2种多不饱和脂肪酸对樱桃谷鸭脂质和<i>SREBP</i>1基因表达的效应

罗华伦; 李万贵; 张依裕; 覃媛钰; 吴磊; 余蓉蓉

doi:10.19303/j.issn.1008-0384.2018.05.002

2种多不饱和脂肪酸对樱桃谷鸭脂质和SREBP1基因表达的效应

doi: 10.19303/j.issn.1008-0384.2018.05.002

罗华伦^{1, 2,},
李万贵³,
张依裕^{1, 2, ,},
覃媛钰^{1, 2},
吴磊^{1, 2},
余蓉蓉⁴

1.
贵州大学动物科学学院, 贵州贵阳 550025
2.
高原山地动物遗传育种与繁殖教育部重点实验室, 贵州贵阳 550025
3.
六盘水市草地工作站, 贵州六盘水 553001
4.
贵州省贵阳市花溪区农业局, 贵州贵阳 550025

基金项目:

国家自然科学基金项目 31160453

国家自然科学基金项目 31760663

详细信息

作者简介:
罗华伦(1993-), 女, 硕士研究生, 主要从事动物遗传资源评价、保护与利用研究(E-mail:463809726@qq.com)

通讯作者:
张依裕(1976-), 男, 副教授, 主要从事动物遗传资源评价、保护与利用研究(E-mail:zyy8yyc@163.com)

中图分类号: S834
计量
- 文章访问数: 1564
- HTML全文浏览量: 219
- PDF下载量: 9
- 被引次数: 0
出版历程
- 收稿日期: 2018-01-12
- 修回日期: 2018-04-02
- 刊出日期: 2018-05-01

Effects of Two Polyunsaturated Fatty Acids on Lipid Contents and SREBP1 Expression in Meat-type Ducks

LUO Hua-lun^{1, 2
,},
LI Wan-gui³,
ZHANG Yi-yu^{1, 2
, ,},
QIN Yuan-yu^{1, 2},
WU Lei^{1, 2},
YU Rong-rong⁴

1.
College of Animal Science, Guizhou University, Guiyang, Guizhou 550025, China
2.
Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education, Guiyang, Guizhou 550025, China
3.
The Grass Workstation of Liupanshui City, Liupanshui, Guizhou 553001, China
4.
Huaxi District Agricultrual Bureau, Guiyang, Guizhou 550025, China

摘要

摘要: 多不饱和脂肪酸（polyunsaturated fatty acids，PUFAs）主要通过调控脂肪生成基因和脂肪分解基因的表达来实现畜禽机体脂质的动态平衡。为了探明亚油酸和二十碳五烯酸（eicosapentaenoic acid，EPA）对肉鸭脂质和固醇调节元件结合1（sterol regulatory elemement binding protein 1，SREBP1）基因表达的影响，设置基础饲粮组、基础饲粮+4%亚油酸组、基础饲粮+4% EPA 3个处理饲喂樱桃谷母鸭，4、6、8和10周龄检测SREBP1 mRNA表达丰度和脂质含量，分析其表达与脂质指标的相关性。结果表明：饲粮中添加亚油酸和EPA可引起胸肌中饱和脂肪酸（saturated fatty acids，SFA）、多不饱和脂肪酸（PUFA）、必需脂肪酸（essential fatty acids，EFA）和血清甘油三酯（triglyceride，TG）、总胆固醇（total cholesterol，TCHO）、高密度脂蛋白（high density lipoprotein，HDL）的明显升高，说明2种脂肪酸对脂肪酸组成和脂质有影响，且亚油酸的影响效果强于二十碳五烯酸；引起SREBP1基因mRNA表达量下降，表明2种脂肪酸对SREBP1基因的转录具有抑制作用；SREBP1基因表达与腹脂、肌内脂肪（intramuscular fat，IMF）、不饱和脂肪酸（unsaturated acids，UFA）、多不饱和脂肪酸（PUFA）、必需脂肪酸（EFA）、甘油三酯（TG）、总胆固醇（TCHO）呈显著正相关（P < 0.05），表明SREBP1表达对脂质沉积有正向效应。综合结果揭示亚油酸和二十碳五烯酸与SREBP1基因互作调控肉鸭脂肪沉积。
- 肉鸭 /
- 亚油酸 /
- 二十碳五烯酸 /
- 脂质 /
- SREBP1基因
Abstract: Polyunsaturated fatty acids(PUFAs) play a critical role in animal lipid homeostasis by regulating the expressions of lipogenic and lipolytic genes. This study attempted to decipher the effects of linoleic acid and eicosapentaenoic acid (EPA) in diet on the lipids and the sterol regulatory element binding protein 1 (SREBP1) gene in meat-type ducks. Indoor-raised female Cherry Valley ducks were randomly divided into three groups to be fed each with either a basal diet, the basal diet+4% linoleic acid, or the basal diet+4% EPA. Content of lipids and abundance of SREBP1 mRNA were determined in 4, 6, 8 and 10 weeks after the feeding started. The addition of linoleic acid and EPA in the diet was found to raise the contents of saturated fatty acids, PUFAs and essential fatty acids (EFA) in the duck muscles as well as those of triglyceride (TG), total cholesterol (TCHO) and high-density lipoprotein (HDL) in the blood. The effect induced by the addition of linoleic acid was greater than that of EPA. The mRNA in liver SREBP1 declined with the added fatty acids suggesting their inhibitory effect on the transcription of the gene, with a slightly greater extent by EPA than linoleic acid. The expression of SREBP1 significantly correlated with the abdominal fat, intramuscular fat, unsaturated fatty acids, PUFA, EFA, serum TG and TCHO in the ducks. It appeared that linoleic acid, EPA and SREBP1 gene regulation interacted to affect the lipid deposit in the ducks.
- meat-type duck /
- linoleic acid /
- eicosapentaenoic acid /
- lipids /
- SREBP1 gene

