Abstract
Simple SummarySeveral studies have shown that miR-193b plays an important role in preadipocyte differentiation. Herein, we explored the role of bta-miR-193b in adipocyte development, using EdU, flow cytometry, CCK-8, RT-qPCR, Western blotting, and oil red O staining. The result showed that bta-miR-193b could regulate the proliferation, differentiation and apoptosis of adipocytes. The dual-fluorescent reporter vector experiments showed that bta-miR-193b directly targeted ACSS2, and the regulatory function of ACSS2 is opposite to that of miR-193b. Meanwhile, we demonstrated that ACSS2 could significantly promote the expression of AKT and pAKT proteins. In conclusion, our research provides new insights that confirm that bta-miR-193b inhibits bovine adipose cell proliferation, and promotes apoptosis by negative regulation of the ACSS2/AKT pathway.The precise functions and molecular mechanisms of microRNAs (miRNAs) in adipocytes are primarily unknown. Studies have demonstrated that miR-193b plays a pivotal role in the differentiation of preadipocytes. Herein, we evaluated the effects of bta-miR-193b on the growth and development of adipocytes, using the EdU cell proliferation method, flow cytometry analysis, CCK-8 assay, RT-qPCR, Western blotting, and oil red O staining. We observed that the overexpression of bta-miR-193b significantly affected the differentiation, proliferation, and apoptosis of adipocytes. The results of the dual-fluorescent reporter vector experiments demonstrated that bta-miR-193b directly targeted Acyl-CoA synthetase short-chain family member 2 (ACSS2). Additionally, the effects of ACSS2 overexpression on the proliferation and apoptosis in adipose cells were the opposite of those induced by bta-miR-193b. We also demonstrated that ACSS2 can significantly promote the expression of AKT and pAKT proteins. Therefore, this study presents a novel mechanism by which bta-miR-193b regulates adipocyte development by targeting ACSS2.
Highlights
Adipose, one of the largest tissues in an animal body, is formed in the womb during the perinatal period and continues to grow throughout life
The results showed that, compared with the control group, more lipid droplet formation was detected in the pcDNA-miR-193b-transfected adipocytes by oil red O staining on day 5 of differentiation
The analysis of the expression profiles demonstrated that the expression of Acyl-CoA synthetase short-chain family member 2 (ACSS2) was highest in the adipose tissue, indicating that it plays an important role in the cattle adipose tissue (Figure 4d)
Summary
One of the largest tissues in an animal body, is formed in the womb during the perinatal period and continues to grow throughout life. Despite the regulation of the metabolic balance by adipose tissue via the secretion adipokines, excessive adipose accumulation can cause metabolic disorders, leading to obesity [2]. Previous studies have demonstrated that excessive fat deposition is associated with the Animals 2020, 10, 1265; doi:10.3390/ani10081265 www.mdpi.com/journal/animals. Animals 2020, 10, 1265 incidence of cancer, hypertension, type 2 diabetes mellitus, and other diseases [3]. Previous studies have demonstrated that many miRNAs can regulate the growth and development of mammalian adipocytes, and can have a potential effect on adipogenic differentiation disorders [5]. Elucidating the regulatory mechanisms of the growth and development of fat cells has turned out to be an important research direction for the prevention and treatment of obesity and associated metabolic diseases
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