Archives of Dermatological Research

, Volume 302, Issue 3, pp 191–200 | Cite as

Rhodiola rosea ability to enrich cellular antioxidant defences of cultured human keratinocytes

  • Cinzia Calcabrini
  • Roberta De Bellis
  • Umberto Mancini
  • Luigi Cucchiarini
  • Lucia Potenza
  • Roberta De Sanctis
  • Vania Patrone
  • Carla Scesa
  • Marina Dachà
Original Paper
First Online:
Received:
Revised:
Accepted:

Abstract

Keratinocytes are cells strongly exposed to oxidative stress, but normally good equipped for antioxidant responses. However, it has long been suggested that exogenous antioxidants could play a useful role in minimizing the adverse skin responses associated with such oxidant species. In this work it was paid attention to the extract of Rhodiola rosea L. roots by using the phytocomplex as a whole because of the important activity of its composition and mutual distribution of its components. We have measured the protection afforded by the extract to reduced glutathione levels, glyceraldehyde-3-phosphate dehydrogenase activity, and thiobarbituric acid reactive substances levels in cultured human keratinocytes (NCTC 2544) exposed to different oxidative insults: Fe(II)/ascorbate, Fe(II)/H2O2, and tert-butyl-hydroperoxide. We also have investigated the influence of the R. rosea extract on the production of intracellular reactive oxygen species and on the activity of antioxidant enzymes (catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase). Furthermore, we have demonstrated that R. rosea extract was able to increase in a time- and dose-dependent manner the activity of the trans plasma membrane oxido reductase activity as an indirect evaluation of the intracellular redox status and this effect was already evident with small concentration of the extract and in a long time. As a result, NCTC 2544 are able to better counteract to several oxidative insults if incubated with R. rosea extract demonstrating a very good antioxidant activity of this phytocomplex.

Keywords

Antioxidant enzymes Human keratinocytes Oxidative stress Plasma membrane oxidoreductase Reduced glutathione Rhodiola rosea L. 
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Notes

Acknowledgment

We thank Specchiasol S.r.L. (Verona, Italy) for kindly providing Rhodiola rosea roots.

