ACSBG2基因通过类固醇激素合成和细胞黏附相关通路介导鹅肝组织对营养状态变化的响应

doi:10.11843/j.issn.0366-6964.2024.11.020

摘要/Abstract

摘要：

旨在探究酰基辅酶A合成酶家族成员ACSBG2基因在鹅肝组织响应营养状态变化中的作用及相关机制。本研究将10日龄健康朗德鹅(n=8)用于禁食/再饲喂模型，将65日龄朗德鹅(n=6)用于过量饲喂模型。首先，利用上述动物模型检测鹅肝及胸肌中ACSBG2的mRNA表达对营养状态变化的响应；其次体外培养鹅原代肝细胞及肌细胞，检测营养相关因子(葡萄糖、胰岛素、油酸钠、棕榈酸)对ACSBG2 mRNA表达水平的影响；然后通过在鹅原代肝细胞中过表达ACSBG2基因并进行转录组测序分析，以筛查ACSBG2调控的下游基因与通路；最后在体外培养细胞和动物模型中检测ACSBG2下游基因的mRNA表达水平。结果发现：1)禁食可抑制肝中ACSBG2的表达(P < 0.01)，再饲喂和过量饲喂可诱导肝中ACSBG2的表达(P < 0.001)；再饲喂(P < 0.001)和过量饲喂(P < 0.01)可诱导胸肌中ACSBG2的表达。2)油酸钠(P < 0.05)和棕榈酸(P < 0.001)可抑制鹅原代肝细胞中ACSBG2的表达；葡萄糖(P < 0.01)和胰岛素(P < 0.001)可诱导鹅原代肝细胞中ACSBG2的表达；葡萄糖(P < 0.05)和棕榈酸(P < 0.05)可抑制鹅原代肌细胞中ACSBG2的表达，油酸钠可诱导鹅原代肌细胞中ACSBG2的表达(P < 0.001)。3)ACSBG2过表达所影响的差异表达基因主要富集于类固醇激素合成和细胞黏附相关的通路。4)在体外和动物模型中ACSBG2可能介导了营养状态变化对STX1A基因表达的影响。本研究表明，ACSBG2基因主要通过类固醇激素合成与细胞黏附等通路调控糖脂代谢和炎症相关因子的表达，从而介导营养/能量变化所造成的影响。

关键词: 鹅, ACSBG2基因, 能量代谢, 类固醇激素, 细胞黏附

Abstract:

The aim of this study was to explore the role and related mechanism of ACSBG2 gene, a member of acyl-CoA synthetase family, in the response of goose liver to changes in nutritional status. In this study, 10-day-old healthy Landes geese (n=8) were used in the fasting/refeeding model, and 65-day-old Landes geese (n=6) were used in the overfeeding model. Firstly, these animal models were used to determine the response of ACSBG2 mRNA expression in goose liver and pectoral muscle to changes in nutritional status. Secondly, the effects of nutrition-related factors (glucose, insulin, sodium oleate, palmitic acid) on the mRNA expression level of ACSBG2 in goose primary hepatocytes and pectoral myocytes were determined. Then, ACSBG2 gene was overexpressed in goose primary hepatocytes and transcriptome sequencing analysis was performed to identify the downstream genes and pathways regulated by ACSBG2, which was followed by determining the mRNA expression level of ACSBG2 downstream genes in vitro and in animal models. The results showed that: 1) The expression of ACSBG2 in the liver was inhibited by fasting (P < 0.01) and induced by refeeding and overfeeding (P < 0.001). The expression of ACSBG2 in pectoral muscle was induced by refeeding (P < 0.001) and overfeeding (P < 0.01). 2) The expression of ACSBG2 in goose primary hepatocytes was inhibited by sodium oleate (P < 0.05) and palmitic acid (P < 0.001), but induced by glucose (P < 0.01) and insulin (P < 0.001). The expression of ACSBG2 in goose primary pectoral cells was inhibited by glucose (P < 0.05) and palmitic acid (P < 0.05), but induced by sodium oleate (P < 0.001). 3) The differentially expressed genes affected by ACSBG2 overexpression were mainly enriched in steroid hormone synthesis and cell adhesion-related pathways. 4) ACSBG2 may mediate the effect of changes in nutritional status on STX1A gene expression in vitro and animal models. In summary, ACSBG2 gene mainly regulates glycolipid metabolism and the expression of inflammation-related factors through steroid hormone synthesis and cell adhesion-related pathways, thereby mediating the effects of nutritional/energy changes.

Key words: goose, ACSBG2 gene, energy metabolism, steroid hormone, cell adhesion

中图分类号:

S835.2

王婉昕, 袁紫金, 朱功全, 王雨晴, 薛颖, 葛晶, 赵敏孟, 刘龙, 龚道清, 耿拓宇. ACSBG2基因通过类固醇激素合成和细胞黏附相关通路介导鹅肝组织对营养状态变化的响应[J]. 畜牧兽医学报, 2024, 55(11): 5018-5034.

Wanxin WANG, Zijin YUAN, Gongquan ZHU, Yuqing WANG, Ying XUE, Jing GE, Minmeng ZHAO, Long LIU, Daoqing GONG, Tuoyu GENG. ACSBG2 Gene Mediates the Response of Goose Liver to Nutritional Changes through Steroid Hormone Synthesis and Cell Adhesion-related Pathways[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(11): 5018-5034.

