miR-15a Targets ABAT Gene to Regulate the Differentiation of Preadipocytes of Yanbian Yellow Cattle

Abstract

BackgroundMicroRNAs (miRNAs) are small RNAs that cannot be translated into proteins. The length of a mature miRNA is approximately 20-25 nt. Neural regulation, inflammatory response, cell proliferation and apoptosis, fat metabolism, and cell differentiation, among others, have become research hotspots in recent years. In this experiment, dual-luciferase reporter gene detection, qRT-PCR, oil red O staining, western blotting, and other methods were used to select the differentially expressed miR-15a and its target gene, the 4-aminobutyrate aminotransferase (ABAT) gene, in the longissimus muscle tissues of cattle bulls and steers. By transfecting Yanbian yellow cattle precursor adipocytes with equal amounts of artificially synthesised miR-15a mimics, mimics NC, miR-15a inhibitor, and inhibitor NC and inducing differentiation, the total RNA and total protein of cells during the induced differentiation process were extracted.The effect of miR-15a on the mRNA and protein expression of adipogenesis-related genes was analysed. ResultsThe results showed that the lipid droplet content and lipid droplet size of the miR-15a mimic group were significantly lower than those of the mimics NC group, and the mRNA levels of the peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT enhancer-binding protein α (C/EBPα) genes in the miR-15a mimics group were significantly lower than those in the mimics NC group. At the same time, the protein expression of miR-15a mimic group cells was also significantly lower than that of the NC group cells. Meanwhile, the mRNA levels of the PPARγ and C/EBPα genes of miR-15a inhibitor group cells were significantly higher than those of the NC group cells, and the protein expression levels were also significantly higher than those of the NC group cells. The luciferase reporter results showed that miR-15a mimics significantly inhibited the activity of the ABAT-3'UTR dual luciferase reporter gene vector, indicating that the miR-15a and ABAT genes have a targeting effect, and that overexpressed miR-15a significantly reduced the expression of the ABAT gene in the cell, inhibited the expression of miR-15a, and significantly increased the expression of the ABAT.ConclusionsIn summary, miR-15a may target the ABAT gene to regulate the differentiation of Yanbian yellow cattle precursor adipocytes.

Inhibition Effect of Adipogenesis and Lipogenesis via Activation of AMPK in Preadipocytes Treated with Canavalia gladiata Extract

“…In addition, the number of triglycerides accumulated following preadipocyte cell differentiation was determined by the increase or decrease in SREBP-1c, the upregulating mechanism of adipogenesis-related genes or signal transmitters [22]. SREBP-1c, which regulates the expression of genes involved in fatty acid metabolism and lipid biosynthesis in the early stages of preadipocyte differentiation, has been shown to inhibit the expression of the PPAR-γ, C/EBP-α, and FAS sub-factors to reduce lipid accumulation [23].…”

“…In the previous analysis of the expression of adipogenesis and lipogenesis genes, the inhibition of triglyceride accumulation was confirmed by the increased expression of AMPK and CPT-1 and decreased expression of SREBP-1c, C/EBP-α, PPAR-γ, and FAS. Therefore, to explore the main substances contained in the CGE that inhibit preadipocyte In addition, the number of triglycerides accumulated following preadipocyte cell differentiation was determined by the increase or decrease in SREBP-1c, the upregulating mechanism of adipogenesis-related genes or signal transmitters [22]. SREBP-1c, which regulates the expression of genes involved in fatty acid metabolism and lipid biosynthesis in the early stages of preadipocyte differentiation, has been shown to inhibit the expression of the PPAR-γ, C/EBP-α, and FAS sub-factors to reduce lipid accumulation [23].…”