References

  1. 1.
    Avron M, Shavit N (1963) A sensitive simple method for determination of ferricyanide. Anal Biochem 6:549–555CrossRefPubMedGoogle Scholar
  2. 2.
    Babich H, Visioli F (2003) In vitro cytotoxicity to human cells in culture of some phenolics from olive oil. Farmaco 58(5):403–407CrossRefPubMedGoogle Scholar
  3. 3.
    Battistelli M, De Sanctis R, De Bellis R, Cucchiarini L, Dachà M, Gobbi P (2005) Rhodiola rosea as antioxidant in red blood cells: ultrastructural and hemolytic behaviour. Eur J Histochem 49(3):243–254PubMedGoogle Scholar
  4. 4.
    Baty JW, Hampton MB, Winterbourn CC (2005) Proteomic detection of hydrogen peroxide-sensitive thiol proteins in Jurkat cells. Biochem J 389:785–795CrossRefPubMedGoogle Scholar
  5. 5.
    Beutler E (1975) Red cell metabolism. A manual of biochemical methods. Grune and Strattom, New YorkGoogle Scholar
  6. 6.
    Borenfreund E, Puerner JA (1985) Toxicity determined in vitro by morphological alterations and neutral red absorption. Toxicol Lett 24(2–3):119–124CrossRefPubMedGoogle Scholar
  7. 7.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of proteins utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  8. 8.
    Chen TS, Liou SY, Chang YL (2008) Antioxidant evaluation of three adaptogen extracts. Am J Chin Med 36(6):1209–1217CrossRefPubMedGoogle Scholar
  9. 9.
    De Bellis R, De Sanctis R, Cucchiarini L, Mancini U, Scesa C, Dachà M (2005) Rhodiola rosea as a nutraceutical: from an adaptogen to an excellent antioxidant phytocomplex. In: Pandalai SG (ed) Recent research developments in molecular & cellular biochemistry vol 2, Part II. Research Signpost, Kerala, India, pp 225–236Google Scholar
  10. 10.
    De Sanctis R, De Bellis R, Scesa C, Mancini U, Cucchiarini L, Dachà M (2004) In vitro protective effect of Rhodiola rosea extract against hypochlorous acid-induced oxidative damage in human erythrocytes. BioFactors 20:147–159CrossRefPubMedGoogle Scholar
  11. 11.
    Djavaheri-Mergny M, Mergny JL, Bertrand F, Santus R, Mazière C, Dubertret L, Mazière JC (1996) Ultraviolet-A induces activation of AP-1 in cultured human keratinocytes. FEBS Lett 384:92–96CrossRefPubMedGoogle Scholar
  12. 12.
    Fiorani M, Accorsi A (2005) Dietary flavonoids as intracellular substrates for an erythrocyte trans-plasma membrane oxidoreductase activity. Br J Nutr 94(3):338–345CrossRefPubMedGoogle Scholar
  13. 13.
    Fiorani M, De Sanctis R, De Bellis R, Dachà M (2002) Intracellular flavonoids as electron donors for extracellular ferricyanide reduction in human erythrocytes. Free Radic Biol Med 32(1):64–72CrossRefPubMedGoogle Scholar
  14. 14.
    Fiorani M, De Sanctis R, Scarlatti F, Stocchi V (1998) Substrates of hexokinase, glucose-6-phosphate dehydrogenase, and glyceraldehyde-3-phosphate dehydrogenase prevent the inhibitory response induced by ascorbic acid/iron and dehydroascorbic acid in rabbit erythrocytes. Arch Biochem Biophys 356(2):159–166CrossRefPubMedGoogle Scholar
  15. 15.
    Ganzera M, Yayla Y, Khan IA (2001) Analysis of the marker compounds of Rhodiola rosea L. (golden root) by reversed phase high performance liquid chromatography. Chem Pharm Bull (Tokyo) 49(4):465–467CrossRefGoogle Scholar
  16. 16.
    Gaullier JM, Valla A, Bazin M, Giraud M, Dubertret L, Santus R (1997) N-conjugates of 2,5-disubstituted pyrrole and glutathione evaluation of their potency as antioxidants against photosensitization of NCTC 2544 keratinocytes by excess endogenous protoporphyrin IX. J Photochem Photobiol B 39:24–29CrossRefPubMedGoogle Scholar
  17. 17.
    He YY, Huang JL, Ramirez DC, Chignell CF (2003) Role of reduced glutathione efflux in apoptosis of immortalized human keratinocytes induced by UVA. J Biol Chem 278(10):8058–8064CrossRefPubMedGoogle Scholar
  18. 18.
    Hyun DH, Hernandez JO, Mattson MP, de Cabo R (2006) The plasma membrane redox system in aging. Ageing Res Rev 5:209–220CrossRefPubMedGoogle Scholar
  19. 19.
    Iguchi H, Kojo S, Ikeda M (1993) Lipid peroxidation and disintegration of the cell membrane structure in cultures of rat lung fibroblasts treated with asbestos. J Appl Toxicol 13:269–275CrossRefPubMedGoogle Scholar
  20. 20.
    Kaul N, Choi J, Forman HJ (1998) Transmembrane redox signaling activates NF-kappaB in macrophages. Free Radic Biol Med 24(1):202–207CrossRefPubMedGoogle Scholar
  21. 21.
    Kelly GS (2001) Rhodiola rosea: a possible plant adaptogen. Altern Med Rev 6(3):293–302PubMedGoogle Scholar
  22. 22.
    Kucinskaite A, Briedis V, Savickas A (2004) Experimental analysis of therapeutic properties of Rhodiola rosea L. and its possible application in medicine. Medicina (Kaunas) 40(7):614–619Google Scholar
  23. 23.
    Li W, Dou D, Koike K (2008) Revised absolute stereochemistry of rhodiolosides A-D, rhodiolol A and sachalinol A from Rhodiola rosea. Chem Pharm Bull 56(7):1047–1048CrossRefPubMedGoogle Scholar
  24. 24.
    Linh PT, Kim YH, Hong SP (2000) Quantitative determination of salidroside and tyrosol from the underground part of Rhodiola rosea by high performance liquid chromatography. Arch Pharm Res 23:349–352CrossRefPubMedGoogle Scholar
  25. 25.
