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Emerging evidence has implicated nicotinamide adenine dinucleotide (NAD+) metabolism in various inflammatory diseases. In the study, the role of NAD+ metabolism in Complete Freund's Adjuvant (CFA)‐evoked inflammatory pain and the underlying mechanisms are investigated. The study demonstrated that CFA induced upregulation of nicotinamide phosphoribosyltransferase (NAMPT) in dorsal root ganglia (DRG) without significant changes in the spinal cord. Inhibition of NAMPT expression by intrathecal injection of NAMPT siRNA alleviated CFA‐induced pain‐like behavior, decreased NAD+ contents in DRG, and lowered poly‐(ADP‐ribose) polymerase 1 (PARP1) activity levels. These effects are all reversed by the supplement of nicotinamide mononucleotide (NMN). Inhibition of PARP1 expression by intrathecal injection of PARP1 siRNA alleviated CFA‐induced pain‐like behavior, while elevated NAD+ levels of DRG. The analgesic effect of inhibiting NAMPT/NAD+/PARP1 axis can be attributed to the downregulation of the NF‐κB/IL‐1β inflammatory pathway. Double immunofluorescence staining showed that the expression of NAMPT/NAD+/PARP1 axis is restricted to DRG neurons. In conclusion, PARP1 activation in response to CFA stimulation, fueled by NAMPT‐derived NAD+, mediates CFA‐induced inflammatory pain through NF‐κB/IL‐1β inflammatory pathway. [ABSTRACT FROM AUTHOR]

Background: The uteruses of most dairy cattle are easily infected by bacteria, especially gram-negative bacteria, following parturition. Macrophages are important cells of the immune system and play a critical role in the inflammatory response. In addition, cortisol levels become significantly increased due to the stress of parturition in dairy cattle, and cortisol is among the most widely used and effective therapies for many inflammatory diseases. In this study, we assessed the anti-inflammatory effects and potential molecular mechanisms of cortisol using a Lipopolysaccharide (LPS)-induced RAW264.7 macrophage cell line. Results: Cortisol significantly suppressed the production of prostaglandin E2 (PGE2) and decreased the gene and protein expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) in a dose-dependent manner. Moreover, cortisol inhibited the mRNA expression of pro-inflammatory cytokines including tumor necrosis factor alpha (TNFα), interleukin-1β (IL-1β), and interleukin-6 (IL-6) and decreased IL-1β secretion in an LPS-treated RAW264. 7 macrophage cell line. Moreover, we found that cortisol suppressed nuclear factor-kappa B (NF-κB) signaling in RAW264.7 macrophages stimulated with LPS. This suppression was mediated by the inhibition of IκBα degradation and NF-κB p65 phosphorylation. In addition, cortisol also suppressed the phosphorylation of mitogen-activated protein kinases (MAPK) such as extracellular signal-regulated kinase (ERK1/2), p38 MAPK, and c-Jun N-terminal kinase/stress-activated protein kinase (JNK). Conclusions: These results suggest that high cortisol levels can attenuate LPS-induced inflammatory responses in the RAW264.7 macrophage cell line by regulating the NF-κB and MAPK signaling pathways. [ABSTRACT FROM AUTHOR]

Progesterone suppresses the innate immune function of bovine endometrium, making the uterus susceptible to bacterial infection. The bovine endometrial epithelial cells (BEEC) are the first line of defense against bacteria, such as Escherichia coli (E.coli) that causes inflammation of endometrium through the recognition of lipopolysaccharide (LPS). The aim of this study was to investigate the effect of progesterone on the inflammatory response and its potential mechanism using E.coli - or LPS-induced BEEC. Concentrations of 1, 3 and 5 ng/mL progesterone were selected. The mRNA expressions of proinflammatory cytokines were determined using qPCR. The activations of NF-κB and MAPK pathways were detected by Western blot and immunofluorescence. An increase in the mRNA expression of IL-1β, IL-6, IL-8 and TNF-α was observed in BEEC treated with progesterone, LPS or E.coli alone. Progesterone inhibited the E.coli - or LPS-induced gene expression of these cytokines. Progesterone treatment alone showed little influence on NF-κB or MAPK pathway. In BEEC stimulated with E.coli or LPS, progesterone inhibited the phosphorylations of IκB, p65, ERK1/2, p38MAPK and JNK, and the translocation of p65 into the nucleus. These results suggested that progesterone has anti-inflammatory effect, which may be mediated by inhibiting NF-κB activation and MAPK phosphorylation in BEEC. • Progesterone alone increased the mRNA levels of IL-1β, IL-6, IL-8 and TNF-α in BEEC. • Progesterone inhibited the E.coli - or LPS-induced inflammatory response in BEEC. • Progesterone alone showed no effect on MAPK or NF-κB pathways in BEEC. • Progesterone inhibited the E.coli - or LPS-activated MAPK or NF-κB pathways in BEEC. [ABSTRACT FROM AUTHOR]

The bovine uterus is subject to infection after calving, which may lead to endometritis. Elevated cortisol levels have been observed in postpartum cattle. However, the role of cortisol in the inflammatory response of the uterus has not been reported. The aim of this study was to investigate the anti-inflammatory effects of cortisol on lipopolysaccharide (LPS)-induced primary bovine endometrial epithelial cells (BEECs). BEECs were treated with various concentrations of cortisol (5, 15 and 30 ng/mL) in the presence of LPS. The mRNA expression of TLR4 and proinflammatory cytokines was measured with qPCR. The activation of NF-κB and MAPK signalling pathways was detected with Western blotting and immunofluorescence. Cortisol induced the down-regulation of the mRNA expression of toll-like receptor 4 (TLR4) and proinflammatory cytokines, including interleukin (IL)-1β, IL-6, IL-8, tumour necrosis factor–α (TNF-α), cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS). Cortisol inhibited the activity of nuclear factor-κB (NF-κB) via blocking the phosphorylation and degradation of IκB. Cortisol suppressed the phosphorylation of mitogen-activated protein kinase (MAPK), including extracellular signal-regulated kinase (ERK1/2), p38MAPK and c-Jun N-terminal kinase/stress-activated protein kinase (JNK). These results demonstrated that cortisol may exert its anti-inflammatory actions by regulating NF-κB activation and MAPK phosphorylation. [ABSTRACT FROM AUTHOR]