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Knockdown of LXR? Inhibits Goat Intramuscular Preadipocyte Differentiation.

ABSTRACT: Goat intramuscular fat (IMF) content is mainly determined by the processes of intramuscular preadipocytes adipogenic differentiation and mature adipocyte lipid accumulation. However, the underlying regulators of these biological processes remain largely unknown. Here, we report that the expression of Liver X receptor alpha (LXR?) reaches a peak at early stage and then gradually decreases during goat intramuscular adipogenesis. Knockdown of LXR? mediated by two independent siRNAs significantly inhibits intramuscular adipocytes lipid accumulation and upregulates preadipocytes marker- preadipocyte factor 1 (pref1) expression. Consistently, siRNA treatments robustly decrease mRNA level of adipogenic related genes, including CCAAT enhancer binding protein alpha (Cebp?), Peroxisome proliferator activated receptor gamma (Pparg), Sterol regulatory element binding protein isoform 1c (Srebp1c), Fatty acids binding protein (aP2) and Lipoprotein lipase (Lpl). Next, adenovirus overexpression of LXR? does not affect intramuscular adipocytes adipogenesis manifested by Oil Red O signal measurement and adipogenic specific genes detection. Mechanically, we found that both CCAAT enhancer binding protein beta (Cebp?) and Kruppel like factor 8 (Klf8) are potential targets of LXR?, indicated by having putative binding sites of LXR? at the promoter of these genes and similar expression pattern during adipogenesis comparing to LXR?. Importantly, mRNA levels of Cebp? and Klf8 are downregulated significantly in goat LXR? knockdown intramuscular adipocyte. These results demonstrate that loss function of LXR? inhibits intramuscular adipogenesis possibly through down-regulation of Cebp? and Klf8. Our research will provide new insights into mechanical regulation of goat IMF deposition.

SUBMITTER: Xiong Y

PROVIDER: S-EPMC6213902 | biostudies-literature | 2018 Oct

REPOSITORIES: biostudies-literature

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Knockdown of LXRα Inhibits Goat Intramuscular Preadipocyte Differentiation.

Xiong Yan Y Xu Qing Q Lin Sen S Wang Yong Y Lin Yaqiu Y Zhu Jiangjiang J

International journal of molecular sciences 20181005 10

Goat intramuscular fat (IMF) content is mainly determined by the processes of intramuscular preadipocytes adipogenic differentiation and mature adipocyte lipid accumulation. However, the underlying regulators of these biological processes remain largely unknown. Here, we report that the expression of Liver X receptor alpha (<i>LXRα</i>) reaches a peak at early stage and then gradually decreases during goat intramuscular adipogenesis. Knockdown of LXRα mediated by two independent siRNAs significa ...[more]

PMID: 30301149

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Project description:CPT1B, as the major rate-limiting enzyme for fatty acid β-oxidation, involved in the regulation of adipose tissue lipid deposition, but its role in the regulation of goat intramuscular fat (IMF) remains unclear. Here, we conducted knockdown experiments targeting the CPT1B gene in goat intramuscular preadipocytes, resulting in a significant increase in intracellular triglyceride (TAG) content and lipid droplet accumulation. Conversely, the overexpression of CPT1B led to a significant decrease in intracellular TAG content and lipid droplet accumulation. Mechanistically, through RNA-seq analysis of CPT1B knockdown samples, we identified 285 differentially expressed genes (DEGs), including 71 up-regulated and 214 down-regulated genes. Notably, these DEGs were significantly enriched in various KEGG signaling pathways, such as fatty acid metabolism, focal adhesion, FoxO signaling, PI3K-Akt signaling, and MAPK signaling pathways. Gene Set Enrichment Analysis (GSEA) further confirmed the upregulation of the MAPK signaling pathway upon CPT1B knockdown in adipocytes. In rescue experiments, we demonstrated that the addition of a p38MAPK inhibitor (PD169316) to CPT1B-knockdown adipocytes counteracted the increase in lipid deposition caused by the loss of CPT1B function. These findings suggest that CPT1B inhibits lipid deposition in goat intramuscular preadipocytes via the p38MAPK signaling pathway. In addition, we observed a potential interaction between CPT1B and CPT1A through Protein-Protein Interaction Networks (PPIs). Interestingly, CPT1A exhibited a similar regulatory function to CPT1B, and they both were able to partially restore the changes in lipid deposition induced by each other. This observation supports the hypothesis of synergistic effects between CPT1B and CPT1A. Collectively, our results elucidate the role of CPT1B in the regulation of lipid deposition in goat intramuscular adipocytes through the p38MAPK signaling pathway, providing valuable insights for enhancing the quality of goat intramuscular fat.

Dataset Information

Knockdown of LXR? Inhibits Goat Intramuscular Preadipocyte Differentiation.

Publications

Knockdown of LXRα Inhibits Goat Intramuscular Preadipocyte Differentiation.

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