绵羊FAM134B、PPARγ、HSL和FAS基因表达量及与肌内脂肪含量的关系

doi:10.11843/j.issn.0366-6964.2016.12.007

摘要/Abstract

摘要：

旨在探究FAM134B、PPARγ、FAS和HSL基因在绵羊肌肉发育过程中的组织表达规律及其与肌内脂肪（Intramuscular fat, IMF）含量的关系，以期为研究绵羊IMF沉积的分子调控机制奠定理论基础。试验选取2、4、5、6和12月龄敖汉细毛羊公羊各5只，屠宰，采集背最长肌和股二头肌测定其IMF含量，应用实时荧光定量 PCR技术检测PPARγ、FAM134B、FAS和HSL基因在肌肉不同发育时期的mRNA表达量，并对基因表达量之间及表达量与IMF含量的关系进行分析。结果表明：（1）2~5月龄时，背最长肌和股二头肌的IMF含量均随着月龄的增加而增加；而5~12月龄时则基本保持不变；同月龄背最长肌IMF含量极显著高于股二头肌（P<0.01）。（2）背最长肌中FAM134B表达量呈上升-下降趋势，6月龄最高（P<0.01）；股二头肌中FAM134B表达量呈先上升后稳定趋势。FAS基因在两部位肌肉中表达量均呈上升趋势。PPARγ基因在两部位肌肉中表达量均呈下降-上升趋势。HSL基因在两部位肌肉中表达量均呈下降-上升-下降趋势。同月龄目的基因表达量在两部位肌肉中均表现出明显的差异。（3）关联分析显示，两部位肌肉中FAM134B与FAS表达量均呈显著正相关（P<0.05），与PPARγ、HSL表达量呈不同程度负相关。背最长肌中，FAM134B、FAS表达量与IMF含量呈极显著或显著正相关（P<0.01，P<0.05），PPARγ表达量与IMF含量显著负相关（P<0.05）；股二头肌中，FAM134B表达量与IMF含量显著正相关（P<0.05）。综上，FAM134B、FAS可能会促进IMF的沉积，而PPARγ、HSL对IMF的沉积可能具有抑制作用；FAM134B可能通过刺激脂肪合成基因的表达，而抑制脂肪分解基因的表达来调控IMF的含量。结果可为进一步解析相关基因的功能和调控IMF沉积的分子机制提供参考。

关键词: 肌内脂肪, 基因, 表达, 相关性

Abstract:

The objectives of this study were to investigate the developmental changes of FAM134B, PPARγ, FAS and HSL gene expression in the process of sheep muscles development and their association with intramuscular fat (IMF) accumulation, to lay a theoretical foundation for studying molecular regulation mechanism of the IMF deposition in sheep. Five Aohan fine wool sheep were slaughtered at 2, 4, 5, 6 and 12 months old to collect samples from longissimus dorsi and biceps femoris, for the purpose of determining the IMF content. Then mRNA expression level of the FAM134B, PPARγ, FAS and HSL gene in different development period of muscles were analyzed by real-time PCR, and association between genes expression and IMF contents were analyzed. The results showed that: (1) Among 2-5 months old, with growing of the age, the IMF contents of longissimus dorsi and biceps femoris were increasing. Among 5-12 months old, the IMF contents remained unchanged; The IMF content of longissimus dorsi were extremely significantly higher than biceps femoris (P<0.01) at the same age. (2) In longissimus dorsi, FAM134B expression level presented first increased and then declined with growing, the highest was in 6 months old(P<0.01). In biceps femoris,FAM134B expression level presented increased and then stabilized with growing. FAS expression level increased with growing in both of two parts of muscles. PPARγ expression level presented first declined and then increased with growing. HSL expression levels was showed “declined-increased-declined” patterns in two parts of muscles. In the same month, target genes expression level had tissue-dependent in two parts of muscles.(3) Correlation analysis showed that the expression level of FAM134B was significantly positively related to FAS in two parts of muscles(P<0.05), and was negatively related to PPARγ and HSL expression levels at different levels. In longissimus dorsi, FAM134B, FAS expression levels were significantly positively related to IMF content(P<0.01 or P<0.05, respectively), PPARγ expression level was significantly negatively related to IMF content(P<0.05). In biceps femoris, FAM134B expression level was significantly positively correlated with the IMF content(P<0.05). In conclusion, FAM134B, FAS expression might have positively effect on IMF accumulation, and PPARγ, HSL expression might have negatively effect on IMF accumulation. FAM134B was likely to regulate IMF content through stimulating expression level of fat synthetic genes and inhibiting adipose decompose genes. The results would provide reference for studying related gene functions and molecular mechanism of the IMF deposition.

