Anti-melanogenic activity of Viola odorata different extracts on B16F10 murine melanoma cells

Document Type : Original Article


1 Student Research Committee, Department of Pharmacology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Department of Pharmacognosy, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

3 Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran


Objective(s): In previous studies, antioxidant activity of Viola odorata L. has been demonstrated. In this study, we have investigated the anti-melanogenic effect of extract and fractions of the plant in B16F10 cell line.
Materials and Methods: Impact of different increasing concentrations of extract and fractions of V. odorata was evaluated on cell viability, cellular tyrosinase, melanin content and mushroom tyrosinase as well as ROS production in B16F10 murine melanoma cell line.
Results: Viola odorata had no cytotoxicity on B16F10 cells compared to control group. Kojic acid as positive control had significant decreasing effects on cellular and mushroom tyrosinase activity, melanin content and ROS production (P<0.001, for all cases). V. odorata (1-20 µg/ml) decreased all measured parameters including cellular tyrosinase and melanin content as well as ROS production and among all extract and fractions ethyl acetate fraction had the best effect (P<0.05).
Conclusion: Viola odorata had promising anti-melanogenic activity through inhibition of cellular tyrosinase activity and ROS production as well as melanin content. More basic and clinical studies need to aver its impact.


1. Murakami S and Miki Y. Human skin histology using high-resolution echography.J Clin Ultrasound 1989; 2: 77-82.
2. Tran K T, Griffith L, Wells A. Extracellular matrix signaling through growth factor receptors during wound healing.Wound Repair Regen 2004; 3: 262-268.
3. Wang Z, Qian Y, Li L, Pan L, Njunge LW, Dong L, et al. Evaluation of emulsion electrospun polycaprolactone/ hyaluronan/epidermal growth factor nanofibrous scaffolds for wound healing.J Biomater Appl 2016; 6: 686-698.
4. Tadokoro T, Kobayashi N, Zmudzka BZ, Ito S, Wakamatsu K, Yamaguchi Y, et al. UV-induced DNA damage and melanin content in human skin differing in racial/ethnic origin. Faseb j 2003; 9: 1177-1179.
5. Ando H, Kondoh H, Ichihashi M, Hearing VJ. Approaches to identify inhibitors of melanin biosynthesis via the quality control of tyrosinase.J Invest Dermatol 2007; 4: 751-761.
6. Chang TS. An updated review of tyrosinase inhibitors.Int J Mol Sci 2009; 6: 2440-75.
7. Baek S H, Nam I J, Kwak H S, Kim K C, and Lee S H. Cellular anti-melanogenic effects of a Euryale ferox seed extract ethyl acetate fraction via the lysosomal degradation machinery.Int J Mol Sci 2015; 5: 9217-35.
8. Wu ZS, Zhang M, Wang SJ. Effects of high-pressure argon and nitrogen treatments on respiration, browning and antioxidant potential of minimally processed pineapples during shelf life.J Sci Food Agric 2012; 11: 2250-9.
9. Feyzabadi Z, Jafari F, Kamali SH, Ashayeri H, Badiee Aval S, Esfahani MM, Efficacy of Viola odorata in treatment of chronic insomnia.Iran Red Crescent Med J 2014; 12: e17511.
10. Zarrabi M, Dalirfardouei R, Sepehrizade Z, and Kermanshahi R K. Comparison of the antimicrobial effects of semipurified cyclotides from Iranian Viola odorata against some of plant and human pathogenic bacteria.J Appl Microbiol 2013; 2: 367-75.
11. Koochek M, Pipelzadeh M, and Mardani H. The effectiveness of Viola odorata in the prevention and treatment of formalin-induced lung damage in the rat.J Herbs Spices Med Plants 2003; 2: 95-103.
12. Lindholm P, Goransson U, Johansson S, Claeson P, Gullbo J, Larsson R, et al. Cyclotides: a novel type of cytotoxic agents.Mol Cancer Ther 2002; 6: 365-9.
13. Gerlach SL, Rathinakumar R, Chakravarty G, Goransson U, Wimley WC, Darwin S, et al. Anticancer and chemosensitizing abilities of cycloviolacin 02 from Viola odorata and psyle cyclotides from Psychotria leptothyrsa. Biopolymers 2010; 5: 617-25.
14. Siddiqi HS, Mehmood MH, Rehman NU, Gilani AH. Studies on the antihypertensive and antidyslipidemic activities of Viola odorata leaves extract.Lipids Health Dis 2012; 6.
15. Monadi A,Rezaie A. Evaluation of sedative and pre-anesthetic effects of Viola odorata Linn. extract compared with diazepam in rats.Bull Environ Pharmacol Life Sci 2013; 7: 125-131.
