miR-383-5p Regulates Rabbit Preadipocyte Proliferation and Differentiation

Abstract

Abstract MicroRNAs (miRNAs) are a class of small non-coding RNAs with the capability of modulating gene expression at the post-transcriptional level either by inhibiting messenger RNA (mRNA) translation or by promoting mRNA degradation. A large number of studies have shown that miR-383-5p plays a key role in the development of cancer cells, but the mechanism of miR-383-5p regulating adipogenesis is still poorly understood. In our study, we found that the expression of miR-383-5p in perirenal adipose tissue of rabbits aged 35 and 70 days was higher, which was significantly different from other tissues (p < 0.05). In addition, cell counting kit 8 (CCK-8), flow cytometry and 5-ethynyl-2’-deoxyuridine (EDU) experiments were used to identify the effect of miR-383-5p on the proliferation of rabbit preadipocytes. The results showed that miR-383-5p promoted the proliferation of rabbit preadipocytes. Western Blotting (WB), qRT-PCR and Oil Red O staining experiments were used to identify the effect of miR-383-5p on the differentiation of rabbit preadipocytes. After overexpression of miR-383-5p, the expression of fat differentiation markers PPARγ, FABP4, and CEBP/α were significantly higher than that of the control group, while the inhibitory group showed the opposite trend. The results of oil red o staining showed that the number of lipid droplets in the overexpression group was significantly higher than that in the control group, while the number of lipid droplets in the inhibition group was opposite. WB experiment also showed the same trend. These results showed that miR-383-5p promoted the differentiation of rabbit preadipocytes. Moreover, 1642 (721 up-regulated and 921 down-regulated) known differentially expressed (DE) genes were screened from miR-383-5p mimic groups and miR-383-5p negative control (NC) groups by transcriptome sequencing technology. Gene Ontology Resource (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enriched the pathways related to lipid metabolism. Protein catabolism, organic nitrogen compound biosynthesis, and lipid biosynthesis were significantly enriched. Finally, using bioinformatics websites to predict the discovery of RAD51AP1 as a potential target for miR-383-5p, their target relationship was demonstrated after its dual luciferase reporting experiment. Overall, our data indicate that miR-383-5p plays a positive regulatory effect on the proliferation and differentiation of rabbit preadipocytes.