Key words: IMF, gene, expression, correlation

中图分类号:

S826
S813.2

栾兆进，刘开东，贺建宁，程明，曲绪仙，柳楠. 绵羊FAM134B、PPARγ、HSL和FAS基因表达量及与肌内脂肪含量的关系[J]. 畜牧兽医学报, 2016, 47(12): 2379-2389.

LUAN Zhao-jin, LIU Kai-dong, HE Jian-ning, CHENG Ming, QU Xu-xian, LIU Nan. The FAM134B, PPARγ, HSL and FAS Gene Expression Patterns and Their Association with Intramuscular Fat Content in Sheep[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(12): 2379-2389.

参考文献

［1］URBAN T, MIKOLÁSOVÁ R, KUCIEL J, et al. A study of associations of the H-FABP genotypes with fat and meat production of pigs［J］. J Appl Genet, 2002, 43(4): 505-509.
［2］杜琛，李金泉，陈秀娟.白绒山羊 PPARγ基因RNA干扰慢病毒载体的构建及对肌内脂肪细胞增殖分化的影响［J］.畜牧兽医学报，2016，47(4):671-678.
DU C, LI J Q, CHEN X J. Construction of lentiviral RNAi vector of PPARγ gene in cashmere goat and its effect on proliferation and differentiation of intramuscular adipocytes［J］. Acta Veterinaria et Zootechnica Sinica, 2016, 47(4):671-678.(in Chinese)
［3］WANG Y H, BYRNE K A, REVERTER A, et al. Transcriptional profiling of skeletal muscle tissue from two breeds of cattle［J］. Mamm Genome, 2005, 16(3): 201-210.
［4］赵有璋.羊生产学［M］.北京：中国农业出版社，2002:69-75.
ZHAO Y Z. Production science of sheep and goat［M］.Beijing: China Agriculture Press, 2002:69-75.(in Chinese)
［5］郝称莉，李齐发，乔永，等.湖羊肌肉组织H-FABP和PPARγ基因表达水平与肌内脂肪含量的相关研究［J］.中国农业科学，2008，41(11):3776-3783.
HAO C L, LI Q F, QIAO Y, et al. Association of the H-FABP and PPARγ gene expression with intramuscular fat content in Hu sheep muscles［J］. Scientia Agricultura Sinica, 2008, 41(11):3776-3783.(in Chinese)
［6］MEUWISSEN T H E, GODDARD M E. The use of marker haplotypes in animal breeding schemes［J］. Genet Sel Evol, 1996, 28:161-176.
［7］KURTH I, PAMMINGER T, HENNINGS J C, et al. Mutations in FAM134B,encoding a newly identified Golgi protein,cause severe sensory and autonomic neuropathy［J］. Nat Genet, 2009, 41(11): 1179-1181.
［8］朱武政，江明锋，唐璐, 等.山羊FAM134B基因克隆、序列分析及其mRNA表达与肌内脂肪含量的相关性［J］.农业生物技术学报，2014，22(10):1277-1285.
ZHU W Z, JIANG M F, TANG L, et al. Cloning,sequence analysis of FAM134B and correlation analysis between its mRNA expression and intramuscular fat content in goat (Capra hirus)［J］.Journal of Agricultural Biotechnology, 2014, 22(10):1277-1285.(in Chinese)
［9］GRAUGNARD D E, PIANTONI P, BIONAZ M, et al. Adipogenic and energy metabolism gene networks in longissimus lumborum during rapid post- weaning growth in Angus and Angus×Simmental cattle fed high- starch or low-starch diets［J］. BMC Genomics, 2009, 10:142.
［10］TONTONOZ P, NAGY L, ALVAREZ J G, et al. PPARγ promotes monocyte/macrophage differentiation and uptake of oxidized LDL［J］. Cell, 1998, 93(2): 241-252.
［11］QIAO Y, HUANG Z, LI Q, et al. Developmental changes of the FAS and HSL mRNA expression and their effects on the content of intramuscular fat in kazak and xinjiang sheep［J］. J Genet Genomics, 2007, 34(10): 909-917.
［12］颜新春, 汪以真, 许梓荣. 日粮因素对脂肪酸合成酶 (FAS) 基因表达调控的影响及其应用［J］.上海畜牧兽医通讯，2000，4:7-9.
YAN X C, WANG Y Z, XU Z R. Regulation of animal fatty acid synthase (FAS) gene expression［J］. Shanghai Journal of Animal Husbandry and Veterinary Medicine, 2000, 4:7-9.(in Chinese)
［13］XU H, XU G, WANG D, et al. Molecular cloning, sequence identification and expression analysis of novel caprine MYLPF gene［J］. Mol Biol Rep, 2013, 40(3):2565-2572.
