ABSTRACT
LIF is crucial in regulating embryo implantation, while HOXA10 is a marker gene for uterine receptivity. However, the specific mechanism of LIF regulating HOXA10 during cow embryo implantation has not been fully understood. To address this knowledge gap, the experiment involved treating bovine endometrial epithelial cells (BEECs) with LIF to investigate the relationship between LIF, miRNA, and HOXA10. The experimental findings revealed that applying LIF resulted in a substantial increase in the proliferation of endometrial epithelial cells. Moreover, the expressions of PI3K, AKT, HOXA10, CDK4, cyclinD1, and cyclinE1 were significantly elevated. Conversely, the expression of p21Cipl was significantly reduced. In the group that received a combination of LIF and a STAT3 inhibitor, the expression of PI3K/AKT remained significantly increased, but there was no significant change in the expression of HOXA10. When miRNA-27a-3p was overexpressed, it resulted in a decrease in both the RNA and protein expression of HOXA10. Conversely, inhibiting miRNA-27a-3p increased the RNA and protein expression of HOXA10. In the presence of LIF treatment, the expression of miRNA-27a-3p was reduced, while the expression of HOXA10 was increased. However, when LIF and a STAT3 inhibitor were combined, there was no significant change in the expression of miRNA-27a-3p or HOXA10. Consequently, LIF facilitated cell proliferation by activating the PI3K/AKT pathway. LIF controlled the expression of miRNA-27a-3p and HOXA10 in endometrial epithelial cells through STAT3, with miRNA-27a-3p negatively regulating the expression of HOXA10.
Subject(s)
MicroRNAs , Phosphatidylinositol 3-Kinases , Female , Cattle , Animals , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Endometrium/metabolism , MicroRNAs/genetics , MicroRNAs/metabolism , Epithelial Cells/physiologyABSTRACT
The objective of this study was to use luteinizing hormone-releasing hormone A3 (LRH-A3) and human chorionic gonadotrophin (HCG) to improve pregnancy rate of dairy cows during timed artificial insemination (TAI). In experiment 1, the TAI process (0 d, GnRH, 100 µg; 7 d, PGF2α, 0.4 mg; 56 hr, GnRH, 100 µg; 16 hr, AI) was applied to 160 dairy cows on 50th and 60th days after parturition respectively. In experiment 2, 320 postpartum dairy cows were treated with TAI (Group A), TAI + 25 µg LRH-A3 (Group B), TAI + 1,500 IU HCG 5 days after AI (Group C), and TAI + 25 µg LRH-A3 + 1,500 IU HCG 5 days after AI (Group D). In experiment 3, endometrial cells were treated with HCG. The results showed that TAI did not affect the pregnancy rate, while LRH-A3 and HCG increased the pregnancy rate of the cow. HCG of 5 IU/ml and 10 IU/ml increased the expressions of leukemia inhibitory factor but decreased those of interleukin-6, epidermal growth factor and vascular endothelial growth factor in endometrial cells. This study provided a plan for the use of LRH-A3 and HCG to increase pregnancy rate during TAI in dairy cows.
Subject(s)
Cattle/physiology , Chorionic Gonadotropin/pharmacology , Gonadotropin-Releasing Hormone/pharmacology , Insemination, Artificial/methods , Pregnancy, Animal/drug effects , Animals , Chorionic Gonadotropin/administration & dosage , Dairying , Endometrium/metabolism , Epidermal Growth Factor/metabolism , Female , Gonadotropin-Releasing Hormone/administration & dosage , Interleukin-6/metabolism , Leukemia Inhibitory Factor/metabolism , Pregnancy , Stimulation, Chemical , Vascular Endothelial Growth Factor A/metabolismABSTRACT
This study investigated the pharmacological inhibition of the toll-like receptor 4 (TLR4) genes as a measure to attenuate microcystin-LR (MC-LR) reproductive toxicity. Bovine Sertoli cells were pretreated with TLR4-IN-C34 (C34) for 1 hour. Thereafter the pretreated and non-pretreated Sertoli cells were cultured in medium containing 10% heat-activated fetal bovine serum + 80 µg/L MC-LR for 24 hours to assess the ability of TLR4-IN-C34 to attenuate the toxic effects of MC-LR. The results showed that TLR4-IN-C34 inhibited MC-LR-induced mitochondria membrane damage, mitophagy and downregulation of blood-testis barrier constituent proteins via TLR4/nuclear factor-kappaB and mitochondria-mediated apoptosis signaling pathway blockage. The downregulation of the mitochondria electron transport chain, energy production and DNA replication related genes (mt-ND2, COX-1, COX-2, ACAT, mtTFA) and upregulation of inflammatory cytokines (interleukin [IL]-6, tumor necrosis factor-α, IL-1ß, interferon-γ, IL-4, IL-10, IL-13 and transforming growth factor ß1) were modulated by TLR4-IN-C34. Taken together, we conclude that TLR4-IN-C34 inhibits MC-LR-related disruption of mitochondria membrane, mitophagy and downregulation of blood-testis barrier proteins of the bovine Sertoli cell via cytochrome c release and TLR4 signaling blockage.
