生长激素受体基因突变对西藏藏猪生长迟缓的影响

doi:10.11843/j.issn.0366-6964.2016.05.004

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (5): 882-887.doi: 10.11843/j.issn.0366-6964.2016.05.004

生长激素受体基因突变对西藏藏猪生长迟缓的影响

岳敏^#，田雨光^#，万斌，庞炜，吴清洪^*，王玉珏^*

(南方医科大学实验动物中心，广州 510515)

收稿日期:2015-10-12 出版日期:2016-05-23 发布日期:2016-05-21
通讯作者: 吴清洪，高级实验师，主要从事实验动物学研究， Tel：020-61648424，E-mail:wuqhvip@126.com；王玉珏，医学博士，副教授，主要从事人类疾病动物模型制作研究，Tel：020-62789158，E-mail:75848808@qq.com
作者简介:岳敏（1982-），医学博士，实验师，主要从事实验动物分子遗传学研究和人类疾病动物模型制作研究，Tel：020-62789009，E-mail:343779791@qq.com；田雨光（1987-），博士生，主要从事比较医学研究，Tel：020-62789009，E-mail:yuguang.tian@qq.com。岳敏和田雨光为共同第一作者
基金资助:
国家自然科学基金（801402625）；中国博士后自然科学基金（2014M550439）；广东省自然科学基金（S2013010014720）；广东省科技计划项目（2011B060300014）；南方医科大学2015科研助手项目（C1032246）

The Impact of GHR Mutation on Suppressing the Growth in Tibet Mini-pig

YUE Min^#，TIAN Yu-guang^#，WAN Bin，PANG Wei，WU Qing-hong^*，WANG Yu-jue^*

（Laboratory Animal Center，Southern Medical University，Guangzhou 510515，China）

Received:2015-10-12 Online:2016-05-23 Published:2016-05-21

摘要/Abstract

摘要：

本研究旨在探索西藏藏猪生长迟缓成因和分子机理。采用逆转录聚合酶链反应（RT-PCR）技术对西藏藏猪生长激素受体（GHR）基因克隆，采用生物信息学方法分析西藏藏猪生长激素受体基因与其他常见品种猪基因与蛋白序列差异，并构建GHR基因的真核过表达质粒，转染到西藏藏猪胚胎成纤维细胞（PEFs）上进行瞬时表达，通过MTT检测细胞增殖活性，利用实时荧光定量PCR技术检测细胞的胰岛素样生长因子-1（IGF-1）表达的情况。西藏藏猪GHR基因编码区1 716 bp，编码572个氨基酸；与普通猪GHR基因序列进行比对分析表明，在西藏藏猪GHR基因编码区1 225 bp处发生T→G突变，导致丝氨酸突变为丙氨酸；GHR基因真核过表达质粒成功转染PEFs后，PEFs生长活性显著提高；qPCR检测表明，细胞株中IGF-1表达显著上调。综上表明，GHR基因的表达可以提高PEFs细胞的增殖活性，诱导IGF-1基因的表达，GHR基因在1 225 bp处的点突变可能影响西藏藏猪的生长迟缓。

关键词: 西藏藏猪, GHR, IGF-1, 胚胎成纤维细胞

Abstract:

This study aimed to explore the genetic and molecular mechanism of Tibet mini-pig growth retardation.RT-PCR technique was used to amplify Tibet mini-pig GHR gene.Bioinformatics method was used to analyze the differences of GHR gene and protein sequence between Tibet mini-pig and other common variety pigs.The eukaryotic expression vector of common pig GHR gene was transfected into Tibet mini-pig’s embryonic fibroblasts.The MTT assay was used to detect the cell proliferation activity after transfected GHR-pIRES2-EGFP vector.The real-time PCR was used to detect the mRNA expression of the IGF-1 gene.There were 1 716 bp in Tibet mini-pig GHR gene coding region，and 572 amino acids were encoded.Compared with common meat pig GHR gene sequence，the base T in Tibet mini-pig GHR gene coding region 1 225 bp mutated into G，and then serine was replaced by alanine.The growth activity of Tibet mini-pig’s embryonic fibroblasts were significantly improved after transfected with GHR-pIRES2-EGFP，and the mRNA level of IGF-1 was up-regulated significantly.The results showed that GHR could improve the activity of PEFs cell proliferation，and induce the expression of IGF-1 gene；The mutation of GHR gene in 1 225 bp might involve in the process of growth retardation in Tibet mini-pig.

Key words: Tibet mini-pig, GHR, IGF-1, embryonic fibroblasts

中图分类号:

S828.2

岳敏，田雨光，万斌，庞炜，吴清洪，王玉珏. 生长激素受体基因突变对西藏藏猪生长迟缓的影响[J]. 畜牧兽医学报, 2016, 47(5): 882-887.

YUE Min，TIAN Yu-guang，WAN Bin，PANG Wei，WU Qing-hong，WANG Yu-jue. The Impact of GHR Mutation on Suppressing the Growth in Tibet Mini-pig[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(5): 882-887.

