Assessment of Eyebright (Euphrasia Officinalis L.) Extract Activity in Relation to Human Corneal Cells Using In Vitro Tests

Abstract

Background: Euphrasia officinalis L. is an herb traditionally used in folk medicine, mainly in the treatment of eye disorders. Aims: The present study analyzed the activity of three extracts of E. officinalis L. (ethanol, ethyl acetate and heptane) on cultured human corneal epithelial cells (10.014 pRSV-T). Study Design: In vitro study. Methods: Toxicity, free radical scavenging activity and the immunomodulatory effects of the extracts were tested using the thiazolyl blue tetrazolium bromide (MTT) or Neutral Red, 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and ELISA tests, respectively. Moreover, nitric oxide levels and cytoskeleton architecture were analyzed after corneal cell incubation with the plant extracts. Results: We show that the biological effect depended on both the concentration and the extraction solvent used. Heptane extracts, distinct from those in ethanol and ethyl acetate, were toxic to 10.014 pRSV-T cells at low concentrations (25 μg/mL) and did not demonstrate free radical scavenging effects. All tested extracts decreased pro-inflammatory cytokine expression (IL-1β, IL-6 and TNF-α) and also anti-inflammatory IL-10 expression by human corneal cells when the extracts were added to the cell culture medium for 24 h. Conclusion: In conclusion, we show that the promising effects of the application of E. officinalis L. preparations as a supplementary therapy for eye disorders are associated with the ethanol and ethyl acetate extracts, not the heptane extract.

Keywords

• Cytotoxicity, Euphrasia officinalis extracts, human corneal cells, immunomodulation, reactive oxygen species scavenging effect

Assessment of Eyebright (Euphrasia Officinalis L.) Extract Activity in Relation to Human Corneal Cells Using In Vitro Tests

Öz

References

1. Petrichenko VM, Sukhinina TV, Babiyan LK, Shramm NI. Chemical com- position and antioxidant properties of biologically active compounds from Euphrasia brevipila. Pharm Chem J 2006;40:312-6. [CrossRef]
2. Stoss M, Michels C, Peter E, Beutke R, Gorter RW. Prospective co- hort trial of Euphrasia single-dose eye drops in conjunctivitis. J Altern Complement Med 2000;6:499-508. [CrossRef]
3. Blazics B, Alberti A, Kéry A. Antioxidant activity of different pheno- lic fractions separated from Euphrasia rostkoviana Hayne. Acta Pharm Hung 2009;79:11-6.
4. Citarasu T. Herbal biomedicines: a new opportunity for aquaculture in- dustry. Aquacult Int 2010;18:403-14. [CrossRef]
5. Kozłowski J, Krajewska A. Ziele świetlika jako leczniczy surowiec roślinny. Farm Pol 1982;38:471-4 (in polish).
6. Porchezhian E, Ansari SH, Shreedharan NKK. Antihyperglycemic activity of Euphrasia officinale leaves. Fitoterapia 2000;71:522-6. [CrossRef]
7. Ríos JL, Recio MC. Medicinal plants and antimicrobial activity. J Eth- nopharmacol 2005;100:80-4. [CrossRef]
8. Trovato A, Monforte MT, Forestieri AM, Pizzimenti F. In vitro anti- mycotic activity of some medicinal plants containing flavonoids. Boll Chim Farm 2000;139:225-7.

9. Dharma P, Singh G. A Review of Plant Species Used to Treat Conjunc- tivitis. Review article. Phytother Res 2002;16:1-22. [CrossRef]
10. Bielory L, Heimall J. Review of complementary and alternative medi- cine in treatment of ocular allergies. Curr Opin Allergy Clin Immunol 2003;5:395-9. [CrossRef]
11. Aburjai T, Natsheh FM. Plants used in cosmetics. Phytother Res 2003;17:987-1000. [CrossRef]
12. Ganbold M, Barker J, Ma R, Jones L, Carew M. Cytotoxicity and bioa- vailability studies on a decoction of Oldenlandia diffusa and its fractions separated by HPLC. J Ethnopharmacol 2010;131:396-403. [CrossRef]
13. Paduch R, Matysik G, Wójciak-Kosior M, Kandefer-Szerszeń M, Skalska-Kamińska A, Nowak-Kryska M, et al. Lamium album extracts express free radical scavenging and cytotoxic activities. Polish J Envi- ron Stud 2008;17:569-80.
14. Revoltella RP, Papini S, Rosellini A, Michelini M. Epithelial stem cells of the eye surface. Cell Prolif 2007;40:445-61. [CrossRef]
15. Wylęgała E, Dobrowolski D, Nowińska A, Tarnawska D. Anterior seg- ment optical coherence tomography in eye injuries. Graefes Arch Clin Exp Ophthalmol 2009;247:451-5. [CrossRef]
16. Chana HS, Schwab L, Foster A. With an eye to good practice. Traditio- nal healers in rural communities. World Health Forum 1994;15:144-6.
17. West AL, Fetters MD, Hemmila MR, Gorenflo DW, Kiyota A, Moroi-Fet- ters S. Herb and vitamin supplementation use among a general ophthalmo- logy practice population. Am J Ophthalmol 2005;139:522-9. [CrossRef]
18. Calixto JB, Campos MM, Otuki MF, Santos ARS. Anti-inflammatory compounds of plant origin. Part II. Modulation of pro-inflammatory cytokines, chemokines and adhesion molecules. Planta Med 2004;70: 93-103. [CrossRef]
19. Shestakova TS, Petrichenko VM, Sukhinina TV. Elemental compositi- on of Euphrasia brevipila herbs and extracts. Pharm Chem J 2008;42: 460-2. [CrossRef]
20. Baudouin C. The pathology of dry eye. Surv Ophthalmol 2001;45 (Suppl. 2):S211-20. [CrossRef]
21. Luo L, Li DQ, Doshi A, Farley W, Corrales RM, Pflugfelder SC. Expe- rimental dry eye stimulates production of inflammatory cytokines and MMP-9 and activates MAPK signaling pathways on the ocular surface. Invest Ophthalmol Vis Sci 2004;45:4293-301. [CrossRef]
22. Yeh S, Song XJ, Farley W, Li DQ, Stern ME, Pflugfelder SC. Apoptosis of ocular surface cells in experimentally induced dry eye. Invest Oph- thalmol Vis Sci 2003;44:124-9. [CrossRef]
23. Achike FI, Kwan Ch-Y. Nitric oxide, human diseases and the herbal pro- ducts that affect the nitric oxide signalling pathway. Clin Exp Pharmacol Physiol 2003;30:605-5. [CrossRef]
24. Kim PKM, Zamora R, Petrosko P, Billiar TR. The regulatory role of nitric oxide in apoptosis. Int Immunopharmacol 2001;1:1421-41. [CrossRef]
25. Schmitt ChA, Dirsch VM. Modulation of endothelial nitric oxide by plant-derived products. Nitric Oxide 2009;21:77-91. [CrossRef]