Docosahexaenoic acid attenuates LPS-stimulated inflammatory response by regulating the PPARγ/NF-κB pathways in primary bovine mammary epithelial cells.

BACKGROUND

Docosahexaenoic acid (DHA) is a major dietary n-3 polyunsaturated fatty acid (n-3 PUFA) in fish oil, and has been reported to possess a number of biological properties, such as anti-inflammatory, antitumor and immune-regulatory properties. However, whether DHA exert anti-inflammatory effect on lipopolysaccharide (LPS)-induced mastitis remains unclear. In this study, we investigate the effect and underlying mechanisms of the effects of DHA on LPS-stimulated primary bovine mammary epithelial cells (bMEC).

METHODS

The experiment was divided into six groups as followed: control group, GW9662+LPS+DHA (100μM) group, LPS and LPS+DHA (25, 50 and 100μM) groups. bMEC were treated with DHA for 3h before LPS (200μg/ml) stimulation, and incubated with the PPARγ inhibitor GW9662 for 12h before DHA treatment. The mRNA levels of TNF-α, IL-6 and IL-1β were measured by quantitative real-time PCR (qRT-PCR). Western blot was employed for measuring the transcriptional activity of NF-κB and PPARγ.

RESULTS

Our results showed that DHA pretreatment significantly decreased the mRNA expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in bMEC stimulated with LPS. Besides, DHA suppressed the phosphorylation of nuclear transcription factor-kappaB (NF-κB) p65 and degradation inhibitor of NF-κBα (IκBα) in NF-κB signal pathway, and activated proliferator activated receptor gamma (PPARγ). But, all those effects were obviously abolished by addition of GW9662, a specific inhibitor of PPARγ.

CONCLUSION

In conclusion, these results indicated that DHA may attenuate LPS-stimulated inflammatory response in bMEC by suppressing NF-κB activation through a mechanism partly dependent on PPARγ activation.