Effects Of LncRNA 00046716 On Differentiation Of Porcine Preadipocytes And Lipid Deposition In Mature Adipocytes

Meat production and meat quality are extremely important traits in pork production.In terms of meat quality,the fat content is related to the tenderness,flavor and juiciness of the meat,and affects the overall carcass quality of the pig,which is an important factor affecting the economic benefits in pig production.Differentiation of preadipocytes and lipid accumulation in mature adipocytes significantly affect fat content.Previous studies in the reserch group showed that bone morphogenetic protein 2(BMP2)can significantly promote the differentiation of porcine preadipocytes.In the process,the expression of a large number of lncRNAs in preadipocytes is significantly changed,but their specific effects and the mechanism is not clear.Therefore,this study first isolated pig preadipocytes and cultured them in vitro.After adding 50 ng/ml of BMP2 during cell differentiation,high-throughput sequencing was used to screen differentially expressed lncRNA regulated by BMP2,and through bioinformatics.The lncRNA-mi RNA-m RNA interaction network was predicted and constructed.Subsequently,the dual luciferase reporter system,q RT-PCR,oil red O staining and other methods were used to determine the relationship between candidate lncRNA/mi RNA/m RNA and its effects on preadipocyte differentiation,lipid formation and deposition.Of the 132 lncRNAs that were differentially expressed by BMP2,61 were up-regulated and 71 down-regulated.The potential target gene of lncRNA is enriched in functions of tissue development,organ morphogenesis and cell differentiation.The KEGG pathway is enriched in the m TOR signaling pathway,lysine degradation,ribose metabolism,and insulin signaling pathways and MAPK signaling pathways associated with adipogenesis.However,lncRNA 00046716-mi RNA-149-FASN has a mutual regulatory relationship,which is reflected in the fact that lncRNA 00046716 may affect the expression of FASN,a key gene of precursor adipocyte differentiation,by binding to mi RNA-149.Moreover,the m RNA expression levels of lncRNA 00046716 and FASN increased first and then decreased during the differentiation of porcine preadipocytes.Inhibition of mi RNA-149 was able to up-regulate the expression levels of lncRNA 00046716 and FASN.Inhibition of lncRNA expression of 00046716 resulted in decreased m RNA and protein levels of the FASN gene,decreased differentiation of porcine progenitor adipocytes,and a significant decrease in triglyceride accumulation in mature adipocytes.In summary,lncRNA 00046716 can affect the expression of FASN by binding mi RNA-149,thereby regulating the differentiation of porcine preadipocytes and the accumulation of subsequent triglycerides in mature adipocytes.The results provide insights into the role and mechanism of lncRNA in adipogenesis and provide a theoretical basis for improving pork quality at the molecular level.