The Effect of Artemisia fragrans Willd: Essential Oil on Inducible Nitric Oxide Synthase Gene Expression and Nitric Oxide Production in Lipopolysaccharide-stimulated Murine Macrophage Cell Line
Abstract
The genus Artemisia is estimated to comprise over 800 species with anti-cancer, anti-fungal, anti-oxidant and anti-inflammatory properties. Artemisia fragrans (A. fragrans), a species that belongs to genus Artemisia, is rich in monoterpenes and sesquiterpenes derivatives. Due to anti-inflammatory properties of monoterpenes and sesquiterpenes, we aimed to investigate the effect of A. fragrans essential oil on mRNA expression of inducible nitric oxide synthase (iNOS) gene and nitric oxide (NO) production in Lipopolysaccharide (LPS) -stimulated RAW264.7 cell line. NO, which is synthesized by iNOS, is the main macrophage-derived inflammatory mediator. The oil obtained from the A. fragrans was prepared from aerial parts of the plant. Chemical composition of essential oil was analyzed by gas chromatography–mass spectrometry (GC/MS).The cytotoxicity of various concentrations of essential oil was evaluated by mitochondrial reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test assay. The effect of different doses (1.75-7 mg/mL) of A. fragrans oil on mRNA expression of iNOS gene and NO production in LPS-stimulated RAW 264.7 cells was assessed by real-time PCR method and Griess reagent, respectively. In GC/MS analyses of A. fragrans oil, 32 compounds were identified. The main components of the oil were camphor and 1, 8-cineole. The results demonstrated that the essential oil of A. fragrans (1.75- 7 mg/mL), in a dose-dependent manner, inhibits mRNA expression of iNOS induced by LPS in the RAW264.7 cells without cytotoxic effect even at higher doses. The results of iNOS were consistent with the results of NO production. Our preliminary results suggest the possible anti-inflammatory effect of A. fragrans. Further studies are needed to determine the full pharmacokinetics of A. fragrans activity in vivo.
1. Islam MN, Choi RJ, Jin SE, Kim YS, Ahn BR, Zhao D, et al. Mechanism of anti-inflammatory activity of umbelliferone 6-carboxylic acid isolated from Angelica decursiva. J Ethnopharmacol 2012; 144(1):175-81.
2. Park GM, Jun JG, Kim JK. XH-14, a novel danshen methoxybenzo[b]furan derivative, exhibits anti-inflammatory properties in lipopolysaccharide-treated RAW 264.7 cells. J Inflamm (Lond) 2013; 10(1):1.
3. Kou X, Qi S, Dai W, Luo L, Yin Z. Arctigenin inhibits lipopolysaccharide-induced iNOS expression in RAW264. 7 cells through suppressing JAK-STAT signal pathway. Int Immunopharmacol 2011; 11(8):1095-102.
4. Yoon W-J, Ham YM, Kim S-S, Yoo B-S, Moon J-Y, Baik JS, et al. Suppression of pro-inflammatory cytokines, iNOS, and COX-2 expression by brown algae Sargassum micracanthum in RAW 264.7 macrophages. Eur Asia J BioSci 2009; 3:130-43.
5. Hong SC, Jeong JB, Koo JS. Inhibitory effect of essential oil from Agastache rugosa against nitric oxide (NO) production induced by inducible nitric oxide synthase (iNOS) over-expression through NF-B and mitogen- activated protein kinase (MAPK) activation in lipopolysaccharide (LPS)-stimulated RAW264. 7 cells. Journal of Medicinal Plants Research 2012; 6(28):4494-500.
6. Mohamadabadi MA, Hassan ZM, Hosseini AZ, Noori S, Mahdavi M, Maroufizadeh S, et al. Study of immunomodulatory effects of arteether administrated intratumorally. Iran J Allergy Asthma Immunol 2013;12(1):57.
7. Tan X, Wang Y-L, Yang X-L, Zhang D-D. Ethyl acetate extract of Artemisia anomala S. moore displays potent anti-inflammatory effect. Evid Based Complement Alternat Med 2014; 2014:681352.
8. Yayeh T, Oh WJ, Park S-C, Kim T-H, Cho JY, Park H-J, et al. Phellinus baumii ethyl acetate extract inhibits lipopolysaccharide-induced iNOS, COX-2, and proinflammatory cytokine expression in RAW264. 7 cells. J Nat Med 2012; 66(1):49-54.
