计划项目(J09LC05)

摘　　要：本试验旨在研究葡萄糖转运载体(GLUT)4、CLUT2在罗非鱼不同组织中的表达及其对注射葡萄糖的响应。利用实时荧光PCR的方法从罗非鱼肌肉中克隆得到GLUT4的c DNA片段,其Gen Bank登录号为JN900493,大小为603 bp,编码200个氨基酸。通过实时荧光半定量PCR检测,比较G LUT4和G LUT2在肌肉、心脏和肝脏中的表达差异,结果显示G LUT4在肌肉和心脏中的表达量较高,而在肝脏中GLUT4的表达量则较低;GLUT2在肝脏中的表达量最高,而在肌肉和心脏中则表现出极低的表达量。选取体重约为80 g的罗非鱼150尾,随机分2个组,每组3个重复,每个重复25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。结果显示:1)试验组血浆葡萄糖含量在注射葡萄糖后1 h时达到最高,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后3 h时达到最高,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。2)试验组肌肉中GLUT4 mRNA的相对表达量在注射葡萄糖后3 h开始升高,在12 h时达到最高,此时显著高于对照组(P<0.05);试验组心脏中GLUT4 mRNA的相对表达量在注射葡萄糖后1 h即较对照组显著升高(P<0.05),而后随着时间的推移逐渐降低,在6 h时恢复到正常水平;注射葡萄糖后,肝脏中GLUT2 mRNA的相对表达量没有显著变化(P>0.05)。结果表明,注射葡萄糖后瞬时升高了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,而肌肉中GLUT4 mRNA相对表达量的提高又延迟于胰岛素含量的升高,从而加重了罗非鱼对葡萄糖的代谢负担。This experiment was conducted to determine the expression of glucose transporter（ G LUT） 4 and G LUT2 in different tissues of tilapia（ Oreochromis niloticus） and its response to glucose injection. A partial c DNA of G LUT4 was cloned from muscle of tilapia by real-time PCR method. The partial c DNA of G LUT4（ Gen Bank accession No. was JN900493） was 603 bp,and encoded 200 amino acids. The expression differences of G LUT4 and G LUT2 in muscle,heart and liver were compared by semi-quantitative real-time PCR detection. The results showed that the expression level of G LUT4 in muscle and heart was higher,while that in liver was lower; the expression level of G LUT2 in liver was higher,while that in muscle and heart was very low. A total of 150 tilapia with the body weight about 80 g were randomly allotted to 2 groups with 3 replicates per group and 25 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 infection（ 0 h） and at 1,3,6 and 12 h after infection,respectively. The results showed as follows： 1） the maximum plasma glucose content of experimental group was observed at 1 h after injection which was significantly higher than that of control group（ P〈 0.05）,and then began to decreaseed and returned to normal level at 3 h after injection;the maximum plasma insulin content of experimental group was observed at 3 h after injection which was significantly higher than that of control group（ P〈 0.05）,and then began to decrease and returned to normal level at12 h after injection. 2） The relative expression level of G LUT4 mRNA in muscle of experimental group began to increase at 3 h after injection,and reached the maximum at 12 h after injection which was significantly higher than that of control group（ P〈 0.05）; the relative expression level of G L

关 键 词：罗非鱼 葡萄糖 葡萄糖转运蛋白4 葡萄糖转运蛋白2 

分 类 号：S963[农业科学—水产养殖]