晋南牛SREBP1基因调控前体脂肪细胞分化的研究

doi:10.11843/j.issn.0366-6964.2024.11.019

Abstract

Abstract:

This study aimed to investigate the effect of the SREBP1 gene on the differentiation of Jinnan cattle precursor adipocytes and to explore the regulatory mechanism of this gene in the differentiation of Jinnan cattle precursor adipocytes by RNA-Seq technology. We transfected Jinnan cattle preadipocytes with an overexpression plasmid of SREBP1 gene and siRNA, and after differentiation induced by oleic acid, oil red O staining was used to observe the accumulation of lipid droplets, and triglyceride (TG) content was tested to explore the influence of SREBP1 gene on their differentiation. RT-qPCR and Western blotting (WB) were used to detect changes at mRNA and protein levels of related genes in preadipocytes overexpressing the SREBP1 gene. RNA-Seq was used to detect adipocytes that interfered with the SREBP1 gene, screened differentially expressed genes (DEGs), performed enrichment analysis of the KEGG pathway, and verified the differentially expressed genes. The potential target genes of the SREBP1 gene involved in regulating the differentiation of precursor adipocytes of Jinnan cattle were explored. Three replicates were set up for each treatment. The results showed as follows: 1) Compared with the control group, overexpression of SREBP1 gene could promote the accumulation of lipid droplets in precursor adipocytes and significantly increase the intracellular triglyceride content (P < 0.01), knockdown of SREBP1 gene could inhibit lipid droplet formation. 2) FABP4 mRNA expression significantly increased after overexpression of the SREBP1 gene (P < 0.05), the expression of FABP7 and LPL were significantly increased (P < 0.01), WB results showed that FABP4 protein expression level increased obviously; 3) A total of 227 DEGs were detected by RNA-Seq, including 67 up-regulated genes and 160 down-regulated genes. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that differentially expressed genes were enriched into PPAR signaling pathway and unsaturated fatty acid biosynthesis pathway, the mRNA expression of differentially expressed genes FABP4, LPL, and FABP7 decreased significantly after interfering with SREBP1 gene (P < 0.01), the protein expression level of FABP4 also decreased obviously. In conclusion, the SREBP1 gene can promote the differentiation of precursor adipocytes of Jinnan cattle, and overexpression of this gene can promote intracellular TG accumulation, which is most likely achieved through the regulation of the PPAR signaling pathway and unsaturated fatty acid biosynthesis pathway related to lipid differentiation. The results of this study provide a theoretical reference for exploring the regulatory mechanism of the SREBP1 gene in the differentiation of bovine precursor adipocytes.

Key words: precursor adipocytes, SREBP1, induced differentiation, RNA-Seq, PPAR

CLC Number:

S823.2

Weiyu ZHANG, Jing CHENG, Jiabao XU, Jing WANG, Xinyan TAO, Bo LI, Yawei ZHANG, Dandan ZHANG, Ning ZHANG, Zhenkai HAO, Chenbo ZHOU, Yuanqing ZHANG. Regulation of Preadipocyte Differentiation by SREBP1 Gene in Jinnan Cattle[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(11): 5003-5017.

Figures/Tables 13

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基因Gene	登录号GenBank No.	引物序列(5′→3′) Primer sequence	产物长度/bp Product size
固醇调节元件结合蛋白1 SREBP1	NM_001113302.1	F：CGACACCACCAGCATCAACCACG	119
		R：GCAGCCCATTCATCAGCCAGACC
β肌动蛋白β-actin	AY141970.1	F：GCACCACACCTTCTACAACGAGC	179
		R：CAGAGGCATACAGGGACAGCAC
脂蛋白脂肪酶LPL	NM_001075120.1	F：GCCGCAGACAGGATTACAGG	107
		R：CCAGGAATGAGGTGGCAAGT
脂肪酸结合蛋白4 FABP4	NM_174314.2	F：TGAGATTTCCTTCAAATTGGG	202
		R：TGAGATTTCCTTCAAATTGGG
脂肪酸结合蛋白7 FABP7	NM_001078162.2	F: GCTTGGTGTGGGCTTTGCC	154
		R: GGTTTCATCAAACTCTTCTCCCAG

引物名称Primer name	引物序列(5′→3′)Primer sequence
si-365-sense	CCUGAAGAUAUACCCAUCUTT
si-365-antisense	AGAUGGGUAUAUCUUCAGGTT
si-1051-sense	GCUCUUCCAUCAAUGACAATT
si-1051-antisense	UUGUCAUUGAUGGAAGAGCTT
si-3142-sense	GGAGGGUAUUCCUACAUGATT
si-3142-antisense	UCAUGUAGGAAUACCCUCCTT
si-con-sense	UUCUCCGAACGUGUCACGUTT
si-con-antisense	ACGUGACACGUUCGGAGAATT
si-con-FAM-sense	UUCUCCGAACGUGUCACGUTT
si-con-FAM-antisense	ACGUGACACGUUCGGAGAATT

基因Gene	差异倍数log₂FC
SREBP1	-2.436 97
FABP4	1.616 90
FABP7	-1.032 92
PLTP	2.365 92
FADS2	-2.587 32
LPL	-1.114 32

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Regulation of Preadipocyte Differentiation by SREBP1 Gene in Jinnan Cattle

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