9. Abad MJ, Bedoya LM, Apaza L, Bermejo P. The artemisia L. Genus: a review of bioactive essential oils. Molecules 2012; 17(3):2542-66.
10. Park EJ, Pezzuto JM, Jang KH, Nam SJ, Bucarey SA,Fenical W. Suppression of nitric oxide synthase by thienodolin in lipopolysaccharide-stimulated RAW 264.7 murine macrophage cells. Nat Prod Commun 2012;7(6):789-94.
11. Rabiei M, Jalili A, Sefidkon F. Chemical composition of the essential oil of four Artemisia species from north of Iran. PAJOUHESH-VA-SAZANDEGI 2004.
12. Bremer K, Humphries CJ. Generic monograph of the Asteraceae-Anthemideae. Bulletin of the Natural History Museum Botany series 1993; 23(2):71-177.
13. Ackernecht E. Therapeutics From the Primitives to the 20th Century. Hafner Press, New York; 1973.
14. Asghari M, Naghavi MR, Hosseinzadeh AH, Ranjbar M, Poorebrahim M. Sequence characterized amplified region marker as a tool for selection of high-artemisinin containing species of Artemisia. Res Pharm Sci 2015;10(5):453-9.
15. Safaei-Ghomi J, Ahmadi T, Batooli H, Kashi FJ.Antioxidant and antimicrobial activity of Artemisia fragrans Willd essential oil and methanol extracts. Chemija 2012; 23(2):100-7.
16. Tang R, Sun J-L, Yin J, Li Z. Artemisia allergy research in China. Biomed Res Int 2015; 2015:179426.
17. Babaahmadi M, Amjad L, Roozbehani S. The effect of allergenicity of Artemisia aucheri flowering taps in guinea pigs. International Journal of Agriculture and Crop Sciences 2013; 5(18):2079.
18. Leng X, Ye ST. An investigation on in vivo allergenicity of Artemisia annua leaves and stems. Asian Pac J Allergy Immunol 1987; 5(2):125-8.
19. Rustaiyan A, Faridchehr A. A review on constituents and biological activities of further Iranian Artemisia Species.
20. de Vries PJ, Dien TK. Clinical pharmacology and therapeutic potential of artemisinin and its derivatives in the treatment of malaria. Drugs 1996; 52(6):818-36.
21. Yoon WJ, Moon JY, Song G, Lee YK, Han MS, Lee JS, et al. Artemisia fukudo essential oil attenuates LPS- induced inflammation by suppressing NF-kappaB and MAPK activation in RAW 264.7 macrophages. Food Chem Toxicol 2010; 48(5):1222-9.
22. Ryu JC, Park SM, Hwangbo M, Byun SH, Ku SK, Kim YW, et al. Methanol Extract of Artemisia apiacea Hance Attenuates the Expression of Inflammatory Mediators via NF- kappa B Inactivation. Evid Based Complement Alternat Med 2013; 2013:494681.
23. Lim H-K, Cho SK, Park S, Cho M. Inhibitory effects of an ethanol extract of Artemisia capillaris on inflammatory mediators from LPS-stimulated RAW 264.7 cells. Journal of the Korean Society for Applied Biological Chemistry 2010; 53(3):275-82.
24. Ha H, Lee H, Seo C-S, Lim H-S, Lee M-Y, Lee JK, et al.Antiatopic Dermatitis Effect of Artemisia iwayomogi in Dust Mice Extract-Sensitized Nc/Nga Mice. Evid Based Complement Alternat Med 2014; 2014:673286.
25. Einollah V, Behboud J, Jalil D-S, Vahid NB. Evaluating antibacterial activity from essential oil of Artemisia fragrans Willd. in North-Western of Iran. African Journal of Microbiology Research 2012; 6(4):834-7.
26. Barazandeh M. Essential oil composition of Artemisia fragrans Willd. from Iran. Journal of Essential Oil Research 2003;15(6):414-5.
27. Morteza-Semnani K, Akbarzadeh M, Moshiri K.Essential oil composition of Artemisia fragrans Willd. from Iran. Flavour and Fragrance Journal 2005;20(3):330-1.
28. Delazar A, Naseri M, Nahar L, Moghadam SB, Esnaashari S, Nazemiyeh H, et al. GC-MS analysis and antioxidant activities of essential oils of two cultivated Artemisia species. Chemistry of Natural Compounds 43(1):112-4.