图/表 16

表 1

荧光定量引物序列"

基因Gene	引物序列(5′→3′) Primer sequence (5′→3′)	产物大小/bp Product size	退火温度/℃ Annealing temperature
ACSBG2	F: AGGTGGCCAGTGGATAAAGC	214	62
	R: ATGACAGGCCTCAGGAGAGT
UBC	F: AGGGTGGATTCTTTCTGG	243	55
	R: ACTGAGTTTGGAGGGAGC
CYP8B1	F: TGATGGACCCCTTCTGCTTT	227	60
	R: CGGCTTGGAAAACATCAGCT
ADIPOQ	F: CAAGTTCCTCTGCAGCATCC	134	61
	R: TGTTGGTCTGGAACTGGTCA
COL1A1	F: CAGATCGAGAACATCCGCAG	248	61
	R: TGCTTCTTCTCCTTGGGGTT
EREG	F: CTGCTCTTCCTCGGTTACCT	152	62
	R: GGCTTGCATCTGGTTATCCC
ANGPT4	F: GACAACGACAACTGCCTCTG	126	61
	R: GTTGAGCTTGCGGATGTTGT
NEU3	F: CTTCTGCCTCTTCGCCCT	176	59
	R: AATTAAAAGCCGCACGACCC
LCAT	F: TGTGGGAGGACATGAAGGAC	163	62
	R: GTGTCATCCCCATCCCCATA
HES1	F: GAGTGCATGAACGAGGTGAC	109	62
	R: CGTTGATCTGGGTCATGCAG
FBXO32	F: AAGCTCTGCCAGTACCACTT	242	60
	R: TCTTGCGGTGAAAGAGAGGT
FAM32A	F: ACGGAAGAAGAAGAAGGCGA	153	61
	R: CTCTCCATTTGCCGCTTCTC
HLA-DRA	F: CGTCCATCATCACCATCACG	116	62
	R: GGGCAGGTAGGAGAACTTGT
TMEM14A	F: CGGACTAGTAGCATGCCTGA	141	61
	R: TCCACTGAAACACTCTGCCT
RPL18A	F: CAAGTCCCGGTTCTGGTACT	92	62
	R: ACTTCTCATACACCTGGCCG
C1QA	F: AGCATCACCAAGAACAGGGA	155	61
	R: CTGTAGATCTGGCTGCCCTT
THBS2	F: AGGACAGGCAATCTAGAGGC	209	62
	R: GTAGTTCTCCCCTGCCACTT
SIGLEC1	F: CCAAAAGACCGTGGTGTACC	150	62
	R: CGGAACTTATAGGACCCGCT
SCARA3	F: TACCTCAACAAGTCCCTCGG	164	62
	R: ATGTTCCTCAGCATCTCCCC
CNTNAP1	F: CCGGCATCAACAACTACGTG	207	62
	R: CTGCGAACTCCAGCTTCTTC
CLDN3	F: GAAGGGCTGTGGATGAACTG	221	62
	R: GAGACGATGGTGATCTTGGC
STX1A	F: ATCCGAGAGCTGCATGACAT	204	60
	R: ACCAAGGATCACACAGCAGA
RPL27	F: AGCGAAGAGGTCCAAGATCA	195	60
	R: TTCTTGCCCGTCTTGTACCT
RPL18A	F: CAAGTCCCGGTTCTGGTACT	225	62
	R: GCTCCCATATCACGGTAGCA
ENSACDG00005014361	F: GAACCCCAGCTACCAAAACG	153	62
	R: CCACCAAAGGCACCTGAATG
ENSACDG00005013890	F: CTGTCCTGGTGCCTCATGTA	177	62
	R: GAAGGGCAGGAAGGAAGAGT
ACTB	F: CAACGAGCGGTTCAGGTGT	92	61
	R: TGGAGTTGAAGGTGGTCTCG

表 1

图 1

图 2

图 3

图 4

图 5

图 6

表 2

图 7

图 8

图 9

图 10

图 11

图 12

图 13

图 14

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基因表达变化Gene expression change	ID	基因Gene	倍数的对数log₂(fold change)	P_adj
上调Up-regulate	ENSACDG00005017918	ASCBG2	2.88	7.67×10^-285
	ENSACDG00005010507	-	1.20	1.73×10^-55
	ENSACDG00005010937	RPL18A	1.74	1.35×10^-27
	ENSACDG00005013321	GPNMB	1.55	2.27×10^-14
	ENSACDG00005003993	-	1.11	6.81×10^-14
	ENSACDG00005011759	-	1.01	3.85×10^-13
	ENSACDG00005010479	TMEN14A	1.03	1.39×10^-12
	ENSACDG00005012911	TMEN52	1.03	3.10×10^-11
	ENSACDG00005013890	-	1.30	3.39×10^-10
	ENSACDG00005015365	-	1.26	3.87×10^-9
下调Down-regulated	ENSACDG00005007930	-	-1.21	3.76×10^-54
	ENSACDG00005008996	C1QB	-1.55	5.37×10^-53
	ENSACDG00005009001	C1QA	-1.35	6.34×10^-50
	ENSACDG00005014182	SIGLEC1	-1.21	3.08×10^-41
	ENSACDG00005016869	-	-1.17	1.22×10^-38
	ENSACDG00005015981	COL1A1	-1.01	2.45×10^-35
	ENSACDG00005008999	C1QC	-1.47	2.62×10^-35
	ENSACDG00005015382	HLA-DRA	-1.014	2.62×10^-35
	ENSACDG00005009902	-	-1.23	2.30×10^-27
	ENSACDG00005018406	DBNL	-1.20	1.33×10^-25

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