The Regulation Of MiR-221-3p On The Differentiation Of Rabbit Precursor Adipocytes

Participating in the differentiation of adipocytes is one of the earliest discovered functions of miRNA.In order to explore the effect and role of miR-221-3p on the differentiation of rabbit precursor adipocytes,this experiment used rabbit precursor adipocytes as material,and utilized miR-221-3p mimic and inhibitor to transfected the rabbit preadipocytes,then induced the rabbit preadipocytes into differentiation and we detected the content of intracellular lipid droplets and the m RNA expression level of adipogenic genes during induction of differentiation.We predicted the potential targeting relationship between miR-221-3p and target gene.In order to further explore the effect of the SOCS3 gene on the differentiation of rabbit precursor adipocytes.All the results are as follows:(1)We successfully isolated and cultured the precursor adipocytes of rabbits.After 9days of differentiation,a large number of lipid droplets appeared in the cells.The detection of adipogenesis-related genes conformed to the differentiation rule of precursor adipocytes.The expression level on the 6th,9th and 12 th day of differentiation was significantly higher than that before the differentiation(P <0.01),and the expression level was the highest on the 6th day.(2)After transfection of miR-221-3p mimic,the expression level of miR-221-3p in the cells was significantly up-regulated(P <0.01).After induction of precursor adipocyte differentiation,the results showed that mimic can inhibit the differentiation of precursor adipocytes,and the m RNA expression levels of PPARγ and C/EBPα were significantly reduced(P <0.05).After transfection with miR-221-3p inhibitor,the expression level of miR-221-3p in cells was significantly down-regulated(P <0.01).The results showed that inhibitor could promote the differentiation of rabbit precursor adipocytes,the m RNA expression levels of PPARγ and C/EBPα were significantly increased(P <0.05).This result indicates that miR-221-3p inhibits the differentiation of rabbit precursor adipocytes.(3)Bioinformatics prediction results show that the SOCS3 gene is a potential target gene of miR-221-3p.There is a binding site in the 3’UTR region.According to the dual-luciferase system analysis,we found that miR-221-3p can directly target the SOCS3 gene,and extremely significantly down-regulated the firefly luciferase activity(P <0.01).(4)Overexpression of miR-221-3p can inhibit the expression level of SOCS3 gene by about 50%(P <0.01),while inhibition of miR-221-3p can increase the expression level of SOCS3 gene(P <0.01).(5)We used siRNA to transfect into cells to suppress the expression of SOCS3 gene,then we found the expression level of SOCS3 gene decreased by about 70%.After induction of precursor adipocyte differentiation,the results showed that the lipid droplets in the cells were significantly reduced,and the m RNA expression levels of PPARγ and C/EBPα were significantly reduced(P <0.01).The results showed that the differentiation of rabbit precursor adipocytes is inhibited.In conclusion,miR-221-3p inhibits the differentiation of rabbit precursor adipocytes,and miR-221-3p regulates the differentiation of rabbit precursor adipocytes by targeting the expression of the SOCS3 gene.