    Mao Y, Li Y, Yao N (2007) Simultaneous determination of salidroside and tyrosol in extracts of Rhodiola L. by microwave assisted extraction and high-performance liquid chromatography. J Pharm Biomed Anal 45(3):510–515CrossRefPubMedGoogle Scholar
  26. 26.
    May JM, Qu ZC, Cobb CE (2004) Human erythrocyte recycling of ascorbic acid: relative contributions from the ascorbate free radical and dehydroascorbic acid. J Biol Chem 279(15):14975–14982CrossRefPubMedGoogle Scholar
  27. 27.
    May JM (1999) Is ascorbic acid an antioxidant for the plasma membrane? Faseb J 13:995–1006PubMedGoogle Scholar
  28. 28.
    Mazzola JL, Sirover MA (2003) Subcellular alteration of glyceraldehyde-3-phosphate dehydrogenase in Alzheimer’s disease fibroblasts. J Neurosci Res 71(2):279–285CrossRefPubMedGoogle Scholar
  29. 29.
    Merker MP, Bongard RD, Kettenhofen NJ, Okamoto Y, Dawson CA (2002) Intracellular redox status affects transplasma membrane electron transport in pulmonary arterial endothelial cells. Am J Physiol Lung Cell Mol Physiol 282(1):L36–L43PubMedGoogle Scholar
  30. 30.
    Morini F, Dusatti F, Bonina FP, Saija A, Ferro M (2000) Iron-induced lipid peroxidation in human skin-derived cell lines: protection by a red orange extract. Altern Lab Anim 28(3):427–433Google Scholar
  31. 31.
    Mosmann T (1983) Rapid colorimetric assay for cellular growth and servival: application to proliferation and cytotoxicity assays. J Immunol Methods 65(1–2):55–63CrossRefPubMedGoogle Scholar
  32. 32.
    Nardini M, Pisu P, Gentili V, Natella F, Di Felice M, Piccolella E, Scaccini C (1998) Effect of caffeic acid on tert-butyl hydroperoxide-induced oxidative stress in U937. Free Radic Biol Med 25(9):1098–1105CrossRefPubMedGoogle Scholar
  33. 33.
    Navas P, Estévez A, Buròn MI, Villalba JM, Crane FL (1988) Cell surface glycoconjugates control the activity of the NADH-ascorbate free radical reductase of rat liver plasma membrane. Biochem Biophys Res Commun 154(3):1029–1033CrossRefPubMedGoogle Scholar
  34. 34.
    Paoletti F, Aldinucci D, Mocali A, Caparrini A (1986) A sensitive spectrophotometric method for the determination of superoxide dismutase activity in tissue extracts. Anal Biochem 154:536–541CrossRefPubMedGoogle Scholar
  35. 35.
    Roques SC, Landrault N, Teissedre PL, Laurent C, Besancon P, Rouane JM, Caporiccio B (2002) Hydrogen peroxide generation in caco-2 cell culture medium by addition of phenolic compounds: effect of ascorbic acid. Free Radic Res 36(5):593–599CrossRefPubMedGoogle Scholar
  36. 36.
    Sestili P, Brandi G, Brambilla L, Cattabeni F, Cantoni O (1996) Hydrogen peroxide mediates the killing of U937 tumor cells elicited by pharmacologically attainable concentrations of ascorbic acid: cell death prevention by extracellular catalase or catalase from cocultured erythrocytes or fibroblasts. J Pharm Exp Ther 277(3):1719–1725Google Scholar
  37. 37.
    Schriner SE, Avanesian A, Liu Y, Luesch H, Jafari M (2009) Protection of human cultured cells against oxidative stress by Rhodiola rosea without activation of antioxidant defences. Free Radic Biol Med 47(5):577–584CrossRefPubMedGoogle Scholar
  38. 38.
    Skarpanska-Stejnborn A, Pilaczynska-Szczesniak L, Basta P, Deskur-Smielecka E (2009) The influence of supplementation with Rhodiola rosea L. Extract on selected redox parameters in professional rowers. Int J Sport Nutr Ex Metab 19:186–199Google Scholar
  39. 39.
    Stearns DM, Kennedy LJ, Courtney KD, Giangrande PH, Phieffer LS, Wetterhahn KE (1995) Reduction of chromium(VI) by ascorbate leads to chromium-DNA binding and DNA strand breaks in vitro. Biochemistry 34(3):910–919CrossRefPubMedGoogle Scholar
  40. 40.
    Tobi SE, Paul N, McMillan TJ (2000) Glutathione modulates the level of free radicals produced in UVA-irradiated cells. J Photochem Photobiol B 57:102–112CrossRefPubMedGoogle Scholar
  41. 41.
    Vessey DA, Lee KH, Blacker KL (1992) Characterization of the oxidative stress initiated in cultured human keratinocytes by treatment with peroxides. J Invest Dermatol 99:859–863CrossRefPubMedGoogle Scholar
  42. 42.
    Vissers MC, Carr AC, Chapman AL (1998) Comparison of human red cell lysis by hypochlorous and hypobromous acids: insights into the mechanism of lysis. Biochem J 330:131–138PubMedGoogle Scholar
  43. 43.
    Wang H, Joseph JA (1999) Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader. Free Radic Biol Med 27:612–616CrossRefPubMedGoogle Scholar
  44. 44.
    Wiegant FA, Surinova S, Ytsma E, Limandjaja G, Langelaar-Makkinje M, Wikman G, Post JA (2009) Plant adaptogens increase lifespan and stress resistance in C. elegans. Biogerontol 10:27–42CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Cinzia Calcabrini
    • 1
  • Roberta De Bellis
    • 1
  • Umberto Mancini
    • 1
  • Luigi Cucchiarini
    • 1
  • Lucia Potenza
    • 1
  • Roberta De Sanctis
    • 1
  • Vania Patrone
    • 2
  • Carla Scesa
    • 3
  • Marina Dachà
    • 4
  1. 1.Istituto di Chimica Biologica “Giorgio Fornaini”Università degli Studi di Urbino “Carlo Bo”UrbinoItaly
  2. 2.Istituto di IgieneUniversità degli Studi di Urbino “Carlo Bo”UrbinoItaly
  3. 3.Facoltà di Medicina e ChirurgiaUniversità Cattolica del Sacro CuoreRomeItaly
  4. 4.Università Campus Bio-MedicoRomeItaly

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