29. de Cássia da Silveira e Sá R, Andrade LN, de Sousa DP.A review on anti-inflammatory activity of monoterpenes. Molecules 2013; 18(1):1227-54.
30. Souza S, Trindade M, Almeida JRGdS, Souza Araujo AA, Duarte MC, Gelain DP, et al. Structure–Activity Relationship of Terpenes with Anti‐Inflammatory Profile–A Systematic Review. Basic Clin Pharmacol Toxicol 2014; 115(3):244-56.
31. Silva-Filho SE, de Souza Silva-Comar FM, Wiirzler LAM, do Pinho RJ, Grespan R, Bersani-Amado CA, et al. Effect of Camphor on the Behavior of Leukocytes In vitro and In vivo in Acute Inflammatory Response. Tropical Journal of Pharmaceutical Research 2015; 13(12):2031-7.
32. Santos F, Rao V. Antiinflammatory and antinociceptive effects of 1, 8-cineole a terpenoid oxide present in many plant essential oils. Phytother Res 2000; 14(4):240-4.
33. Adams RP. Identification of essential oil components by gas chromatography/mass spectrometry: Allured publishing corporation; 2007.
34. Cho HJ, Seon MR, Lee YM, Kim J, Kim J-K, Kim SG, et al. 3, 3′-Diindolylmethane suppresses the inflammatory response to lipopolysaccharide in murine macrophages. J Nutr 2008; 138(1):17-23.
35. Chun S-C, Jee SY, Lee SG, Park SJ, Lee JR, Kim SC.Anti-inflammatory activity of the methanol extract of Moutan Cortex in LPS-activated Raw264. 7 cells. Evid Based Complement Alternat Med 2007; 4(3):327-33.
36. Kim Y, Zhao R, Park S, Lee J, Cho I, Yang C, et al.Anti‐inflammatory effects of liquiritigenin as a consequence of the inhibition of NF‐κB‐dependent iNOS and proinflammatory cytokines production. Br J Pharmacol 2008; 154(1):165-73.
37. Berridge MV, Herst PM, Tan AS. Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction. Biotechnol Annu Rev 2005; 11:127-52.
38. Joo T, Sowndhararajan K, Hong S, Lee J, Park S-Y, Kim S, et al. Inhibition of nitric oxide production in LPS- stimulated RAW 264.7 cells by stem bark of Ulmus pumila L. Saudi J Biol Sci 2014; 21(5):427-35.
39. Miguel MG. Antioxidant and anti-inflammatory activities of essential oils: a short review. Molecules 2010;15(12):9252-87.
40. Kumar M, Kumar S, Kaur S. Role of ROS and COX-2/iNOS inhibition in cancer chemoprevention: a review. Phytochemistry Reviews 2012; 11(2-3):309-37.
41. Ayele Y, Kim JA, Park E, Kim YJ, Retta N, Dessie G, Rhee SK, et al. A methanol extract of Adansonia digitata L. leaves inhibits pro-inflammatory iNOS possibly via the inhibition of NF-κB activation 2013;21(2):146-52.
42. Yoon JH, Baek SJ. Molecular targets of dietary polyphenols with anti-inflammatory properties. Yonsei Med J 2005; 46(5):585-96.
43. Agnihotri S, Wakode S, Agnihotri A. An overview on anti-inflammatory properties and chemo-profiles of plants used in traditional medicine. Indian journal of natural products and resources 2010; 1(2):150-67.
44. Surh YJ, Chun KS, Cha HH, Han SS, Keum YS, Park KK, et al. Molecular mechanisms underlying chemopreventive activities of anti-inflammatory phytochemicals: down-regulation of COX-2 and iNOS through suppression of NF-kappa B activation. Mutat Res2001; 480-481:243-68.
45. Szliszka E, Skaba D, Czuba ZP, Krol W. Inhibition of Inflammatory Mediators by Neobavaisoflavone in Activated RAW264.7 Macrophages. Molecules 2011;16(5):3701-12.
46. Murakami A, Ohigashi H. Targeting NOX, INOS and COX-2 in inflammatory cells: chemoprevention using food phytochemicals. Int J Cancer 2007; 121(11):2357-63.
47. Dudhgaonkar S, Thyagarajan A, Sliva D. Suppression of the inflammatory response by triterpenes isolated from the mushroom Ganoderma lucidum. Int Immunopharmacol 2009; 9(11):1272-80.
