The Relationship Between MiRNA-185 Regulates VEGFA/STIM1 Genes And The Cattle With Retained Foetal Membranes

The miRNA-185 and predicted target protein(α-enolase, ENO1; vascular endothelial growth factor-A, VEGFA; stromal interaction molecule 1, STIM1) were researched in the primary dairy uterine epithelial cell by miRNA interfering Biologic technology. The miRNA mimics and miRNA inhibitors were transfected in the primary dairy uterine epithelial cell. The expression of miRNA-185 and the predicted target protein were detected. For analyzing function of miRNA-185 in the retained foetal membranes, and providing a scientific theory and experimental evidence of the disease pathogenesis to prevent and treat the disease.Maternal placenta tissue were collected in the three healthy Holstein dairy cows and the three retained placenta cows with similar age, gravida, weigh and milk yield, respectively. Expression of mi RNA-185, ENO1, VEGFA and STIM1 were detected by western blot and Q-PCR. The best method was evaluated in the primary of dairy uterine epithelial cell with different digestion and explant culture. After purifing and identifying the epithelial cell by pancreatin and CK18, miRNA-185 mimics, NC mimics, mi RNA-185 inhibitor and NC inhibitor were transfected into epithelial cell. Finally, the level of miRNA-185, ENO1, VEGFA and STIM1 expression were detected by western blot and Q-PCR.The result showed that expression of ENO1 in the R group was higher(p<0.05), VEGFA and STIM1 were lower(p<0.05) than the healthy Holstein dairy cows in the maternal placenta tissue. The two culture method were successful. However, the survival rate of cell was lower by the digestion culture, the tissue adherent culture had obviously advantages on the economy, efficient and survival rate. The purity of epithelial cell though identified CK18 was more than 80%. Expression of ENO1 was no differential after transfecting miRNA-185 mimics and NC mimics(p>0.05), STIM1 was up-regulated(p<0.01) and VEGFA was down-regulated(p<0.05); As before, expression of ENO1 was no differential after transfecting miRNA-185 inhibitor and NC inhibitor(p>0.05). However, STIM1 was down-regulated(p<0.01) and VEGFA was up-regulated(p<0.01).In brief, the level of VEGFA, STIM1 and ENO1 expression were significantly differential in the retained foetal membranes. Expression of VEGFA and STIM1 were regulated by mi RNA-185 in the dairy uterine epithelial cell. The result suggested that the expression of VEGFA and STIM1 were regulated by miRNA-185 influencing the retained placenta in the dairy cows.