The effect of zataria essential oil on superoxide and nitric oxide production and NADH oxidase and nitric oxide synthase in macrophage cells

In this study, inhibitory effect of Zataria multiflora essential oil on superoxide and nitric oxide production and NADH oxidase (NOX) and nitric oxide synthase (NOS) expression was examined. Zataria essential oil was obtained by hydro-distillation and chemical composition was analyzed by gas chromatography-mass spectrometry (GC-MS). Superoxide and nitric oxide scavenging effects of zataria essential oil were tested by biochemical assays. Then, LPS-stimulated macrophages were treated by different concentrations of essential oil and expression of superoxide-producing enzyme and nitric oxide-producing enzyme was investigated by Real-time PCR, also superoxide anion and nitric oxide levels was examined by biochemical assays. The main components of the zataria essential oil were linalool (32.9%), carvacrol (26.7%), thymol (12.5%), p-Cymene (4.5%) and α-Pinene (2%). Zataria essential oil sequestered superoxide anion (O2-) and nitric oxide (NO) in in vitro assay at low concentrations of 70-100 µg/ml. Zataria essential oil at non-cytotoxic level strongly reduced intracellular reactive oxygen species (ROS) and nitric oxide (NO) production in LPS-stimulated macrophages. Superoxide-producing enzyme and NO producing enzyme expression in LPS-stimulated murine macrophage (cell line J774) was declined by zataria essential oil. Due to the results, the in vivo antioxidant activity of zataria essential oil may be attributed to down-production of NO and superoxide and also down-regulation of NOS and NOX expression.