参考文献

［1］CASTELLANO G，AFFUSO F，CONZA P D，et al.The GH/IGF-1 axis and heart failure［J］.Curr Cardionl Rev，2009，5(3)：203-215.
［2］BERNASCONI S，BERGOMI A，MILIOLI S，et al.Genetics and molecular biology of the GHRH-GH-IGF-I axis［J］.Minerva Pediatr，1999，51(11-12)：375-394.
［3］同海妮，白淑，王树迎，等.GHR和IGF-IR在济宁青山羊子宫内分布和表达的发育性变化［J］.畜牧兽医学报，2013，44(9)：1392-1399.
TONG H N，BAI S，WANG S Y，et al.Developmental changes of the distribution of GHR and IGF-IR and the expression of its mRNA in the uterus of Jining Grey goats［J］.Acta Veterinaria et Zootechnica Sinica，2013，44(9)：1392-1399.(in Chinese)
［4］王淑燕，霍金龙，潘伟荣，等.版纳微型猪近交系GHR基因克隆及生物信息学分析［J］.中国实验动物学报，2014，22(1)：48-56.
WANG S Y，HUO J L，PAN W R，et al.Cloning and bioinformatics analysis of GHR gene in Banna minipig inbred line［J］.Acta Laboratorium Animalis Scientia Sinica，2014，22(1)：48-56.(in Chinese)
［5］MULLIS P E.Genetics of GHRH，GHRH-receptor，GH and GH-receptor：its impact on pharmacogenetics［J］.Best Pract Res Clin Endocrinol Metab，2011，25(1)：25-41.
［6］RIEDL S，KLUGE M，SCHWEITZER K，et al.Adaptation of ghrelin and the GH/IGF axis to high altitude［J］. Eur J Endocrinol，2012，166(6)：969-976.
［7］FUSCO A，MIELE L，D'UONNOLO A，et al.Nonalcoholic fatty liver disease is associated with increased GHBP and reduced GH/IGF-I levels［J］. Clin Endocrinol (Oxf)，2012，77(4)：531-536.
［8］MILES H L，DERRAIK J G，CHIAVAROLI V，et al.Response to IGF-1 generation test in short prepubertal children born very preterm or at term［J］.Horm Res Paediatr，2015，84(5)：298-304.
［9］FRYSTYK J，DJURHUUS C B，JOHANSEN T，et al.Measurement of free GH and bioactive IGF-I in non-diabetic haemodialysis patients treated with GH for 7 days［J］.Nephrol Dial Transplant，2012，27(11)：4211-4218.
［10］AISENBERG J，AUYEUNG V，PEDRO H F，et al.Atypical GH insensitivity syndrome and severe insulin-like growth factor-I deficiency resulting from compound heterozygous mutations of the GH receptor，including a novel frameshift mutation affecting the intracellular domain［J］.Horm Res Paediatr，2010，74(6)：406-411.
［11］岳敏，范沛，吴丽红，等.广州地区三种小型猪血液生理生化指标的比较［J］.中国比较医学杂志，2011，21(8)：24-26.
YUE M，FAN P，WU L H，et al.Comparison of blood physiological and biochemical parameters among three types of minipigs in Guangzhou［J］.Chinese Journal of Comparative Medicine， 2011，21(8)：24-26.(in Chinese)
［12］TIAN Y G，YUE M，GU Y，et al.Single-nucleotide polymorphism analysis of GH，GHR，and IGF-1 genes in minipigs［J］.Braz J Med Biol Res，2014，47(9)：753-758.
［13］于意，王伟，王莹，等.生长激素受体基因多态性与特发性矮小遗传易感性的关系［J］.上海交通大学学报 (医学版)，2011，31(7)：932-936.
YU Y，WANG W，WANG Y，et al.Associations of single nucleotide polymorphisms of growth hormone receptor with susceptibility to idiopathic short stature［J］.Journal of Shanghai Jiaotong University (Medical Science)，2011，31(7)：932-936.(in Chinese)
［14］LAN H，LI W，JIANG H，et al.Intracellular signaling transduction pathways triggered by a well-known anti-GHR monoclonal antibody，Mab263，in vitro and in vivo［J］.Int J Mol Sci，2014，15(11)：20538-20554.
［15］GAN Y，PATERSON A J，ZHANG Y，et al.Functional collaboration of insulin-like growth factor-1 receptor (IGF-1R)，but not insulin receptor (IR)，with acute GH signaling in mouse calvarial cells［J］.Endocrinology，2014，155(3)：1000-1009.
［16］MA F，WEI Z，SHI C，et al.Signaling cross talk between growth hormone (GH) and insulin-like growth factor-I (IGF-I) in pancreatic islet β-cells［J］.Mol Endocrinol，2011，25(12)：2119-2133.
［17］邓景致.版纳微型猪近交系生长相关基因遗传背景分析［D］.吉林：吉林大学，2012.
DENG J Z.Analysis of genetic background of the growth related genes in Banna Miniature inbreeding pig［D］.Jilin：Jilin University，2012.(in Chinese)

[1]	岳敏，田雨光，万斌，庞炜，吴清洪，王玉珏. 生长激素受体基因突变对西藏藏猪生长迟缓的影响[J]. 畜牧兽医学报, 2016, 47(5): 882-887.
[2]	尹志红，王恩峰，白瑞，庞全海. RNAi对小鼠胚胎成纤维细胞中MC1R基因表达的影响[J]. 畜牧兽医学报, 2014, 45(2): 218-224.
[3]	尹志红，王恩峰，白瑞，庞全海. RNAi对小鼠胚胎成纤维细胞中MC1R基因表达的影响[J]. 畜牧兽医学报, 2014, 45(2): 218-224.

生长激素受体基因突变对西藏藏猪生长迟缓的影响

The Impact of GHR Mutation on Suppressing the Growth in Tibet Mini-pig

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 3

编辑推荐

Metrics

本文评价