［14］DA COSTA A S, PIRES V M, FONTES C M, et al. Expression of genes controlling fat deposition in two genetically diverse beef cattle breeds fed high or low silage diets［J］. BMC Vet Res, 2013, 9: 118.
［15］ILGAZ AYDINLAR E, ROLFS A, SERTESER M, et al. Mutation in FAM134B causing hereditary sensory neuropathy with spasticity in a Turkish family［J］. Muscle Nerve, 2014, 49(5):774-775.
［16］MURPHY S M, DAVIDSON G L, BRANDNER S, et al. Mutation in FAM134B causing severe hereditary sensory neuropathy［J］. J Neurol Neurosurg Psychiatry, 2012, 83(1):119-120 .
［17］袁章琴，汪以真.猪新基因FAM134B基因对肌内脂肪沉积及其分子机理研究［J］.畜牧与兽医，2012，44(S1):162-163.
YUAN Z Q, WANG Y Z. Association of FAM134B in pig for intramuscular fat deposition and its molecular mechanism［J］. Animal Husbandry And Veterinary Medicine, 2012, 44(S1):162-163.(in Chinese)
［18］NADEAU K J, EHLERS L B, AGUIRRE L E, et al. Exercise training and calorie restriction increase SREBP-1 expression and intramuscular triglyceride in skeletal muscle［J］. Am J Physiol Endocrinol Metab, 2006, 291(l):E90-98.
［19］刘桂，殷亮，王晓慧，等.高脂饮食诱导的肥胖与肥胖抵抗大鼠FAS和ACAT-2的蛋白表达差异［J］.上海体育学院学报，2014，38(6):105-109.
LIU G, YIN L, WANG X H, et al. Differential expression of FAS and ACAT-2 in livers of high fat diet induced obesity and obesity resistant rat［J］.Journal of Shanghai University of Sport, 2014, 38(6):105-109.(in Chinese)
［20］舒常平，王宝维，李桢, 等. 填饲鹅肝脏组织中脂肪酸沉积与FAS基因mRNA的表达丰度［J］.中国农业科学，2012，45(10):2002-2011.
SHU C P, WANG B W, LI Z, et al. Fatty acids deposition and FAS mRNA expression abundance in liver tissue of overfeeding goose［J］. Scientia Agricultura Sinica, 2012,45(10):2002-2011. (in Chinese)
［21］SORET B, LEE H J, FINLEY E, et al. Regulation of differentiation of sheep subcutaneous and abdominal preadipocytes in culture［J］. J Endocrinol, 1999, 161(3): 517-524.
［22］REBECCA E, STEVEN M, KEITH I, et al. An investigation into the genetic controls of pork quality［R］. National Swine Improvement Federation, 2000.
［23］姚艳，王宗保，唐朝克. PPARs在长链脂肪酸调控能量代谢中的作用［J］.生理科学进展，2016，47(1):1-6.
YAO Y, WANG Z B, TANG C K. PPARs mediate the regulation of energy metabolism by long-chain fatty acids［J］. Progress in Physiological Sciences, 2016, 47(1):1-6.(in Chinese)
［24］郝称莉.湖羊肌肉肌内脂肪相关基因表达的发育性变化及与肌内脂肪含量的相关性研究［D］.南京: 南京农业大学，2008.
HAO C L. Study on the developmental changes of the genes related to intramuscular fat expression level in muscles in Hu sheep and association with intramuscular fat content［D］. Nanjing: Nanjing Agricultural University, 2008.(in Chinese)
［25］孙文星，南文婷，谷淑华, 等. miR-27b及其靶基因PPARγ 在肌内和皮下脂肪细胞中的差异表达分析［J］.南京农业大学学报，2016，39(2):249-254.
SUN W X, NAN W T, GU S H, et al. Differential expression of miＲ- 27b and its target gene PPARγ in intramuscular and subcutaneous adipocytes［J］. Journal of Nanjing Agricultural University, 2016, 39(2):249-254.(in Chinese)
［26］王倩，尹逊河，王元虎, 等.长白山野猪与杜洛克猪杂交1代猪肌内脂肪代谢酶HSL和MDH活性的发育性变化规律及其对肌内脂肪沉积影响的研究［J］.中国畜牧兽医，2013，40(12):121-124.
WANG Q, YIN X H, WANG Y H, et al. The development changes of HSL and MDH activities in muscle and their association with intramuscular fat in mount Changbai boar×Duroc pigs［J］. China Animal Husbandry of Veterinary Medicine, 2013, 40(12):121-124. (in Chinese)

