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Isolation and Differentiation of Adipose-Derived Stem Cells from Porcine Subcutaneous Adipose Tissues
Published on: March 31, 2016
Ssc-miR-130b Inhibits Lipid Accumulation in Porcine Intramuscular Preadipocytes by Activating Apoptosis via
Lulu Li1, Yongfang Chen1, Lijun Wang1
1College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, China.
Abstract:
MicroRNAs (miRNAs) play pivotal roles in intramuscular fat (IMF) deposition and determine animal meat quality. The study aimed to investigate the effect of miR-130b on adipogenesis in porcine intramuscular preadipocytes (PIMPA). In the study, PIMPA were isolated from longissimus dorsi muscle and cultured until 85%-90% confluence. The cells then subjected to adipogenic induction and transient transfections with miR-130b mimic, miR-130b inhibitor, and their corresponding controls (mNC and iNC). Oil red O staining was performed to detect lipid accumulation. Differentially expressed (DE) mRNAs were screened by transcriptome sequencing, and enriched functional pathways were identified through GO and KEGG analysis. The results demonstrated that numerous DE mRNAs were identified between the miR-130b mimic and mNC groups, which were mainly enriched in apoptosis, autophagy, and AMPK signaling pathways. Mechanistically, miR-130b overexpression significantly increased Bax expression, while reducing caspase-3 and Bcl-2 expression as well as Bcl-2/Bax ratio. It also, and tended to increase cleaved caspase-3/caspase-3 ratio, thereby activating apoptosis and suppressing lipid accumulation. Furthermore, these above results can be significantly reversed by miR-130b inhibition. In conclusion, our findings highlight that miR-130b inhibits lipid accumulation by activating apoptosis via Bcl-2/Bax/caspase-3 signaling pathway in PIMPA, providing a reliable molecular target for improving animal meat quality and ameliorating human obesity.
Insights
MicroRNA-130b (miR-130b) inhibits fat deposition in pig cells by activating apoptosis. This finding offers a molecular target for improving meat quality and addressing obesity.
Area of Science:
- Molecular Biology
- Animal Science
- Biochemistry
Background:
- MicroRNAs (miRNAs) are crucial regulators of intramuscular fat (IMF) deposition, impacting animal meat quality.
- Understanding specific miRNA roles, like miR-130b, is key to manipulating adipogenesis.
- Porcine intramuscular preadipocytes (PIMPA) serve as a model for studying fat development in livestock.
Purpose of the Study:
- To investigate the functional role of miR-130b in adipogenesis within porcine intramuscular preadipocytes (PIMPA).
- To elucidate the molecular mechanisms by which miR-130b influences lipid accumulation and cell fate.
- To assess the potential of miR-130b as a target for enhancing meat quality and combating obesity.
Main Methods:
- PIMPA were isolated and cultured, followed by adipogenic induction and transfection with miR-130b mimic/inhibitor or controls.
- Lipid accumulation was quantified using Oil Red O staining.
- Transcriptome sequencing identified differentially expressed mRNAs, with functional enrichment analysis performed using GO and KEGG.
Main Results:
- miR-130b overexpression significantly altered mRNA expression, enriching pathways related to apoptosis, autophagy, and AMPK signaling.
- miR-130b upregulated Bax, decreased Bcl-2 and caspase-3, and reduced the Bcl-2/Bax ratio, promoting apoptosis.
- Apoptosis activation by miR-130b suppressed lipid accumulation, effects reversed by miR-130b inhibition.
Conclusions:
- miR-130b inhibits lipid accumulation in PIMPA by inducing apoptosis through the Bcl-2/Bax/caspase-3 signaling pathway.
- This study identifies miR-130b as a key regulator of adipogenesis with implications for meat quality improvement.
- Targeting miR-130b presents a potential strategy for ameliorating obesity and enhancing livestock production.
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