Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Mechanisms of Suppression of Matrix Metalloproteinases in UVB-Irradiated HaCaT Keratinocytes of Colored Rice Varieties

UVB에 조사된 HaCaT Keratinocytes에서의 유색미에 의한 Matrix Metalloproteinases 발현억제 메커니즘

  • Choi, Eun-Young (Department of Cosmeceutical Science, Daegu Haany University) ;
  • Lee, Jae-Bong (Department of Cosmeceutical Science, Daegu Haany University) ;
  • Kim, Do-Hoon (Department of Genetics Engineering, College of Natural Resource & Life Science, Dong-A University) ;
  • Kwon, Yong-Sham (Department of Genetics Engineering, College of Natural Resource & Life Science, Dong-A University) ;
  • Cheon, Jung-Yoon (Quality Assurance Team, Thuringen Korea Co., Ltd.) ;
  • Lee, Jin-Tae (Department of Cosmeceutical Science, Daegu Haany University)
  • 최은영 (대구한의대학교 화장품약리학과) ;
  • 이재봉 (대구한의대학교 화장품약리학과) ;
  • 김도훈 (동아대학교 생명자원과학대학 유전공학과) ;
  • 권용삼 (동아대학교 생명자원과학대학 유전공학과) ;
  • 천정윤 ((주)튜링겐코리아 품질보증팀) ;
  • 이진태 (대구한의대학교 화장품약리학과)
  • Received : 2017.02.09
  • Accepted : 2017.04.10
  • Published : 2017.05.31
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Abstract

In this study, we investigated the anti-oxidant activities [electron-donating ability (EDA), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging ability, and reactive oxygen species (ROS) inhibitory activity], anti-wrinkle activities [collagenase inhibitory activity, suppression and/or phosphorylation of matrix metalloproteinases (MMPs), and mitogen-activated protein (MAP) kinase activity], and mRNA expression levels using reverse transcription-polymerase chain reaction (RT-PCR) assay in ultraviolet (UV) B ray ($50mJ/cm^2$)-irradiated human keratinocyte HaCaT cells. Josaengheugchal, Sinneungheugchal (SE), Shintoheug rice, Heugjinju rice, and Heugseol (HE) among colored rice varieties were reported to have excellent antioxidant properties. In the EDA and ABTS radical scavenging assays, extracts of the five colored rice varieties had scavenging activities of 72% at concentrations higher $50{\mu}g/mL$. In the collagenase inhibition assay, ethanol extracts of the five colored rice varieties showed high inhibitory effects of about 60% at concentrations higher $25{\mu}g/mL$. In the ROS inhibition assay, ethanol extracts of HE and SE showed very excellent inhibition efficacies at all concentrations. We determined molecular biological mechanisms of MMPs (MMP-1, -3, -8, and -13) and mitogen-activated protein kinase (MAPK) with HE, and the results show that HE suppressed expression of MMPs and phosphorylation of MAPK and increased expression of pro-collagen type I in UVB-irradiated cells. It was also confirmed by RT-PCR that HE reduced expression of MMPs mRNA. Therefore, these results suggest that HE has anti-wrinkle and collagen production effects and may be used as a material in the development of functional food and cosmetic industries.

본 연구에서는 선별된 유색미 5품종에 대한 자외선 조사에 따른 노화 억제 기전을 확인하고자 항산화 활성과 활성산소종(ROS) assay를 분석하였으며, 주름형성을 유발하는 효소인 MMPs와 그 upstream인 MAPK pathway(ERK, JNK 및 p38)에서의 인산화 발현을 얼마나 저해하는지를 분석하였다. 유색미 5품종 가운데 전자공여능과 ABTS 소거활성 결과, 신토흑미 에탄올 추출물(SRE)과 흑설 에탄올 추출물(HE), SRE와 흑설 열수 추출물(HW)이 각각 뛰어난 소거활성을 보였으며, collagenase 저해 활성 결과 HE가 전 농도에서 고르게 높은 저해효과를 나타내었다. 또한, ROS assay에서 SE와 HE가 전 농도에서 매우 효과적인 ROS 생성억제를 나타냄에 따라 우리는 선별된 유색미 가운데 HE를 이용하여 MMPs와 MAPK의 발현억제 pathway를 확인하였다. 그 결과 pro-collagen type I은 $100{\mu}g/mL$의 농도에서 발현이 증가하였고, MMP-1과 -13은 농도 의존적으로 발현이 억제되었으며, ERK와 p38의 인산화 또한 농도 의존적으로 억제하고 있음을 확인하였다. 이러한 결과에 따라 MMPs의 mRNA 발현을 확인하기 위한 RT-PCR을 실시하였으며, MMP-1과 3의 mRNA의 발현은 농도 의존적으로 감소하였고 MMP-9는 HE를 $100{\mu}g/mL$의 농도로 처치한 경우에 발현이 감소하였음을 확인하였다. 선별된 유색미 5종의 에탄올 추출물은 항산화 활성이 우수하였고, 특히 HE의 주름억제 활성 효능을 확인함에 따라 기능성 식품 화장품 분야에서 주름개선 및 항노화 소재로서의 가능성을 확인하였다.

