Abstract

Adipose tissue is one of the main organs for the energy storage and supply of organisms. Adipose deposition and metabolism are controlled by a cascade of transcription factors and epigenetic regulatory mechanisms. Previous studies have also shown that miR-106a plays a considerable role in the development of organisms. The regulatory mechanism of miR-106a on porcine preadipocytes is still not clear. In this study, preadipocytes were isolated from the neck subcutaneous deposits of 3–5-day old Chinese native Guanzhong black pigs using 5-ethynyl-20-deoxyuridine (EdU) staining and a CCK-8 assay to detect the number of proliferous cells and real-time qPCR (RT-qPCR) and western blot analysis to detect gene expression, as well as Oil Red O and BODIPY staining dye lipid droplets and flow cytometry (FCM) to detect cell cycles. We also used the double luciferase method to detect the relative luciferase activities. Upregulated miR-106a increased the number of proliferous cells and enhanced the expression of cell proliferation-related genes in porcine adipocytes. The double luciferase reporter vector confirmed that p21 was a target gene of miR-106a in the cell proliferation phase. miR-106a upregulation increased the number of lipid droplets and the expression of lipogenic genes and directly targeted BMP and activin membrane-bound inhibitor (BAMBI) in the process of differentiation. Our results indicated that miR-106a promotes porcine preadipocyte proliferation and differentiation by targeting p21 and BAMBI.

Highlights

  • Adipose tissue is an important endocrine organ of the body, which affects the health of the body, and affects the meat quality of livestock

  • Our findings revealed a potential mechanism regulated by miR-106a, p21, or BMP and activin membrane-bound inhibitor (BAMBI) acting in the development of preadipocytes

  • The results of the flow cytometry analysis indicated that miR-106a these results suggest that upregulated miR-106a promotes the proliferation of porcine preadipocytes, led to cell-cycle acceleration, with an increase in the fraction of S-phase cells and a declining G1 indicating that miR-106a may be a positive adipogenic regulator

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Summary

Introduction

Adipose tissue is an important endocrine organ of the body, which affects the health of the body, and affects the meat quality of livestock. White adipose tissue (WAT) dysfunction underpins the association of obesity with the development of cardiovascular disease, hepatic steatosis, and cancers [1,2,3,4]. WAT mainly includes subcutaneous fat, visceral fat, and intramuscular fat. Studies have revealed that backfat thickness is highly associated with intramuscular fat content, meat color, tenderness, drip loss, cooking loss, and collagen proteins [5]. Adipogenesis is required for the maintenance of adipose tissue functions. We isolated porcine subcutaneous pre-adipocytes as experimental materials to study the molecular mechanisms of preadipocyte adipogenesis. We isolated porcine subcutaneous pre-adipocytes as experimental materials to study the molecular mechanisms of preadipocyte adipogenesis. miRNAs are small RNAs of 18–22 nt in length that degrade gene mRNA

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