HTML全文

图 1 总RNA提取与引物扩增效果

注：A为总RNA；B为β-Actin；C为SREBP1。

Figure 1. Detection of total RNA and amplified primer

下载: 全尺寸图片幻灯片

图 2 β-Actin(左)和SREBP1(右)基因实时荧光定量PCR扩增熔解曲线

Figure 2. qRT-PCR melting cures of β-actin (left) and SREBP1 gene (right)

下载: 全尺寸图片幻灯片

表 1 基础饲粮原料组成及营养水平

Table 1. Dietary composition and nutrients in feed

原料	含量/%	营养水平	含量
玉米	60.00	代谢能/(MJ·kg^-1)	11.53
豆粕	19.40	粗蛋白质/%	18.47
麸皮	11.70	钙/%	1.10
棉籽饼	3.00	总磷/%	0.77
鱼粉	3.00	蛋氨酸/%	0.27
磷酸氢钙	1.50	赖氨酸/%	0.97
石粉	1.10
食盐	0.30
合计	100.00

下载: 导出CSV

表 2 不同处理樱桃谷鸭脂肪沉积比较

Table 2. Deposit of indicator lipids on treated ducks

项目	Ⅰ组	Ⅱ组	Ⅲ组
腹脂/%	2.26±0.44	2.52±0.33	2.67±0.42
肌内脂肪/%	8.56±1.15	9.01±0.46	8.87±0.44
饱和脂肪酸/%	43.98±0.33a	42.14±0.20b	42.92±0.37ab
不饱和脂肪酸/%	57.03±0.47	57.87±0.50	57.09±0.43
多不饱和脂肪酸/%	19.65±0.23a	22.87±0.20b	21.01±0.25ab
必需脂肪酸/%	19.37±0.32a	22.29±0.19b	20.57±0.24ab
甘油三酯/(mmol·L^-1)	0.79±0.13a	1.10±0.19b	1.02±0.14b
总胆固醇/(mmol·L^-1)	4.46±0.22a	5.68±0.17b	5.26±0.13ab
高密度脂蛋白/(mmol·L^-1)	2.77±0.21a	3.98±0.32b	3.67±0.27ab
低密度脂蛋白/(mmol·L^-1)	1.78±0.15	1.91±0.24	1.83±0.11
注：同行数据后不同小写字母表示差异显著(P < 0.05)。表 3同。

下载: 导出CSV

表 3 SREBP1基因在樱桃谷鸭肝脏中不同时期mRNA表达水平

Table 3. mRNA level of liver SREBP1 gene in Cherry Valley ducks at various growth periods

周龄	Ⅰ组	Ⅱ组	Ⅲ组
4	1.77±0.11a	1.21±0.18b	1.17±0.13b
6	2.23±0.13a	1.79±0.19b	1.73±0.14b
8	2.45±0.21a	2.13±0.15b	1.98±0.17b
10	4.88±0.14a	2.46±0.12b	2.34±0.13b

下载: 导出CSV

表 4 SREBP1基因表达与樱桃谷鸭脂质指标的相关性

Table 4. Correlation between SREBP1 gene expression and indicator lipids of Cherry Valley ducks

