Abstract
Endometritis is one of the main diseases which harm sheep husbandry. Astragalin and chlorogenic acid (CGA) are common active ingredients of traditional Chinese medicine (TCM) with immunoprotective, antioxidant, and anti-inflammatory properties. In the present study, sheep endometrial epithelium cells (SEECs) were successfully purified and identified, and the in vitro inflammation model of SEECs induced by Escherichia coli (E. coli) was successfully established. To explore the effect of astragalin and CGA on the inflammation induced by E. coli and its potential mechanism, six groups were set up, namely, group C, M, astragalin, CGA, BAY, and STR. Cells in group C were incubated with DMEM/F12 for 6 h, while cells in group M, astragalin, CGA, BAY, and STR were incubated with DMEM/F12, astragalin, CGA, BAY, and STR for 3 h, respectively, followed by E. coli infection at a multiplicity of infection (MOI) of 1 E. coli per cell for 3 h. Subsequently, the cells and the supernatant were collected to detect the expression of genes in the toll-like receptor 4 (TLR4)/nuclear factor-kappa B (NF-κB) pathway by ELISA, qPCR, and western blot. The results showed that E. coli could induce inflammation of SEECs in vitro, while astragalin and CGA could alleviate the inflammatory response induced by E. coli via inhibiting the activation of the TLR4/NF-κB signaling pathway, which provides a theoretical and experimental foundation for preventing sheep endometritis clinically.
Highlights
With the rapid development of the Chinese economy and the improvement of living standards, the breeding industry gets more and more attention including sheep husbandry
The relative mRNA expression of IL-1β and IL-6 was significantly increased (P < 0.05) when compared with that in group C at multiplicity of infection (MOI) of 1:1, 5:1, 10:1, and 100:1. These results indicate that the MOI of E. coli at 1:1 was appropriate to establish the inflammation model of sheep endometrial epithelium cells (SEECs)
Bacteria are the main cause of endometritis, and one of the most common bacteria is E. coli [21, 22]
Summary
With the rapid development of the Chinese economy and the improvement of living standards, the breeding industry gets more and more attention including sheep husbandry. Not all endometritis can cause injury to the organism; for instance, endometritis induced by breeding is a normal physiological reaction in the mare, and it is a necessary inflammatory response for the effective removal of contaminating bacteria and excess spermatozoa introduced into the uterus. Persistent endometritis results from a response to the introduction of air, urine, semen, bacteria, fungus, and yeast into the uterus [2], with the leading cause being bacterial infection, E. coli, Staphylococcus aureus, and Streptococcus agalactiae [5, 6]. Lipopolysaccharide (LPS), a component of the cell wall in gram-negative bacteria, such as E. coli, could activate the toll-like receptor 4 (TLR4)/nuclear factor-kappa B (NF-κB) signaling pathway and lead to a strong inflammatory response. NF-κB translocates into the nucleus and activates the expression of inflammatory genes [8, 9]
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