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Journal of Wood Science

Official Journal of the Japan Wood Research Society

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Effect of Pinus radiate bark extracts with different molecular weight distributions on cell growth of NIH/3T3 fibroblasts and dendrite retraction of B16 melanoma cells

Journal of Wood Science volume 57, pages 415–420 (2011)Cite this article

Abstract

Hot water extract (HWE) of Pinus radiata bark was separated into monomeric polyphenol (MPP), oligomeric proanthocyanidin (OPA), and polymeric proanthocyanidin (PPA) fractions by monitored chromatography using a Sephadex LH 20 column and an UV detector at 250 nm. The effects of these fractions on NIH/3T3 fibroblasts and B16 melanoma cells were examined by evaluating cell viability, melanogenesis (melanin content), morphological changes, and tyrosinase inhibitory activity. The polyphenolic fractions had a proliferation effect on fibroblasts, with cell growth increasing significantly (P < 0.01) even at the high concentration of 1250 μg/ml. At 125 μg/ml, HWE, MPP, and OPA had no effect on melanogenesis, whereas PPA significantly upregulated melanogenesis (P < 0.05). Melanogenesis was significantly upregulated in melanoma cells treated with these fractions at the high concentration of 600 μg/ml (P < 0.01). B16 melanoma cells cultured with the proanthocyanidin (PA)-rich fractions (HWE, OPA, and PPA) showed marked dendrite retraction, leading to significant morphological transformation. OPA in particular showed colored adhesion on the surface of melanoma cells. All four fractions significantly inhibited mushroom tyrosinase activity when compared to arbutin and ascorbic acid 2-glucoside (P < 0.05). The tyrosinase inhibitory activity of OPA and PPA did not differ significantly (P = 0.11), indicating that the inhibitory effects had a low correlation with molecular weight distribution. The inhibition kinetics of PPA determined using a Lineweaver-Burk plot indicated that PPA is a noncompetitive inhibitor of l-3,4-dihydroxyphenylalanine oxidation by mushroom tyrosinase.

References

  1. Kofujita H, Ettyu K, Ota M (1999) Characterization of the major components in bark from five Japanese tree species for chemical utilization. Wood Sci Technol 33:223–228

    Article  CAS  Google Scholar 

  2. Hon DNS, Shiraishi N (1991) Wood and cellulosic chemistry. Marcel Dekker, New York

    Google Scholar 

  3. Saliou C, Rimbach G, Moini H, McLaughlin L, Hosseini S, Lee J, Watson RR, Packer L (2001) Solar ultraviolet-induced erythema in human skin and nuclear factor-kappa-B-dependent gene expression in keratinocytes are modulated by a French maritima pine bark extract. Free Rad Biol Med 30:154–160

    Article  CAS  PubMed  Google Scholar 

  4. Svobodová A, Psotová J, Walterová D (2003) Natural phenolics in the prevention of UV-induced skin damage. Biomed Papers 147:137–145

    Article  Google Scholar 

  5. Yamakoshi J, Otsuka F, Sano A, Tokutake S, Saito M, Kikuchi M, Kubota Y (2003) Lightening effect on ultraviolet-induced pigmentation of guinea pig skin by oral administration of a proanthocyanidinrich extract from grape seeds. Pigment Cell Res 16:629–638

    Article  CAS  PubMed  Google Scholar 

  6. Ku CS, Mun SP (2007) Characterization of proanthocyanidin in hot water extract isolated from Pinus radiata bark. Wood Sci Technol 41:235–247

    Article  CAS  Google Scholar 

  7. Ku CS, Mun SP (2008) Antioxidant properties of monomeric, oligomeric, and polymeric fractions in hot water extract from Pinus radiata bark. Wood Sci Technol 42:47–60

    Article  CAS  Google Scholar 

  8. Ohara S, Suzuki K, Ohira T (1994) Condensed tannins from Acacia mearnsii and their biological activities. Mokuzai Gakkaishi 40:1363–1374

    CAS  Google Scholar 

  9. Kraus TEC, Yu Z, Preston R, Dahlgren RA, Zasoski RJ (2003) Linking chemical reactivity and protein precipitation to structural characteristics of foliar tannins. J Chem Ecol 29:703–730

    Article  CAS  PubMed  Google Scholar 

  10. Shoji T, Masumoto S, Moriichi N, Kobori M, Kanda T, Shinmoto H, Tsushida T (2005) Procyanidin trimers to pentamers fractionated from apple inhibit melanogenesis in B16 mouse melanoma cells. J Agric Food Chem 53:6105–6111

    Article  CAS  PubMed  Google Scholar 

  11. Perluigi M, De Marco F, Foppoli C, Coccia R, Blarzino C, Marcante ML, Cini C (2003) Tyrosinase protects human melanocytes from ROS-generating compounds. Biochem Biophys Res Commun 305:250–256

    Article  CAS  PubMed  Google Scholar 

  12. Nagata H, Takekoshi S, Takeyama R, Homma T, Osamura RY (2004) Quercetin enhances melanogenesis by increasing the activity and synthesis of tyrosinase in human melanoma cells and in normal human melanocytes. Pigment Cell Res 17:66–73

    Article  CAS  PubMed  Google Scholar 

  13. Hakozaki T, Minwalla L, Zhuang J, Chhoa M, Matsubara A, Miyamoto K (2002) The effect of niacinamide on reducing cutaneous pigmentation and suppression of melanosome transfer. Br J Dermatol 147:20–31

    Article  CAS  PubMed  Google Scholar 

  14. Ito Y, Kanamaru A, Tada A (2006) Centaureidin promotes dendrite retraction of melanocytes by activating Rho. Biochim Biophys Acta 1760:487–494

    Article  CAS  PubMed  Google Scholar 

  15. Ito Y, Kanamaru A, Tada A (2006) Effects of methylophiopogonanone B on melanosome transfer and dendrite retraction. J Dermatol Sci 42:68–70

    Article  CAS  PubMed  Google Scholar 

  16. Nakamura Y, Torikai K, Ohigashi H (2001) A catechol antioxidant protocatechuic acid potentiates inflammatory leukocyte-derived oxidative stress in mouse skin via a tyrosinase bioactivation pathway. Free Rad Biol Med 30:967–978

    Article  CAS  PubMed  Google Scholar 

  17. Ito S (2003) A chemist’s view of melanogenesis. Pigment Cell Res 16:230–236

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Institute of Agricultural Science and Technology, Chonbuk National University, Jeonju, 561-756, Korea

    Chang Sub Ku

  2. Department of Wood Science and Technology, College of Agriculture and Life Sciences, Chonbuk National University, Jeonju, 561-756, Korea

    Sung Phil Mun

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  1. Chang Sub Ku
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  2. Sung Phil Mun
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Correspondence to Sung Phil Mun.

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Ku, C.S., Mun, S.P. Effect of Pinus radiate bark extracts with different molecular weight distributions on cell growth of NIH/3T3 fibroblasts and dendrite retraction of B16 melanoma cells. J Wood Sci 57, 415–420 (2011). https://doi.org/10.1007/s10086-011-1193-y

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  • DOI: https://doi.org/10.1007/s10086-011-1193-y

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