[1]	欧阳清渊, 王郁石, 胡深强, 李亮, 刘贺贺, 何桦, 王继文. GATA4/5/6基因克隆及其在出生后鸭心脏和肝脏中的发育性表达模式[J]. 畜牧兽医学报, 2019, 50(9): 1775-1786.
[2]	滑留帅, 辛晓玲, 王璟, 罗生金, 曾滔, 巴提玛, 施巧婷, 徐照学, 王二耀. 利用混池重测序鉴定萨福克绵羊超数排卵关键调控基因[J]. 畜牧兽医学报, 2019, 50(9): 1813-1821.
[3]	宋慧子, 栾兆进, 王兆琛, 杜炜, 赵勇超, 张家新. 永生化绵羊附睾上皮细胞系的建立及其生物学特性分析[J]. 畜牧兽医学报, 2019, 50(9): 1822-1831.
[4]	王正荣, 张艳艳, 马勋, 刘乙, 徐梦飞, 孟季蒙, 薄新文. 细粒棘球绦虫原头蚴lncRNA表达谱分析[J]. 畜牧兽医学报, 2019, 50(9): 1864-1873.
[5]	高俊莹, 张东超, 李璇, 王华琳, 姜宁. 弓形虫表面抗原SAG1黏附宿主细胞表面硫化肝素的特性研究[J]. 畜牧兽医学报, 2019, 50(9): 1874-1881.
[6]	王盼举, 樊新丽, 张梦圆, 宋晶晶, 李敏, 吾力江, 巴音查汗. 地方株马驽巴贝斯虫Bc48截短体单克隆抗体的制备及应用[J]. 畜牧兽医学报, 2019, 50(9): 1882-1887.
[7]	凌英会, 朱露, 吴昊, 陈青, 权青, 刘勇, 李文雍, 张运海. 山羊FST慢病毒载体构建及其对卵巢卵泡颗粒细胞增殖的影响[J]. 畜牧兽医学报, 2019, 50(9): 1888-1896.
[8]	赵凯, 崔燕, 刘鹏刚, 何俊峰, 宋亮丽, 廖博, 杨晓晴. IL-17和HSP90在不同牛肝中的定位和定量研究[J]. 畜牧兽医学报, 2019, 50(9): 1897-1903.
[9]	邓天宇, 樊红樱, 杜立新, 赵福平, 王立贤. 呼伦贝尔羊重要脂肪相关基因mRNA差异表达分析[J]. 畜牧兽医学报, 2019, 50(9): 1926-1935.
[10]	李华振, 狄冉, 刘秋月, 胡文萍, 王翔宇, 马琳, 刘武军, 储明星. 类视黄醇X受体基因调控动物季节性发情和发情周期的研究进展[J]. 畜牧兽医学报, 2019, 50(8): 1525-1535.
[11]	刘家鑫, 魏霞, 邓天宇, 谢锐, 韩建林, 杜立新, 赵福平, 王立贤. 绵羊全基因组ROH检测及候选基因鉴定[J]. 畜牧兽医学报, 2019, 50(8): 1554-1566.
[12]	温东旭, 张雷, 索朗斯珠, 牛家强, 王玉恒, 贡嘎, 旦巴次仁, 席广银, 郭敏, 徐业芬. 牦牛Nramp1基因mRNA组织表达谱及其相关miRNAs初步研究[J]. 畜牧兽医学报, 2019, 50(8): 1576-1586.
[13]	吴海波, 边雪娇, 贾丽玲, 索伦. 二代靶向测序在CRISPR/Cas9靶区筛选中的应用性研究[J]. 畜牧兽医学报, 2019, 50(8): 1587-1595.
[14]	彭孝坤, 张宇, 黄晓瑜, 周广琛, 邢晓南, 张恩平. 急性冷应激对绵羊免疫功能和不同组织热休克蛋白70家族基因表达的影响[J]. 畜牧兽医学报, 2019, 50(8): 1625-1634.
[15]	王素华, 帅江冰, 李舟, 袁淑辉, 吴绍强, 吕继洲, 赵治国. 炭疽杆菌双重荧光定量PCR检测技术的建立和应用[J]. 畜牧兽医学报, 2019, 50(8): 1658-1665.