FTO基因对鸡肌内脂肪细胞脂沉积的调控作用

doi:10.3864/j.issn.0578-1752.2025.22.017

Abstract

Abstract:

【Objective】 The fat mass and obesity-associated gene (FTO) is closely related to fat deposition in humans and livestock/poultry. However, its role in intramuscular fat deposition in chickens has not been reported yet. This study aims to clarify the effect of FTO on the proliferation and lipid droplet deposition of chicken intramuscular adipocytes, screen the key pathways and genes responsive to its regulation, and provide a theoretical basis for analyzing the molecular regulatory network of intramuscular fat deposition in chickens. 【Method】In this study, an FTO lentiviral expression vector was constructed, and chicken primary intramuscular adipocytes were transfected with the FTO lentiviral expression vector and a negative control vector. Fluorescence was observed using a fluorescence inverted microscope, and quantitative PCR (QPCR) was performed to detect changes in FTO gene expression in the transfection and negative control groups, thus ensuring successful transfection of the FTO lentiviral expression vector into the cells. Cell proliferation changes in the negative control and transfection groups were then detected using a CCK8 assay at 72 h and 96 h post-transfection with the FTO lentiviral expression vector. At the sixth day of transfection, oil red O staining was performed, lipid droplets were extracted with isopropanol to detect changes in intramuscular adipocyte lipid droplet deposition, and a glycerol assay kit was used to detect changes in glycerol content in the culture medium. Finally, key pathways and genes responding to FTO regulation of intramuscular adipocyte lipid deposition were screened through transcriptome sequencing, bioinformatics analysis, and PPI analysis. 【Result】Following transfection of intramuscular adipocytes with the FTO lentiviral expression vector, QPCR analysis revealed a significant increase in FTO mRNA expression in the transfected group compared with the control group (P<0.01), confirming successful transfection of the FTO lentiviral expression vector into the cells. The proliferation capacity of intramuscular adipocytes was significantly reduced at 72 h and 96 h post-transfection with the FTO lentiviral expression vector, compared with the control group (P<0.05). After six days of transfecting intramuscular adipocytes with the FTO lentiviral expression vector, the transfected group exhibited a significantly higher capacity for lipid droplet deposition than the negative control group (P<0.05), while the glycerol content in the culture medium was significantly lower (P<0.05). Transcriptome sequencing identified a total of 164 differentially expressed genes responding to FTO-regulated intramuscular fat deposition, with 71 genes upregulated and 93 downregulated. KEGG enrichment analysis of these differentially expressed genes revealed significant enrichment in 13 signaling pathways, including focal adhesion, regulation of actin cytoskeleton, MAPK signaling pathway, gap junction, cytokine-cytokine receptor interaction, FoxO signaling pathway, apoptosis, C-type lectin receptor signaling pathway, and TGF-beta signaling pathway (P<0.05), all of which were directly or indirectly related to lipid metabolism. Key pathways and genes responsive to FTO regulation were further screened, and PPI (Protein-Protein Interaction) analysis on the differentially expressed genes in significantly enriched pathways was performed. The results showed that PIK3R2, FGF16, FGF9, RHOA and NGF were key genes for FTO-regulated intramuscular adipocyte lipid deposition in chickens. Among them, PIK3R2, FGF16, FGF9, and RHOA were all enriched in the regulation of actin cytoskeleton pathway, which was identified as the core pathway. 【Conclusion】 The FTO gene inhibited intramuscular adipocyte proliferation, promoted lipid deposition, and suppressed lipolysis in chickens. PIK3R2, FGF16, FGF9 and RHOA (regulation of actin cytoskeleton pathway) have been identified as key genes/pathways that responded to FTO regulation of intramuscular adipocyte lipid deposition.

Key words: chicken, FTO, intramuscular fat

HUANG HuaYun, SUI YuLe, KONG Yi, LIANG Zhong, YANG MiaoMiao, LIU Xing, HAN Wei. Regulatory Effect of FTO Gene on Lipid Deposition in Chicken Intramuscular Adipocytes[J].Scientia Agricultura Sinica, 2025, 58(22): 4786-4796.

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基因 Genes	引物序列 Primer sequence	长度 Length (bp)
β-actin	F:5′-gtccaccttccagcagatgt-3′	169
β-actin	R:5′-ataaagccatgccaatctcg-3′	169
FTO	F:5′-ctggtcctccaagaagttcg-3′	159
FTO	R:5′-ctgctcttctggcaagctct-3′	159

通路 Pathway	P值P value	基因 Genes
MAPK 信号通路 MAPK signaling pathway	0.014	HGF，FGF9，PDGFB，NGF，AKT3，FGF16
细胞因子 - 细胞因子受体相互作用Cytokine-cytokine receptor interaction	0.018
叉头盒蛋白 O 信号通路 FoxO signaling pathway	0.014	CDKN2B，CDKN2A，PIK3R2，AKT3，FOXO4
转化生长因子 -β 信号通路 TGF-beta signaling pathway	0.031	CDKN2B，CDKN2A，SMAD6，RHOA
黏着斑信号通路 Focal adhesion pathway	0.003	HGF，ITGA1，RHOA，PDGFB，PIK3R2，AKT3
肌动蛋白细胞骨架的调控 Regulation of actin cytoskeleton	0.003	ITGA1，FGF9，RHOA，PDGFB，PIK3R2，FGF16
间隙连接Gap junction	0.001	TUBA4A，HTR2B，TUBB4B，PDGFB，GJA1
细胞凋亡Apoptosis	0.016	TUBA4A，NGF，PIK3R2，AKT3
Toll 样受体信号通路 Toll-like receptor signaling pathway	0.026	TLR5，PIK3R2，AKT3
C 型凝集素受体信号通路 C-type lectin receptor signaling pathway	0.029	RHOA，PIK3R2，AKT3
沙门氏感染Salmonella infection	0.002	TLR5，IFNGR2，IL18，RHOG
甲型流感Influenza A	0.019	IFNGR2，IL18，PIK3R2，AKT3
糖尿病并发症中的 AGE-RAGE 信号通路 AGE-RAGE signaling pathway in diabetic complications	0.031	PIM1，PIK3R2，AKT3

Regulatory Effect of FTO Gene on Lipid Deposition in Chicken Intramuscular Adipocytes

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