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

Authors

Abstract

Objective: 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.





Materials and Methods: 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.





Results: 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.





Conclusion: 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.

Keywords


1. Carocho M, Ferreira IC. A review on antioxidants, prooxidants and related controversy: natural and synthetic compounds, screening and analysis methodologies and future perspectives. Food and Chemical Toxicology 2013; 51:15-25. 2. Oswald MC, Garnham N, Sweeney ST, Landgraf M. Regulation of neuronal development and function by ROS. FEBS letters 2018; 10.1002/1873-3468.12972. 3. Alzoghaibi MA. Concepts of oxidative stress and antioxidant defense in Crohn’s disease. World Journal of Gastroenterology: WJG 2013; 19(39):6540-47. 4. Weigert A, von Knethen A, Fuhrmann D, Dehne N, Brüne B. Redox-signals and macrophage biology (for the upcoming issue of molecular aspects of medicine on signaling by reactive oxygen species). Molecular aspects of medicine 2018. 5. Carini F, Mazzola M, Rappa F, Jurjus A, Geagea AG, Al Kattar S, Bou-Assi T, Jurjus R, Damiani P, Leone A, Tomasello G. Colorectal carcinogenesis: Role of oxidative stress and antioxidants. Anticancer Research 2017; 37(9):4759-66. 6. El Asbahani A, Miladi K, Badri W, Sala M, Addi EA, Casabianca H, El Mousadik A, Hartmann D, Jilale A, Renaud FN, Elaissari A. Essential oils: from extraction to encapsulation. International Journal of Pharmaceutics 2015; 483(1-2):220-43. 7. Raut JS, Karuppayil SM. A status review on the medicinal properties of essential oils. Industrial Crops and Products 2014; 62:250-64. 8. Rodríguez J, Martín MJ, Ruiz MA, Clares B. Current encapsulation strategies for bioactive oils: From alimentary to pharmaceutical perspectives. Food Research International 2016; 83:41-59. 9. Munir M, Hanif M, Ranjha NM. Dendrimers and their applications: a review article. Pakistan Journal of Pharmaceutical Research 2016; 2(1):55-66. 10. Adams RP. Identification of essential oils by ion trap mass spectroscopy. Academic Press 2012. 11. Schmölz L, Wallert M, Lorkowski S. Optimized incubation regime for nitric oxide measurements in murine macrophages using the Griess assay. Journal of Immunological Methods 2017; 449:68-70. 12. Bognar E, Sarszegi Z, Szabo A, Debreceni B, Kalman N, Tucsek Z, Sumegi B, Gallyas Jr F. Antioxidant and anti-inflammatory effects in RAW264. 7 macrophages of malvidin, a major red wine polyphenol. PLoS One 2013; 8(6):e65355. 13. Sadeghi H, Robati Z, Saharkhiz MJ. Variability in Zataria multiflora Bioss. Essential oil of twelve populations from Fars province, Iran. Industrial Crops and Products 2015; 67:221-6. 14. Ullah MF, Shamim U, Hanif S, Azmi AS, Hadi SM. Cellular DNA breakage by soy isoflavone genistein and its methylated structural analogue biochanin A. Molecular Nutrition & Food Research 2009; 53(11):1376-85. 15. Hirst D, Robson T. Targeting nitric oxide for cancer therapy. Journal of Pharmacy and Pharmacology 2007; 59(1):3-13. 16. Bedard K, Krause KH. The NOX family of ROS-generating NADPH oxidases: physiology and pathophysiology. Physiological Reviews 2007;87(1):245-313. 17. More P, Pai K. In vitro NADH-oxidase, NADPH-oxidase and myeloperoxidase activity of macrophages after Tinospora cordifolia (guduchi) treatment. Immunopharmacology and Immunotoxicology 2012;34(3):368-72. 18. Salehi F, Behboudi H, Kavoosi G, Ardestani SK. Monitoring ZEO apoptotic potential in 2D and 3D cell cultures and associated spectroscopic evidence on mode of interaction with DNA. Scientific Reports 2017; 7(1):2553. 19. Kavoosi G, Amirghofran Z. Chemical composition, radical scavenging and anti-oxidant capacity of Ocimum Basilicum essential oil. Journal of Essential Oil Research 2017; 29(2):189-99. 20. Kavoosi G, Teixeira da Silva JA, Saharkhiz MJ. Inhibitory effects of Zataria multiflora essential oil and its main components on nitric oxide and hydrogen peroxide production in lipopolysaccharide‐stimulated macrophages. Journal of Pharmacy and Pharmacology 2012; 64(10):1491-500. 21. Karimian P, Kavoosi G, Amirghofran Z. Anti–oxidative and anti–inflammatory effects of Tagetes minuta essential oil in activated macrophages. Asian Pacific Journal of Tropical Biomedicine 2014; 4(3):219-27. 22. Kleniewska P, Piechota A, Skibska B, Gorąca A. The NADPH oxidase family and its inhibitors. Archivum Immunologiae et Therapiae Experimentalis 2012;60(4):277-94. 23. Sarna LK, Wu N, Hwang SY, Siow YL, O K. Berberine inhibits NADPH oxidase mediated superoxide anion production in macrophages. Canadian journal of physiology and pharmacology 2010; 88(3):369-78. 24. Brand C, Ferrante A, Prager RH, Riley TV, Carson CF, Finlay-Jones JJ, Hart PH. The water-soluble components of the essential oil of Melaleuca alternifolia (tea tree oil) suppress the production of superoxide by human monocytes, but not neutrophils, activated in vitro. Inflammation Research 2001; 50(4):213-9. 25. Ding Y, Chen ZJ, Liu S, Che D, Vetter M, Chang CH. Inhibition of Nox‐4 activity by plumbagin, a plant‐derived bioactive naphthoquinone. Journal of Pharmacy and Pharmacology 2005; 57(1):111-6. 26. Billack B. Macrophage activation: role of toll-like receptors, nitric oxide, and nuclear factor kappa B. American Journal of Pharmaceutical Education 2006; 70(5):102. 27. Li W, Fan T, Zhang Y, Fan T, Zhou P, Niu X, He L. Houttuynia cordata Thunb. Volatile Oil Exhibited Anti‐inflammatory Effects In Vivo and Inhibited Nitric Oxide and Tumor Necrosis Factor‐α Production in LPS‐stimulated Mouse Peritoneal Macrophages In Vitro. Phytotherapy Research 2013; 27(11):1629-39. 28. Jeong MY, Lee JS, Lee JD, Kim NJ, Kim JW, Lim S. A combined extract of Cinnamomi Ramulus, Anemarrhenae Rhizoma and Alpiniae Officinari Rhizoma suppresses production of nitric oxide by inhibiting NF‐κB activation in RAW 264.7 cells. Phytotherapy Research. 2008; 22(6):772-7. 29. Seo HJ, Huh JE, Han JH, Jeong SJ, Jang J, Lee EO, Lee HJ, Lee HJ, Ahn KS, Kim SH. Polygoni Rhizoma Inhibits Inflammatory Response through Inactivation of Nuclear Factor‐kappaB and Mitogen Activated Protein Kinase Signaling Pathways in RAW264. 7 Mouse Macrophage Cells. Phytotherapy Research 2012; 26(2):239-45. 30. Jaya A, Shanthi P, Sachdanandam P. Modulation of oxidative/nitrosative stress and mitochondrial protective effect of Semecarpus anacardium in diabetic rats. Journal of Pharmacy and Pharmacology 2010; 62(4):507-13.