Time-Dependent Variations in Serum Nitrite, 6-Keto-Prostaglandin F1α and Thromboxane B2 Levels Induced by Lipopolysaccharide in Mice

Clinical features of certain immuno-inflammatory disorders exhibit time-dependent fluctuations, which could be related to circadian rhythmicity of proinflammatory mediator production. Many biologically active substances including nitric oxide (NO) and eicosanoids are released into the circulation in sepsis. Increased NO and eicosanoid levels have been reported to be responsible from death in septic shock. The aim of this study was to investigate the variations in the NO and eicosanoid production and mortality induced by bacterial endotoxin, lipopolysaccharide (LPS) injected either in the morning or in the evening. Experiments were performed on mice synchronised to 12 h light and 12 h dark (lights on at 09:00 h). Animals were injected intraperitoneally with LPS (10 mg/kg) at 09:00 (morning) and 21:00 h (evening) alone or in combination with aminoguanidine (NO synthase (NOS) inhibitor) (100 mg/kg) or indomethacin (cyclooxygenase (COX) inhibitor) (100 mg/kg). The serum was separated from blood samples obtained at nine different time points. Nitrite (stable product of NO), 6-keto-prostaglandin F1α (6-keto-PGF1α, stable product of prostacyclin) and thromboxane B2 (TxB2, stable product of thromboxane) concentrations in serum samples were measured. Serum nitrite levels showed a 24 h circadian rhythmicity depending on LPS injection time. Morning injection caused a peak after 15 h, while evening injection had two peaks after 9 and 18 h. The peak values obtained from morning and evening injections were significantly decreased by aminoguanidine and indomethacin. When LPS injected to mice in the morning and in the evening, it gradually increased the mortality rate within 24 h which could be abolished by aminoguanidine, but not indomethacin. Indomethacin-induced inhibition on LPS-induced nitrite levels was higher in the morning than in the evening. 6-keto-PGF1α and TxB2 levels were decreased by indomethacin when injected with LPS at both injection times, but not aminoguanidine. These results showed that there is an interaction between NO and eicosanoids, and LPS may produce different effects on NOS activity, but not eicosanoid production and mortality, depending on injection time in the experimental septic shock model in mice. Chronopharmacological manipulations of NOS and COX pathways and interactions between them could lead to novel therapeutic approaches for the treatment of septic shock.