静脉滴注心房利钠肽对阿勒泰羊血液脂代谢激素的影响及尾脂转录组分析

doi:10.11843/j.issn.0366-6964.2021.011.009

摘要/Abstract

摘要： 旨在研究静脉滴注心房利钠肽（ANP）对绵羊血液中脂代谢激素及尾脂转录组的影响，为阐明ANP在调节绵羊脂代谢中的作用机理提供参考。本试验选取体重为（45.6±6.5） kg、健康状况良好、1.5岁阿勒泰母羊8只，试验采用自身对照设计，对照期颈静脉滴注100 mL生理盐水45 min；试验期以1.125 μg·kg^-1 BW颈静脉滴注100 mL ANP生理盐水溶液45 min，连续静脉滴注ANP 4 d，各期均在试验结束当天（即第4天）滴注开始后0、15、30、45、60、90和120 min采集血液，分离血浆，测定ANP、脂联素、胰岛素、瘦素和环鸟苷酸水平；每期采集尾脂，进行转录组测序及生物学信息分析，并采用实时荧光定量PCR技术检测候选基因相对表达量的变化。结果显示：1）静脉滴注ANP后，较对照期，试验期血浆ANP含量在30、90 min时显著增加（P<0.05）；血浆脂联素含量在30 min时显著增加（P<0.05），在90 min时极显著增加（P<0.01）；cGMP含量在30、90 min时极显著增加（P<0.01），在45、60 min时显著增加（P<0.05）；胰岛素含量在0、30、120 min时极显著增加（P<0.01），在15、45、60、90 min时显著增加（P<0.05）；瘦素含量在0、15 min时显著增加（P<0.05），在30 min时极显著增加（P<0.01）。2）转录组分析表明，共3 686个差异表达基因，其中1 482个基因上调表达、2 204个基因下调表达；GO功能富集分析显示，差异基因显著富集到118个生物学功能，主要与线粒体呼吸链和氧化磷酸化有关；KEGG通路分析表明，差异基因显著富集到46条通路中，主要参与核糖体、产热、氧化磷酸化和调节脂肪细胞中的脂肪分解等信号通路；qRT-PCR分析结果表明，AQP7、FABP4、PLIN5、ADIPOR2、MGLL、IDE和ACSL1表达趋势与RNA-seq结果一致。由此可见，外源ANP通过改变脂代谢相关激素水平及增加绵羊脂肪组织氧化磷酸化和产热促进脂肪分解。

关键词: 心房利钠肽, 脂肪代谢, 转录组测序, 尾脂, 脂解

Abstract: The aim of this study was to investigate the effects of intravenous infusion of atrial natriuretic peptide (ANP) on blood lipid metabolism hormones and tail lipid transcriptome of sheep, and to provide reference for elucidating the mechanism of ANP in regulating lipid metabolism in sheep. Eight healthy Altay ewes with the age of 1.5-year-old, body weight (BW) of (45.6±6.5)kg were selected, and self control design was used. During the control period, 100 mL saline was infused intravenously in 45 min, and during the trial period, 100 mL of ANP saline solution with a dose of 1.125 μg·kg^-1BW was infused intravenously in 45 min. Intravenous infusion lasted for 4 days. At the end of each period (4th day), blood samples were collected at 0, 15, 30, 45, 60, 90 and 120 min after infusion, and the content of ANP, adiponectin, insulin, leptin and cyclic guanosine (cGMP) in plasma were deterimed. The sheep tail fat samples were collected for trans-criptome sequencing and biological information analysis, and qRT-PCR was used to detect the change of relative candidate genes expression. The results showed that: 1) After intravenous infusion of ANP, compared to the control period, the plasma ANP level at 30 and 90 min was significantly higher during the trial period(P<0.05); Adiponectin level at 30 min significantly increased (P<0.05), and at 90 min extremely significantly increased (P<0.01); The plasma cGMP level at 30 and 90 min extremely significantly increased(P<0.01), and at 45 and 60 min significantly increased (P<0.05); The plasma insulin at 0, 30 and 120 min extremely significantly increased(P<0.01), at 15, 45, 60 and 90 min significantly increased (P<0.05); The plasma leptin level at 0 and 15 min significantly increased(P<0.05), at 30 min extremely significantly increased(P<0.01). 2) Transcriptome analysis showed that there were 3 686 differentially expressed genes, of which 1 482 genes were up-regulated and 2 204 genes were down-regulated; GO function enrichment analysis showed that differentially expressed genes were significantly enriched into 118 biological functions, mainly related to mitochondrial respiratory chain and oxidative phosphorylation; KEGG pathways analysis revealed that differentially expressed genes were significantly enriched into 46 pathways, mainly involved in ribosome, thermogenesis, oxidative phosphorylation and regulating lipolysis in adipocytes and other signaling pathways; qRT-PCR analysis results showed that the expression trends of AQP7, FABP4, PLIN5, ADIPOR2, MGLL,IDE and ACSL1 were consistent with the RNA-seq results, which proved that the trans-criptome sequencing data was reliable. In conclusion, exogenous ANP promotes lipolysis by changing the level of lipid metabolism-related hormones and increasing the oxidative phosphorylation and thermogenesis of adipose tissue in sheep.

Key words: atrial natriuretic peptide, fat metabolism, RNA-seq technology, tail fat, lipolysis

中图分类号:

S826.2

马昕, 杨光, 李翔宇, 刘越越, 陈勇. 静脉滴注心房利钠肽对阿勒泰羊血液脂代谢激素的影响及尾脂转录组分析[J]. 畜牧兽医学报, 2021, 52(11): 3076-3088.

MA Xin, YANG Guang, LI Xiangyu, LIU Yueyue, CHEN Yong. Effects of Intravenous Infusion of Atrial Natriuretic Peptide on Blood Lipid Metabolism Hormones and Transcriptome Analysis of Tail Adipose Tissue of Altay Sheep[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(11): 3076-3088.

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