MiR-23b Promotes Porcine Preadipocyte Differentiation via SESN3 and ACSL4

Abstract

Genetic improvement of pork quality is one of the hot topics in pig germplasm innovation. Backfat thickness and intramuscular fat content are important indexes of meat quality. MiRNAs are becoming recognized as a crucial regulator of adipose development. Therefore, it is crucial to understand how miR-23b regulates fat metabolism at the molecular level. In the present study, Oil Red O staining, and Western blot were used to evaluate the effect of miR-23b on the differentiation of porcine preadipocytes. Dual-luciferase reporter gene assay, pulldown, and RIP were used to reveal the mechanism of miR-23b regulating cell differentiation. The findings demonstrated that miR-23b promotes the expression of adipogenic factors and increases the content of lipid droplets, thus promoting the differentiation of preadipocytes. Further research found that miR-23b can directly bind to the 3’UTR of SESN3 to regulate adipogenic differentiation. In addition, it was speculated that miR-23b controls cell differentiation by positively regulating the expression of ACSL4 in other ways. Here, we demonstrate that miR-23b promotes the differentiation of porcine preadipocytes by targeting SESN3 and promoting the expression of ACSL4. The present study is meaningful to the improvement of pork quality and the development of animal husbandry.

CircHOMER1 inhibits porcine adipogenesis via the miR‐23b/SIRT1 axis

“…Li et al indicated that miR‐23b ameliorated nonalcoholic steatohepatitis 30 . In addition, our previous study demonstrated that miR‐23b promoted the differentiation of porcine preadipocytes by inhibiting SESN3 expression and promoting ACSL4 expression 19 . The function of miR‐23b is contrary to that of circHOMER1.…”

“…30 In addition, our previous study demonstrated that miR-23b promoted the differentiation of porcine preadipocytes by inhibiting SESN3 expression and promoting ACSL4 expression. 19 The function of miR-23b is contrary to that of circHO-MER1. In present study, we found that circHOMER1 could directly bind to miR-23b.…”

“…Bio‐miRNA pull down was performed as previously described 19 . At cell confluence of 70%, cells were transfected with the biotin‐labeled miRNA‐23b probe and negative control probe, respectively.…”

“…Dual-luciferase reporter assay was performed as previously described. 19 According to the binding sites, the wild-and mutant-type plasmids of circHOMER1 and SIRT1 were constructed with the psiCHECK-2 vector. The sequences were shown in Supplemental Material 2.…”

“…Bio-miRNA pull down was performed as previously described. 19 At cell confluence of 70%, cells were transfected with the biotin-labeled miRNA-23b probe and negative control probe, respectively. After 48 h of transfection, cells were washed twice with precooled PBS and subjected to Bio-miRNA pull down with MiRNA Pull Down Kit (BersinBio, China).…”