指标	相关系数
腹脂/%	0.78^*
肌内脂肪/%	0.67^*
饱和脂肪酸/%	0.13
不饱和脂肪酸/%	0.47^*
多不饱和脂肪酸/%	0.51^*
必需脂肪酸/%	0.48^*
甘油三酯/(mmol·L^-1)	0.67^*
总胆固醇/(mmol·L^-1)	0.53^*
高密度脂蛋白/(mmol·L^-1)	0.29
低密度脂蛋白/(mmo·L^-1)	0.33
注：“*”表示相关性达显著水平(P < 0.05)。

下载: 导出CSV

参考文献(27)

[1]	HAN J, LI E, CHEN L, et al. The CREB coactivator CRTC2 controls hepatic lipid metabolism by regulating SREBP1[J]. Nature, 2015, 524(7564):243-246. doi: 10.1038/nature14557
[2]	YEN C F, JIANG Y N, SHEN T F, et al. Cloning and expression of the genes associated with lipid metabolism in Tsaiya ducks[J]. Poultry Science, 2005, 84(1):67-74. doi: 10.1093/ps/84.1.67
[3]	FLEISCHMANN M, IYNEDJIAN P B. Regulation of sterol regulatory-element binding protein 1 gene expression in liver:role of insulin and protein kinase B/cAkt[J]. Biochemical Journal, 2000, 349(1):13-17. doi: 10.1042/bj3490013
[4]	WANG Y, VISCARRA J, KIM S J, et al. Transcriptional regulation of hepatic lipogenesis[J]. Nature reviews Molecular cell biology, 2015, 16(11):678-689. doi: 10.1038/nrm4074
[5]	CLARKE S D. Polyunsaturated fatty acid regulation of gene transcription:a molecular mechanism to improve the metabolic syndrome.[J]. Journal of Nutrition, 2001, 131(4):1129-1132. doi: 10.1093/jn/131.4.1129
[6]	KOBATAKE K, SAKO K, IZAWA M, et al. Autora-diographic determination of brain pH following middle cerebral artery occlusion in the rat[J]. Stroke:A Journal of Cerebral Circulation, 1984, 15(3):540. doi: 10.1161/01.STR.15.3.540
[7]	KERSTEN S, SSYDOUX J, PETERS J M, et al. Peroxisome proliferator-activated receptor alpha mediates the adaptive response to fasting[J]. Journal of Clinical Investigation, 1999, 103(11):1489-1498. doi: 10.1172/JCI6223
[8]	SAMPATH H, NTAMBI J M. Polyunsaturated fatty acid regulation of gene expression[J]. Nutrition reviews, 2004, 62(9):333-339. doi: 10.1111/nure.2004.62.issue-9
[9]	MAEKAWA M, WATANABE A, IWAYAMA Y, et al. Polyunsaturated fatty acid deficiency during neurodevelopment in mice models the prodromal state of schizophrenia through epigenetic changes in nuclear receptor genes[J]. Transl Psychiatry, 2017, 7(9):1229. doi: 10.1038/tp.2017.182
[10]	JIN M, MONROIG Í, LU Y, et al. Dietary DHA/EPA ratio affected tissue fatty acid profiles, antioxidant capacity, hematological characteristics and expression of lipid-related genes but not growth in juvenile black seabream (Acanthopagrus schlegelii)[J]. PLoS One, 2017, 12(4):e0176216. doi: 10.1371/journal.pone.0176216
[11]	兰云贤.动物饲养标准[M].昆明:云南师范大学出版社, 2008:80-81.
[12]	中华人民共和国农业部. 家禽生产性能名词术语和度量统计方法: NY/T823-2004[S]. 北京: 中国标准出版社, 2004.
[13]	LIAO T, WANG Y J, HU J Q, et al. Histone methyltransferase KMT5A gene modulates oncogenesis and lipid metabolism of papillary thyroid cancer in vitro[J]. Oncology Reports, 2018, 39(5):2185-2192. http://cn.bing.com/academic/profile?id=f44bf194fc56c79dc56fb61084a229d4&encoded=0&v=paper_preview&mkt=zh-cn
[14]	WANG L F, WANG X N, HUANG C C, et al. Inhibition of NAMPT aggravates high fat diet-induced hepatic steatosis in mice through regulating Sirt1/AMPKα/SREBP1 signaling pathway[J]. Lipids in health and disease, 2017, 16(1):82-95. doi: 10.1186/s12944-017-0464-z
[15]	YIN F, SHAREN G, YUAN F, et al. TIP30 regulates lipid metabolism in hepatocellular carcinoma by regulating SREBP1 through the Akt/mTOR signaling pathway[J]. Oncogenesis, 2017, 6(6):347. doi: 10.1038/oncsis.2017.49
