胎衣不下奶牛miRNA-185靶向调控血管内皮生长因子A表达的研究

doi:10.16431/j.cnki.1671-7236.2024.03.003

Abstract

Abstract: 【Objective】 This experiment was aimed to detect the differential expression of miRNA-185 and vascular endothelial growth factor A (VEGFA) in the placental tissue of dairy cows in the normal expulsion group and retained fetal membranes (RFM) group through in vivo and in vitro experiments,as well as the expression distribution of VEGFA in the placental tissue of dairy cows.It verified and clarified the close correlation and targeted regulatory relationship between miRNA-185 and VEGFA and retained fetal membranes in dairy cows and provided theoretical and experimental basis for the in-depth study of the role of miRNA-185 in the occurrence of retained fetal membranes in dairy cows.【Method】 Real-time fluorescence quantitative PCR method was used to detect the gene expression of miRNA-185 and VEGFA in the placental tissue of dairy cows with normal and retained fetal membranes.Fluorescence in situ hybridization (FISH) was used to detect the specific and mRNA differential expression of VEGFA in the placental tissue of dairy cows.Dual luciferase reporter genes was used to clarify the targeted regulatory relationship between miRNA-185 and VEGFA,Real-time fluorescence quantitative PCR and Western blotting methods were used to detect the mRNA and protein differential expression of VEGFA after in vitro transfection of miRNA185 mimics,miRNA-185 inhibitor and miRNA-185 NC.【Result】 Real-time fluorescence quantitative PCR results showed that compared with dairy cows with normal removal of placenta,the expression levels of miRNA-185 and VEGFA in dairy cows with retained fetal membranes were extreme downregulation (P＜0.01).FISH result showed that VEGFA mRNA was mainly expressed in dairy cow endometrial epithelial cells.VEGFA was a targeted regulatory protein of miRNA-185.Compared with the negative control group,the mRNA and protein expression levels of VEGFA were extreme downregulation after transfection with miRNA-185 mimics (P＜0.01),while the mRNA and protein expression levels of VEGFA were extremely upregulated after transfection with miRNA-185 inhibitor (P＜0.01).【Conclusion】 The expression of miRNA-185 and VEGFA in the placental tissue of dairy cows with retained fetal membranes significantly decreased.VRGFA was mainly expressed in endometrial epithelial cells,and miRNA-185 could participate in the occurrence and development of retained fetal membranes in dairy cows by targeting and regulating the expression of VEGFA.

Key words: dairy cows; retained fetal membranes; miRNA-185; vascular endothelial growth factor A; maternal placenta

CLC Number:

S858.23

LUO Chunhai, ZHENG Chengyuan, ZHANG Menglong, YAO Weijia, LIU Jiajin, LIU Bingqi, WANG Wei, FU Shixin. Regulation of Vascular Endothelial Growth Factor A Expression by miRNA-185 in Dairy Cows with Retained Fetal Membranes[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(3): 916-925.

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Regulation of Vascular Endothelial Growth Factor A Expression by miRNA-185 in Dairy Cows with Retained Fetal Membranes

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