Abstract：Resistin (RETN) is a specifically produced hormone by fat cells, which plays an important role in glucose and lipid metabolism and also is one of popular candidate genes in animal fat deposition and metabolism. Aiming at study the expression pattern of RETN gene in rump-fat deposition and metabolism of sheep (Ovis aires), long fragment PCR, bioinformatics method and semi-quantitative RT-PCR were used to amplify the full-length sequences, analyze the sequences and detect gene expression profiling of sheep RETN gene, respectively. Fluorescence quantitative Real-time PCR (qRT-PCR) method was used to analyze the expression change of RETN gene between starving and its control groups based on starving model in Altay sheep. The results of cloning and sequence analysis showed that RETN gene encoded 109 amino acids (GenBank No. KJ704841), which was different from the published RETN sequence in GenBank database. RETN mRNA expression profiling showed that RETN expressed in liver, spleen, lung and rump fat differently, especially highly in the liver, significantly higher than that of other groups (P＜0.01), which indicated that RETN gene might play important physiological and biochemical functions in fat synthesis and metabolism in the liver. The result of qRT-PCR showed that RETN mRNA significantly differentially expressed in between starving group and its control groups, with a expression level in persistent starvation group of 5.2 times than that of control group, suggesting that RETN gene might play an important role in fat mobilization. The above findings provide fundamental data for further study on RETN biological function in fat-rump sheep.
[1]甘尚权, 沈敏, 李欢, 等.2013. X染色体60149273位点在脂尾X染色体60149273位点在脂尾（臀）、瘦尾绵羊品种中的多态性及其基因定位研究[J]. 中国农业科学, 46(22): 4791-4799. (Gan SQ, Shen M, Li H, et al.2013. Polymorphism of the 60149273th Loci on X Chromosome Among Fat Tail and Thin Tail Breeds and Its Gene Mapping[J]. Scientia Agriculture Sinica, 46(22): 4791-4799.)
[2]马桢, 郝耿, 杨会国, 等.2012. 阿勒泰羊品种资源现状及发展思路[J]. 草食家畜, 155(02): 10-15.(Ma Z, Hao G, Yang HG, et al.2012. Altay Sheep Status and Development[J]. Grass-Feeding Livestock, 155: 10-15.)
[3]王世银, 张伟, 沈敏, 等. 2013. 绵羊X染色体59194976位点多态分析及其与尾（臀）脂性状相关性研究[J]，基因组学与应用生物学, 32(5): 575-580.( Wang SY, Zhang W, Shen M, et al.2013. Polymorphism analysis of the position(59194976) on sheep X chromosome and its relationship with fat-tail(fat-rump) trait[J].Genomics and Applied Biology, 2013, 32(5): 575-580.)
[4]张伟, 沈敏, 李欢, 等. 2013. 绵羊X染色体59571364与59912586位点在脂尾、瘦尾绵羊群体中的多态性检测及分析[J]. 遗传, 35(12): 1384-1390. (Zhang W, Shen M, Li H, et al. 2013.Detection and analysis of polymorphisms of 59571364 and 59912586 loci on X chromosome in fat-tail and thin-tail sheep flocks[J]. Hereditas(Beiijng), 35(12): 1384-1390.)
[5]Auwerx J, Cock TA, and Knouff C. 2003. PPAR-gamma: a thrifty transcription factor[J]. Nuclear receptor signaling, (1):e006.
[6]Bing C, Gomez-Ambrosi J, Zabalegui N, et al. 2002. Resistin and RELM-alpha gene expression in white adipose tissue of lactating mice[J]. Biochemical and biophysical research communications, 296(2): 458-462.
[7]Gong H, Ni Y, Guo X, et al. 2004. Resistin promotes 3T3-L1 preadipocyte differentiation[J]. European Journal of Endocrinology, 150:885-892.
[8]Hartman HB, Hu X, Tyler KX, et al. 2002. Mechanisms regulating adipocyte expression of resistin[J]. The Journal of biological chemistry, 277(22): 19754-19761.
[9]Hivert MF, Sullivan LM, Fox CS, et al. 2008. Associations of adiponectin, resistin, and tumor necrosis factor-alpha with insulin resistance[J]. The Journal of clinical endocrinology and metabolism, 93(8): 3165-3172.
[10]Issemann I, and Green S. 1990. Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators[J]. Nature, 347(6294): 645-650.
[11]Kim KH, Lee K and Moon YS.2001. A cysteine-rich adipose tissues pecific secretory factor inhibits adipocyte differentiation [J[. The journal of Biological Chemistry, 276(14):11252-11256.
[12]Kitagawa Y, Bujo H, Takahashi K, et al. 2004. Impaired glucose tolerance is accompanied by decreased insulin sensitivity in tissues of mice implanted with cells that overexpress resistin [J]. Diabetologia, 47(10):1847-1853.