[16]	SIQINGAOWA, SEKAR S, GOPALAKRISHNAN V, et al. Sterol regulatory element-binding protein 1 inhibitors decrease pancreatic cancer cell viability and proliferation[J]. Biochemical and biophysical research communications, 2017, 488(1):136-140. doi: 10.1016/j.bbrc.2017.05.023
[17]	FU R Q, LIU R R, ZHAO G P, et al. Expression profiles of key transcription factors involved in lipid metabolism in Beijing-You chickens[J]. Gene, 2014, 537(1):120-125. doi: 10.1016/j.gene.2013.07.109
[18]	YANG Y, YANG Q, YANG J, et al. Angiotensin Ⅱ induces cholesterol accumulation and injury in podocytes[J]. Scientific Reports, 2017, 7(1):10672-10682. doi: 10.1038/s41598-017-09733-w
[19]	RUAN D, LIN Y C, CHEN W, et al. Effects of rice bran on performance, egg quality, oxidative status, yolk fatty acid composition, and fatty acid metabolism-related gene expression in laying ducks[J]. Poultry Science, 2015, 94(12):2944-2951. doi: 10.3382/ps/pev286
[20]	ZHANG L, LI C, WANG F, et al. Treatment with PPARα Agonist Clofibrate Inhibits the Transcription and Activation of SREBPs and Reduces Triglyceride and Cholesterol Levels in Liver of Broiler Chickens[J]. PPAR Research, 2015(5):1-10. http://cn.bing.com/academic/profile?id=d12b3fc1a7dca4a4dd14b09b8fc82bd8&encoded=0&v=paper_preview&mkt=zh-cn
[21]	LI Q, ZHAO X L, GILBERT E R, et al. Confined housing system increased abdominal and subcutaneous fat deposition and gene expressions of carbohydrate response element-binding protein and sterol regulatory element-binding protein 1 in chicken[J]. Genetics and Molecular Research, 2015, 14(1):1220-1228. doi: 10.4238/2015.February.6.24
[22]	WANG G, WILLIAMS C A, McCONN B R, et al. A high fat diet enhances the sensitivity of chick adipose tissue to the effects of centrally injected neuropeptide Y on gene expression of adipogenesis-associated factors[J]. Comparative Biochemistry and Physiology Part A Molecular & Integrative Physiology, 2017, 211(9):49-55. http://cn.bing.com/academic/profile?id=ae139a0a7511d10c7202bf11ed5a0b5d&encoded=0&v=paper_preview&mkt=zh-cn
[23]	皮宇, 经语佳, 王梦芝, 等.不同脂肪酸对体外培养的瘤胃微生物活力和蛋白质含量的影响[J].动物营养学报, 2014, 26(1):260-269. http://www.cqvip.com/QK/98510X/201401/48510104.html
[24]	JUMP D B, BOTOLIN D, WANG Y, et al. Docosa-hexaenoic acid (DHA) and hepatic gene transcription[J]. Chemistry and Physics of Lipids, 2008, 153(1):3-13. doi: 10.1016/j.chemphyslip.2008.02.007
[25]	YOSHIKAWA T, SHIMANO H, YAHAGI N, et al. Polyunsaturated fatty acids suppress sterol regulatory element-binding protein 1c promoter activity by inhibition of liver X receptor (LXR) binding to LXR response elements[J]. The Journal of Biological Chemistry, 2002, 277(3):1705-1711. doi: 10.1074/jbc.M105711200
[26]	NAKAMURA M T, CHEON Y, LI Y, et al. Mechanisms of regulation of gene expression by fatty acids[J]. Lipids, 2004, 39(11):1077-1083. doi: 10.1007/s11745-004-1333-0
[27]	MUHLHAUSLER B S, COOK-JOHNSON R, JAMES M, et al. Opposing effects of omega-3 and omega-6 long chain polyunsaturated Fatty acids on the expression of lipogenic genes in omental and retroperitoneal adipose depots in the rat[J]. Journal of Nutrition and Metabolism, 2010(4):927836-927845. http://cn.bing.com/academic/profile?id=e8e274b589d58b195996df6834bb77df&encoded=0&v=paper_preview&mkt=zh-cn