16. Ganapaty S, Dash G K, Subburaju T, Suresh P. Diuretic, laxative and toxicity studies of Cocculus hirsutus aerial parts.Fitoterapia 2002; 1: 28-31.
17. Ebrahimzadeh MA, Nabavi SM, Nabavi SF, Bahramian F, Bekhradnia AR. Antioxidant and free radical scavenging activity of H. officinalis L. var. angustifolius, V. odorata, B. hyrcana and C. speciosum.Pak J Pharm Sci 2010; 1: 29-34.
18. Cestari TF, Dantas LP, and Boza JC. Acquired hyperpigmentations.An Bras Dermatol 2014; 1: 11-25.
19. Sadraei H, Asghari G, and Emami S. Effect of Rosa damascena Mill. flower extract on rat ileum.Res Pharm Sci 2013; 4: 277-284.
20. Seidel V. Initial and bulk extraction. In: Sarker SD, Latif Z, Gray AI, editors. Natural Product Isolation. 2nd ed.Humana Press 2006; 27-46.
21. Shafaei-Bajestani N, Emami S A, Asili J, Tayarani-Najaran Z. Anti-apoptotic effect of taxodione on serum/glucose deprivation-induced PC12 cells death.Cell Mol Neurobiol 2014; 8: 1103-9.
22. Qiao Z, Koizumi Y, Zhang M, Natsui M, Flores M J, Gao L, et al. Anti-melanogenesis effect of Glechoma hederacea L. extract on B16 murine melanoma cells.Biosci Biotechnol Biochem 2012; 10: 1877-83.
23. Prakash L and Majeed M. Multifunctional skin tone lighteners from nature: An overview.Eurocosmetics 2009; 19-23.
24. Aranda A, Sequedo L, Tolosa L, Quintas G, Burello E, Castell JV, and Gombau L. Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay: a quantitative method for oxidative stress assessment of nanoparticle-treated cells.Toxicol in vitro 2013; 2: 954-63.
25. Huang HC, Hsieh WY, Niu YL, Chang TM. Inhibition of melanogenesis and antioxidant properties of Magnolia grandiflora L. flower extract.BMC Complement Altern Med 2012; article number 72.
26. Hearing VJ. Determination of melanin synthetic pathways. J Invest Dermatol 2011; E1: E8-e11.
27. Eller MS, Gilchrest BA. Tanning as part of the eukaryotic SOS response.Pigment Cell Res 2000; 94-7.
28. Gilchrest BA, Park HY, Eller MS, Yaar M. Mechanisms of ultraviolet light-induced pigmentation.Photochem Photobiol 1996; 1: 1-10.
29. Coelho SG, Zhou Y, Bushar HF, Miller SA, Zmudzka BZ, Hearing VJ, et al. Long-lasting pigmentation of human skin, a new look at an overlooked response to UV.Pigment Cell Melanoma Res 2009; 2: 238-41.
30. Solano F, Briganti S, Picardo M, Ghanem G. Hypopigmenting agents: an updated review on biological, chemical and clinical aspects.Pigment Cell Res 2006; 6: 550-71.
31. Kahn V. Effect of kojic acid on the oxidation of DL-DOPA, norepinephrine, and dopamine by mushroom tyrosinase.Pigment Cell Res 1995; 5: 234-40.
32. Cabanes J, Chazarra S, Garcia-Carmona F. Kojic acid, a cosmetic skin whitening agent, is a slow-binding inhibitor of catecholase activity of tyrosinase.J Pharm Pharmacol 1994; 12: 982-5.
33. Nakagawa M, Kawai K, Kawai K. Contact allergy to kojic acid in skin care products.Contact Dermatitis 1995; 1: 9-13.
34. Lawrence N, Bligard CA, Reed R, Perret WJ. Exogenous ochronosis in the United States.J Am Acad Dermatol 1988; 2: 1207-11.
35. Nordlund JJ, Grimes PE, and Ortonne JP. The safety of hydroquinone.J Eur Acad Dermatol Venereol 2006; 7: 781-7.
36. Kamakshi R. Fairness via formulations: a review of cosmetic skin-lightening ingredients.J Cosmet Sci 2012; 1: 43-54.
37. Naish-Byfield S, Cooksey C, Latter A, Johnson C,Riley P. In vitro assessment of the structure-activity relationship of tyrosinase-dependent cytotoxicity of a series of substituted phenols.Melan res 1991; 4: 273-288.
38. Halaban R, Patton RS, Cheng E, Svedine S, Trombetta
ES, Wahl ML, et al. Abnormal acidification of melanoma cells induces tyrosinase retention in the early secretory pathway. J Biol Chem 2002; 17: 14821-8.
39. Pompella A, Visvikis A, Paolicchi A, De Tata V, Casini AF. The changing faces of glutathione, a cellular protagonist.Biochem Pharmacol 2003; 8: 1499-1503.
40. Draelos ZD. Novel topical therapies in cosmetic dermatology.Curr Prob Dermatol 2000; 5: 235-239.
41. Katsambas A D and Stratigos A J. Depigmenting and bleaching agents: coping with hyperpigmentation.Clin Dermatol 2001; 4: 483-488.
42. Mason HS. The chemistry of melanin; mechanism of the oxidation of dihydroxyphenylalanine by tyrosinase.J Biol Chem 1948; 1: 83-99.