Effect Of MIR-365-3P On The Differentiation Of Precursor Adipocytes In Yanbian Yellow Cattle

MicroRNAs are noncoding RNA molecules that have a length of about 23 nucleotides and are found in viruses,plants,protozoa,and animals.Its function is to mediate post transcriptional gene silencing.By complementary pairing with the 3’UTR region of target gene mRNA,microRNAs form a complex regulatory network that causes target gene destruction or translation inhibition,so as to regulate the expression of countless genes involved in different biological and physiological processes,including adipocyte differentiation.More and more studies have confirmed that microRNAs play an important role in the regulation of adipocyte differentiation.Therefore,this study was conducted based on the differentially expressed miR-365-3p and its target gene FKBP5 screened from the longissimus dorsi muscle of Yanbian Yellow Cattle bulls and steers by the research group in the early stage.The mechanism of miR-365-3p and FKBP5 on adipocyte differentiation of Yanbian yellow cattle was studied by cell experiment,QRT PCR and Western blot.Objective to provide.theoretical basis and basis for molecular breeding of Yanbian yellow cattle.The results are as follows:1.The overexpression of bta-miR-365-3p significantly reduced the lipid accumulation and triglyceride content in adipocytes(p<0.05).The expression of lipogenic key genes C/EBP α and PPAR γ was also significantly reduced(p<0.05).The experimental results obtained by inhibiting the expression of bta-mir-365-3p are contrary to the above.This suggests that bta-miR-365-3p may regulate the adipogenic differentiation of Yanbian Yellow Cattle precursor adipocytes.2.The over expression of gene FKBP5 significantly increased the expression of lipid forming key genes C/EBP α and PPAR γ(p<0.05).Inhibition of FKBP5 decreased the expression of C/EBP α and PPAR γ.This suggests that gene FKBP5 may regulate the adipogenic differentiation of Yanbian Yellow Cattle precursor adipocytes.3.In the double Luciferase Report test,the fluorescence activity of the cotransfection group of bta-miR-365-3p mimics and Psicheck2-FKBP5-WT decreased significantly(p<0.05).It indicates that there is a targeting relationship between bta-miR-365-3p and FKBP5.Overexpression of bta-miR-365-3p significantly decreased the expression of FKBP5(p<0.05).Inhibiting the expression of bta-miR-365-3p was the opposite.It shows that bta-miR-365-3p negatively regulates FKBP5.4.Overexpression of bta-mir-365-3p significantly inhibited the expression of mRNA and protein on genes AMPK α、mTOR and SREBP1(p<0.05).Inhibiting the expression of bta-miR-365-3p was the opposite.Overexpression of FKBP5 significantly increased the expression of genes AMPK α,mTOR and SREBPI(p<0.05).Inhibiting the expression of FKBP5 was the opposite.And bta-miR-365-3p targeted regulation of gene FKBP5.This suggests that bta-miR-365-3p may participate in AMPK/mTOR signaling pathway and regulate the differentiation of Yanbian Yellow Cattle precursor adipocytes by targeting gene FKBP5.