低温对合作猪脂肪组织形态、脂代谢相关基因表达和酶活性及AMPK/PGC-1α通路的影响

doi:10.11843/j.issn.0366-6964.2024.08.015

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (8): 3418-3426.doi: 10.11843/j.issn.0366-6964.2024.08.015

低温对合作猪脂肪组织形态、脂代谢相关基因表达和酶活性及AMPK/PGC-1α通路的影响

李瑶¹(), 贾蕊¹, 李杰¹, 滚双宝¹^,²^,³^,*(), 杨巧丽¹^,³^,*(), 王龙龙¹, 张鹏霞¹, 高小莉¹, 黄晓宇¹

1. 甘肃农业大学动物科学技术学院，兰州 730070
2. 甘肃省现代养猪工程技术研究中心，兰州 730070
3. 甘肃迭部蕨麻猪科技小院，甘南 740070

收稿日期:2024-01-30 出版日期:2024-08-23 发布日期:2024-08-28
通讯作者: 滚双宝,杨巧丽 E-mail:LY926111@qq.com;gunsb@gsau.edu.cn;yangql0112@163.com
作者简介:李瑶(2001-)，女，甘肃甘谷人，硕士生，主要从事动物遗传育种与繁殖研究，E-mail: LY926111@qq.com
基金资助:
国家自然科学基金区域创新发展联合基金项目(U22A20507);甘肃农业大学畜牧学养猪产业技术创新团队项目(GAU-XKTD-2022-25)

Effects of Low Temperature on Adipose Tissue Morphology, Lipid Metabolism-Related Gene Expression and Enzyme Activities, and AMPK/PGC-1α Pathway in Hezuo Pigs

Yao LI¹(), Rui JIA¹, Jie LI¹, Shuangbao GUN¹^,²^,³^,*(), Qiaoli YANG¹^,³^,*(), Longlong WANG¹, Pengxia ZHANG¹, Xiaoli GAO¹, Xiaoyu HUANG¹

1. College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
2. Gansu Modern Pig Rearing Engineering and Technology Research Center, Lanzhou 730070, China
3. Gansu Diebu Juema Pig Science and Technology Backyard, Gannan 740070, China

Received:2024-01-30 Online:2024-08-23 Published:2024-08-28
Contact: Shuangbao GUN, Qiaoli YANG E-mail:LY926111@qq.com;gunsb@gsau.edu.cn;yangql0112@163.com

摘要/Abstract

摘要：

旨在探究低温对合作猪脂肪组织形态、脂代谢相关基因表达和酶活性及AMPK/PGC-1α通路的影响。本研究选择75日龄体重相近、健康状况良好的合作猪10头，常温饲养7 d后随机分为对照组((23±2)℃)和低温组((-15±2)℃)，每组5头，试验期15 d。试验结束后采集合作猪腋下和腹股沟脂肪组织，采用HE染色观察脂肪组织形态；ELISA法检测脂代谢相关酶活性；荧光定量PCR法检测脂肪棕色化产热基因和脂代谢相关基因的表达；Western blot检测AMPK/PGC-1α通路蛋白的表达。结果发现：与对照组相比，低温处理后合作猪腋下脂肪和腹股沟脂肪呈现褐色化改变，脂肪细胞数量变多且横截面积减小；腋下和腹股沟脂肪中LPL活性，IL-6 mRNA表达量及AMPK、P-AMPK和PGC-1α蛋白表达量极显著增加(P＜0.01)，LEP mRNA表达量及FAS活性极显著降低(P＜0.01)；腋下脂肪中UCP3、HSP70 mRNA表达量显著增加(P＜0.05)，PPARγ mRNA表达量显著降低(P＜0.05)，AMPK及TNF-α mRNA表达量极显著增加(P＜0.01)；腹股沟脂肪中AMPK及TNF-α mRNA表达量显著增加(P＜0.05)，UCP3及HSP70 mRNA表达量极显著增加(P＜0.01)，PPARγ mRNA表达量极显著降低(P＜0.01)；低温处理后合作猪腹股沟脂肪中PGC-1α、ADPN、FAT mRNA表达量和HSL活性极显著增加(P＜0.01)，但在腋下脂肪中变化不显著(P＞0.05)。综上，低温下合作猪腋下脂肪和腹股沟脂肪发生褐色化改变，且脂肪细胞数量增多，面积减小，机体产热增加，脂代谢关键基因LEP、ADPN、PPARγ、FAT及AMPK/PGC-1α通路可能参与机体脂质代谢。

关键词: 低温, 脂肪组织, 脂代谢, AMPK/PGC-1α

Abstract:

