Purpose
Endometritis is a female reproductive disease that affects the cattle industries development and microRNAs (miRNAs) play a pivotal role and critical regulators of the innate immune response in varieties of diseases. The present study intends to investigate the regulatory role of miR‐24-3p in the innate immune response involved in endometritis and evaluate its therapeutic potential.
Methods
Whole mice uteri and bovine endometrial epithelial cells (BEECs) were separately stimulated with LPS. The BEECs were also transfected with miR-24-3p mimic and negative control; si
TRAF6
and siNC; pcDNA3.1 empty and pcDNA3.1(+)
TRAF6
separately with LPS stimulation. The expression levels of miR‐24-3p and
TRAF6
were measured via quantitative real‐time polymerase chain reaction (qRT-PCR) and Western blot, respectively. LPS‐induced inflammatory response assessed by inflammatory cytokines secretion and expression via ELISA and qRT-PCR. Bioinformatics analysis and luciferase reporter assay validated the interaction between miR‐24-3p and
TRAF6
. The activation of the NF‐ĸB/MAPK pathway and p65 phosphorylation was investigated by Western blot and immunofluorescence assay, respectively.
Results
The expression of miR‐24-3p was decreased, and
TRAF6
was elevated with an increased level of pro-inflammatory cytokines in LPS‐treated BEECs and mice uterus. The overexpression of miR‐24-3p suppressed LPS‐induced secretion of inflammatory cytokines (IL‐1β, IL‐6, IL-8 and TNF-α) and deactivation of NF‐ĸB/MAPK pathways. The downregulation of
TRAF6
inhibited LPS‐induced inflammatory response in BEECs.
TRAF6
is validated as a target of miR‐24-3p, and miR‐24-3p reversed the overexpression of cloned
TRAF6
on inflammation response in BEECs.
Conclusion
Our findings demonstrate that the overexpression of miR‐24-3p attenuates endometrial inflammation and the expression of pro-inflammatory mediators via suppressing
TRAF6
. Therefore, modulating the pathogenesis of endometritis and possibly, a therapeutic potential against endometritis.