48. Bellik Y, Boukraâ L, Alzahrani HA, Bakhotmah BA, Abdellah F, Hammoudi SM, et al. Molecular mechanism underlying anti-inflammatory and anti-allergic activities of phytochemicals: an update. Molecules 2012;18(1):322-53.
49. Pan M-H, Chiou Y-S, Tsai M-L, Ho C-T. Anti- inflammatory activity of traditional Chinese medicinal herbs. J Tradit Complement Med 2011; 1(1):8-24.
50. Rathee P, Chaudhary H, Rathee S, Rathee D, Kumar V, Kohli K. Mechanism of action of flavonoids as anti- inflammatory agents: a review. Inflamm Allergy Drug Targets 2009: (3):229-35
51. Pi J, Li T, Liu J, Su X, Wang R, Yang F, et al. Detection of lipopolysaccharide induced inflammatory responses in RAW264.7 macrophages using atomic force microscope. Micron 2014; 65:1-9.
52. Bak MJ, Truong VL, Kang HS, Jun M, Jeong WS. Anti- inflammatory effect of procyanidins from wild grape (Vitis amurensis) seeds in LPS-induced RAW 264.7 cells. Oxid Med Cell Longev 2013; 2013:409321.
53. Maeda H, Yamazaki M, Katagata Y. Kuromoji (Lindera umbellata) essential oil inhibits LPS-induced inflammation in RAW 264.7 cells. Biosci Biotechnol Biochem 2013; 77(3):482-6.
54. Shafaghat A, Noormohammadi Y, Zaifizadeh M.Composition and antibacterial activity of essential oils of Artemisia fragrans Willd. leaves and roots from Iran. Nat Prod Commun 2009; 4(2):279-82.
55. Juteau F, Masotti V, Bessiere JM, Dherbomez M, Viano J. Antibacterial and antioxidant activities of Artemisia annua essential oil. Fitoterapia. 2002;73(6):532-5.
56. Khazraei-Alizadeh K, Rustaiyan A. Composition of the volatile oil of Artemisia diffusa Krasch. ex Poljak. growing wild in Iran. Journal of Essential Oil Research. 2001;13(3):185-6.
57. Heravi MJ, Sereshti H. Determination of essential oil components of Artemisia haussknechtii Boiss. using simultaneous hydrodistillation-static headspace liquid phase microextraction-gas chromatography mass spectrometry. Journal of Chromatography A.2007;1160(1):81-9.
58. Lopes-Lutz D, Alviano DS, Alviano CS, Kolodziejczyk PP. Screening of chemical composition, antimicrobial and antioxidant activities of Artemisia essential oils. Phytochemistry. 2008;69(8):1732-8.
59. Kordali S, Cakir A, Mavi A, Kilic H, Yildirim A.Screening of chemical composition and antifungal and antioxidant activities of the essential oils from three Turkish Artemisia species. Journal of agricultural and food chemistry. 2005;53(5):1408-16.
60. Karimian P, Kavoosi G, Amirghofran Z. Anti–oxidative and anti–inflammatory effects of Tagetes minuta essential oil in activated macrophages. Asian Pac J Trop Biomed 2014; 4(3):219-27.
61. Jeong D, Yi Y-S, Sung G-H, Yang WS, Park JG, Yoon K, et al. Anti-inflammatory activities and mechanisms of Artemisia asiatica ethanol extract. J Ethnopharmacol 2014; 152(3):487-96.
62. Zhu XX, Yang L, Li YJ, Zhang D, Chen Y, Kostecka P, et al. Effects of sesquiterpene, flavonoid and coumarin types of compounds from Artemisia annua L. on production of mediatorsof angiogenesis. Pharmacol Rep 2013; 65(2):410-20
63. Chou S-T, Peng H-Y, Hsu J-C, Lin C-C, Shih Y. Achillea millefolium L. essential oil inhibits LPS-induced oxidative stress and nitric oxide production in RAW 264.7 macrophages. Int J Mol Sci 2013; 14(7):12978-93
64. Abdel-Aziz M, Abass A, Zalata K, Al-Galel TA, Allam U, Karrouf G. Effect of dexamethasone and Nigella sativa on inducible nitric oxide synthase in the lungs of a murine model of allergic asthma. Iran J Allergy Asthma Immunol 2014; 13(5):324-34.
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Issue | Vol 15, No 6 (2016) | |
Section | Original Article(s) | |
Keywords | ||
Artemisia fragrans Essential oils Inflammation Macroghage Gene expression Nitric Oxide synthase |
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