This study aimed to investigate the effects of low temperature on adipose tissue morphology, lipid metabolism-related gene expression and enzyme activities, and AMPK/PGC-1α pathway in Hezuo pigs. Ten Hezuo pigs with similar body weight and good health condition at 75 days of age were selected and randomly divided into a control group ((23±2)℃) and a low-temperature group ((-15±2)℃), each with 5 pigs, after being reared for 7 d at room temperature, for 15 d of the experimental period. At the end of the experiment, the adipose tissue was collected from the axilla and groin of the Hezuo pigs, and the morphology of the adipose tissue was observed by HE staining; the activity of lipid metabolism-related enzymes was detected by ELISA; the expression of adipose browning heat-producing genes and lipid metabolism-related genes was detected by fluorescence quantitative PCR; and the expression of the AMPK/PGC-1α pathway proteins was detected by Western blot. The results showed that, compared with the control group, the axillary and inguinal fat of the Hezuo pigs showed browning changes, the number of adipocytes became larger, and the cross-sectional area was reduced after the low-temperature treatment; LPL activity, IL-6 mRNA expression and AMPK, P-AMPK and PGC-1α protein expression were highly significantly increased in axillary and inguinal fat (P < 0.01), and LEP mRNA expression and FAS activity were highly significantly decreased (P < 0.01); UCP3 and HSP70 mRNA expression were significantly increased in axillary fat (P < 0.05), and PPARγ mRNA expression was significantly decreased (P < 0.05), AMPK and TNF-α mRNA expression was highly significantly increased (P < 0.01); AMPK and TNF-α mRNA expression was significantly increased (P < 0.05) in inguinal fat, UCP3 and HSP70 mRNA expression was highly significantly increased (P < 0.01), PPARγ mRNA expression was highly significantly decreased (P < 0.01); PGC-1α, ADPN, FAT mRNA expression and HSL activity were significantly increased (P < 0.05) in inguinal fat but not in axillary fat (P>0.05) in Hezuo pigs after low-temperature treatment. In summary, the axillary fat and inguinal fat of Hezuo pigs under low temperatures underwent browning, and the area of adipocytes decreased, the number increased, the body heat production increased, and the key genes of lipid metabolism LEP, ADPN, PPARγ, FAT and the AMPK/PGC-1α pathway might be involved in the lipid metabolism of the organism.

Key words: low temperature, adipose tissue, lipid metabolism, AMPK/PGC-1α

中图分类号:

S828.2

李瑶, 贾蕊, 李杰, 滚双宝, 杨巧丽, 王龙龙, 张鹏霞, 高小莉, 黄晓宇. 低温对合作猪脂肪组织形态、脂代谢相关基因表达和酶活性及AMPK/PGC-1α通路的影响[J]. 畜牧兽医学报, 2024, 55(8): 3418-3426.

Yao LI, Rui JIA, Jie LI, Shuangbao GUN, Qiaoli YANG, Longlong WANG, Pengxia ZHANG, Xiaoli GAO, Xiaoyu HUANG. Effects of Low Temperature on Adipose Tissue Morphology, Lipid Metabolism-Related Gene Expression and Enzyme Activities, and AMPK/PGC-1α Pathway in Hezuo Pigs[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3418-3426.

图/表 5

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基因 Gene	引物序列(5′-3′) Primers sequence	产物长度/bp Product length	登录号 Accession No.
UCP3	F: TCACCTTCAGGACACGTTCG R: AGGCATCCATCCTAGTGGGT	84	NM_214049.1
PGC-1α	F: CATGTGCAACCAGGACTCTGT R: GCGTCTCTGTGAGAACTGCTA	269	NM_213963.2
HSP70	F: GCCGAGAAGGACGAGTTTGA R: TGGTACAGTCCGCTGATGATG	77	NM_213766.1
FAT	F: TGGTTTAGGAATCCCACTGCC R: GCTTCAAGTGCTGGGTCAAAA	117	NM_005667694.3
LEP	F: ACCCTCATCAAGACGATTGTCAC R: ATCACATTTCTGCAAGGCAGACT	185	XM_021078503.1
ADPN	F: GGGTCACTGTCCCTAAC R: GTCCTGGTACTGGTCGT	209	NM_214370.1
PPARγ	F: CTGACCAAAGCAAAGGCGAG R: CGGAGCGAAACTGACACCC	187	XM_005669788.3
AMPK	F: ACCAGGACCCTTTGGCAGTT R: GAATCAGGTGGGCTTGTTGC	100	XM_021076522.1
IL-6	F: CCTGAACCTTCCAAAAATGG R: ACCGGTGGTGATTCTCATC	88	NM_214399.1
TNF-α	F: GCACTGAGAGCATGATCCG R: AACCTCGAAGTGCAGTAGG	161	NM_214022.1
GAPDH	F: AGTATGATTCCACCCACGGC R: TACGTAGCACCAGCATCACC	139	NM_001206359.1

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低温对合作猪脂肪组织形态、脂代谢相关基因表达和酶活性及AMPK/PGC-1α通路的影响

Effects of Low Temperature on Adipose Tissue Morphology, Lipid Metabolism-Related Gene Expression and Enzyme Activities, and AMPK/PGC-1α Pathway in Hezuo Pigs

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