Keywords

References

  1. Her YM, Lee SY, Lee SY. 2007. Preparation of functional nutrient enriched rices by coating method. Food Eng Prog 11: 185-197.
  2. Ryu SN, Park SZ, Kang SS. 2005. Studies on exploration and expansive use of genetic variation of functional substances in rice. Rural Development Administration, Jeonju, Korea.
  3. Nam SH, Chang SM, Kang MY. 2003. Varietal difference in antioxidative activity of ethanolic extracts from colored rice bran. J Korean Soc Agric Chem Biotechnol 46: 16-22.
  4. Kang MY, Nam YJ, Nam SH. 2005. Screening of antioxidation-related functional components in brans of the pigmented rices. J Korean Soc Appl Biol Chem 48: 233-239.
  5. Choi HC, Oh SK. 1996. Diversity and function of pigments in colored rice. Korean J Corp Sci 41: 1-9.
  6. Ling WH, Wang LL, Ma J. 2002. Supplementation of the black rice outer layer fraction to rabbits decrease atherosclerotic plaque formation and increases antioxidant status. J Nutr 132: 20-26. https://doi.org/10.1093/jn/132.1.20
  7. Slavin JL, Martini MC, Jacobs Jr DR, Marquart L. 1999. Plausible mechanisms for the protectiveness of whole grains. Am J Clin Nutr 70: 459S-463S. https://doi.org/10.1093/ajcn/70.3.459s
  8. Ryu SN, Han SJ, Park SZ, Kim HR. 2006. Antioxidant activity of blackish purple rice. Korean J Crop Sci 51: 173-178.
  9. Choi SW, Nam SH, Choi HC. 1996. Antioxidative activity of ethanolic extracts of rice brans. Food Sci Biotechnol 5: 305-309.
  10. Tsuda T, Horio F, Osawa T. 1998. Dietary cyanidine-3-O-${\beta}$-D-glucoside increases ex vivo oxidation resistance of serum in rats. Lipids 33: 583-588. https://doi.org/10.1007/s11745-998-0243-5
  11. Nam SH, Kang MY. 1998. Comparison of inhibitory effect of rice bran-extracts of the colored rice cultivars on carcinogenesis. Appl Biol Chem 41: 78-83.
  12. Song SJ, Lee YS, Rhee CO. 2000. Volatile flavor components in cooked black rice. Korean J Food Sci Technol 32: 1015-1021.
  13. Chae S, Piao MJ, Kang KA, Zhang R, Kim KC, Youn UJ, Nam KW, Lee JH, Hyun JW. 2011. Inhibition of matrix metalloproteinase-1 induced by oxidative stress in human keratinocytes by mangiferin isolated from Anemarrhena asphodeloides. Biosci Biotechnol Biochem 75: 2321-2325. https://doi.org/10.1271/bbb.110465
  14. Fisher GJ, Kang S, Varani J, Bata-Csorgo Z, Wan Y, Datta S, Voorhees JJ. 2002. Mechanisms of photoaging and chronological skin aging. Arch Dermatol 138: 1462-1470.
  15. Matrisian LM. 1992. The matrix-degrading metalloproteinases. Bioessays 14: 455-463. https://doi.org/10.1002/bies.950140705
  16. Chiang HM, Chen HC, Lin TJ, Shih IC, Wen KC. 2012. Michelia alba extract attenuates UVB-induced expression of matrix metalloproteinases via MAP kinase pathway in human dermal fibroblasts. Food Chem Toxicol 50: 4260-4269. https://doi.org/10.1016/j.fct.2012.08.018
  17. Fisher GJ, Datta SC, Talwar HS, Wang ZQ, Varani J, Kang S, Voorhees JJ. 1996. Molecular basis of sun-induced premature skin ageing and retinoid antagonism. Nature 379: 335-339. https://doi.org/10.1038/379335a0
  18. Mathews-Roth MM, Michel JL, Wise RJ. 1995. Amelioration of the metabolic defect in erythropoietic protoporphyria by expression of human ferrochelatase in cultured cells. J Invest Dermatol 104: 497-499. https://doi.org/10.1111/1523-1747.ep12605930
  19. Coussens LM, Tinkle CL, Hanahan D, Werb Z. 2000. MMP-9 supplied by bone marrow-derived cells contributes to skin carcinogenesis. Cell 103: 481-490. https://doi.org/10.1016/S0092-8674(00)00139-2
  20. Varani J, