过表达DGAT2基因对延边牛前体脂肪细胞分化的影响

doi:10.16431/j.cnki.1671-7236.2023.08.002

Abstract

Abstract: 【Objective】 The purpose of this study was to investigate the effects of diacylglycerol acyltransferase 2 (DGAT2) on differentiation of bovine preadipocytes and its regulation of lipid metabolity-related signaling pathways.【Method】 The recombinant shuttle plasmid containing the target gene was constructed using the ADV1 shuttle plasmid,and then 293A cells were co-transfected with the recombinant shuttle plasmid and the skeleton plasmid pGP-Ad-Pac vector,and Adenovirus overexpressed vector Ad-DGAT2 was prepared.Bovine preadipocytes were infected with Ad-DGAT2 and Ad-NC (negative control group).96 h after differentiation induced by oleic acid,lipid droplet formation was observed by Oil red O staining,and the contents of triglyceride (TAG) and adiponectin (ADP) were detected.The expression of genes related to lipogenesis was detected by Real-time quantitative PCR.Finally,transcriptome sequencing was performed to screen differentially expressed genes,and GO functional annotation and KEGG pathway enrichment analysis were performed.【Result】 Overexpression of DGAT2 gene significantly increased the number of lipid droplets and the contents of TAG and ADP (P<0.05).Meanwhile,overexpression of DGAT2 could significantly up-regulate mRNA expression of DGAT1,glycerol phosphate acyltransferase 4 (GPAT4) and acylglycerol phosphate acyltransferase 4 (AGPAT4) in phosphoglycerol pathway and peroxisome roliferatoractivated receptor gamma (PPARγ),perilipin 2(PLIN2),CCAAT/enhancer binding protein α (C/EBPα) and stearoyl-COA desaturase (SCD) in lipid metabolism pathway (P＜0.05).Transcriptome sequencing results showed that a total of 110 differentially expressed genes were screened from preadipocytes infected with Ad-DGAT2 compared with Ad-NC group,among which 64 genes were up-regulated and 46 genes were down-regulated.KEGG pathway enrichment analysis showed that differentially expressed genes were mainly concentrated in the PPAR signaling pathway,glycerol phospholipid metabolism,fatty acid biosynthesis and AMPK signaling pathway.【Conclusion】 Overexpression of DGAT2 gene could promote lipid droplet formation,TAG accumulation and the expression of related adipogenic genes in Yanbian bovine adipocytes.Transcriptional sequencing screened out differentially expressed genes related to fat metabolism.The results indicated that DGAT2 gene played an important regulatory role in the differentiation of precursor adipocytes,which provided an important theoretical basis for molecular breeding of Yanbian bovine.

Key words: DGAT2 gene; triacylglycerol; preadipocytes; overexpression; transcriptome

CLC Number:

GUO Panpan, LI Qiang, LI Xiangzi, YAN Changguo, JIN Xin. Effect of Overexpression of DGAT2 Gene on Differentiation of Yanbian Bovine Preadipocytes[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(8): 3045-3055.

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Effect of Overexpression of DGAT2 Gene on Differentiation of Yanbian Bovine Preadipocytes

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