[13]Koebnick C, Wagner K, Garcia A L, et al. 2006. Increase in serum resistin during weight loss in overweight subjects is related to lipid metabolism [J].International Journal of Obesity, 30(7):1097-1103.
[14]Levy JR, Davenport B, Clore JN et al. 2002.Lipid metabolism and resistin gene expression in insulin resistant Fischer 344 rats[J]. American Journal of Physiology Endocrinology and Metabolism, 282(3):626-633.
[15]Li J, Yu X, Pan W, et al. 2002. Gene expression profile of rat adipose tissue at the onset of high-fat-diet obesity[J]. American journal of physiology Endocrinology and metabolism, 282(6): E1334-1341.
[16]Li ZH, Carraro R, Gregerman RI, et al. 1998. Adipocyte differentiation factor (ADF): a protein secreted by mature fat cells that induces preadipocyte differentiation in culture[J]. Cell biology international, 22(4): 253-270.
[17]Linhart HG, Ishimura-Oka K, DeMayo F, et al. 2001. C/EBPalpha is required for differentiation of white, but not brown, adipose tissue[J]. Proceedings of the National Academy of Sciences of the United States of America, 98(22): 12532-12537.
[18]Liu F, Guo XR, Gong HX, et al. 2006. A resistin binding peptide selected by phage display inhibits 3T3-L1 preadipocyte differentiation[J]. Chinese medical journal, 119(6): 496-503.
[19]McTernan CL, McTernan PG, Harte AL, et al. 2002. Resistin, central obesity, and type 2 diabetes[J]. Lancet, 359(9300): 46-47.
[20]Morash BA, Willkinson D, Ur E, et al. 2002. Resistin expression and regulation in mouse pituitary[J]. FEBS letters, 526(1-3): 26-30.
[21]Niederwanger A, Kranebitter M, Ciardi C, et al. 2007. Resistin impairs basal and insulin induced glycogen synthesis by different mechanisms[J]. Molecular and Cellular Endocrinology, 263 (12):112-119.
[22]Otto TC, and Lane MD. 2005. Adipose development: from stem cell to adipocyte[J]. Critical reviews in biochemistry and molecular biology, 40(4): 229-242.
[23]Sassek M, Pruszynska-Oszmalek E, Nowacka-Woszuk J, et al. 2013. Resistin - from gene expression to development of diabetes[J]. Journal of biological regulators and homeostatic agents, 27(3): 647-654.
[24]Seale P, Kajimura S, Yang W, et al. 2007. Transcriptional control of brown fat determination by PRDM16[J]. Cell metabolism, 6(1): 38-54.
[25]Shuldiner AR, Yang R, Gong DW. 2001.Resistin, obesity and insulin resistance-the emerging role of the adi- pocyte as an endocrine organ [J]. N Engl J Med, 345(18): 1345- 1346.
[26]Sliwicka E, Pilaczynska-Szczesniak L, Nowak A, et al. 2012. Resistin, visfatin and insulin sensitivity in selected phases of annual training cycle of triathletes[J]. Acta physiologica Hungarica, 99(1): 51-60.
[27]Steppan CM, and Lazar MA. 2004. The current biology of resistin[J]. Journal of internal medicine, 255(4): 439-447.
[28]Steppan CM, Bailey ST, Bhat S, et a1.2001. The hormone resistinlinks obesity to diabetes[J]. Nature, 409(6818): 307- 312.
[29]Steppan CM, Brown EJ, Wright CM, et al. 2001.A family of tissue-specific resistin-like molecules [J]. Proc Nat1 Acad Sci USA, 98(2): 502-506.
[30]Tontonoz P, Kim JB, Graves RA, et al. 1993. ADD1: a novel helix-loop-helix transcription factor associated with adipocyte determination and differentiation[J]. Molecular and cellular biology, 13(8): 4753-4759.
[31]Wu Z, Rosen ED, Brun R, et al. 1999. Cross-regulation of C/EBP alpha and PPAR gamma controls the transcriptional pathway of adipogenesis and insulin sensitivity[J]. Molecular Cell, 3(2): 151-158.
[32]Wu Z, Xie Y, Bucher NL, et al. 1995. Conditional ectopic expression of C/EBP beta in NIH-3T3 cells induces PPAR gamma and stimulates adipogenesis[J]. Genes Development, 9(19): 2350-2363.
[33]Yin Y, Yuan H, Wang C, et al. 2006. 3-phosphoinositide-dependent protein kinase-1 activates the peroxisome proliferator-activated receptor-gamma and promotes adipocyte differentiation[J]. Molecular endocrinology, 20(2): 268-278.
