Chronic postpartum uterine
infection detrimentally affects subsequent fertility. Nonsteroidal anti-inflammatory drugs (
NSAID) are used to alleviate
pain and treat inflammatory conditions in transition dairy cows with varying success. To screen the efficacy of
NSAID in the absence of animal experiments, we have established an in vitro model to study uterine
inflammation.
Inflammation was induced in cultured bovine endometrial epithelial cells by challenging cells with an
inflammation cocktail:
lipopolysaccharide and proinflammatory
cytokines, interleukin-1β (IL1β) and
tumor necrosis factor α (TNFα). Release of the
inflammation markers, serum amyloid A (SAA) and α-1-acid
glycoprotein (αAGP), was measured by ELISA. Concentration of these markers was used to indicate the effectiveness in dampening
inflammation of 5
NSAID:
meloxicam,
flunixin meglumine,
aspirin,
ketoprofen, and
tolfenamic acid. Three
NSAID,
meloxicam,
flunixin meglumine, and
tolfenamic acid, were successful at dampening the release of SAA and αAGP into cell-culture supernatant, and the corresponding treated cells were selected for down-stream
mRNA expression analysis. Expression of 192 genes involved in regulation of inflammatory pathways were investigated using Nanostring. Of the genes investigated, 81 were above the
mRNA expression-analysis threshold criteria and were included in expression analysis. All SAA genes investigated (SAA2, SAA3, M-SAA3.2) were upregulated in response to the
inflammation cocktail, relative to
mRNA expression in control cells; however, AGP
mRNA expression was below the expression analysis threshold and was, therefore, excluded from analysis. Treatment with
NSAID downregulated genes involved in regulating
chemokine signaling (e.g., CXCL2, CXCR4, CXCL5, and CXCL16) and genes that regulate the
eicosanoid pathway (e.g., LTA4H,
PTGS2, PLA2G4A, and PTGDS). Of the 5
NSAID investigated,
meloxicam,
flunixin meglumine, and
tolfenamic acid are recommended for further investigation into treatment of postpartum uterine
inflammation. The results from this study confirm the immunomodulatory properties of the endometrial epithelium in response to inflammatory stimuli and suggest that
NSAID may be beneficial in alleviating uterine
inflammation.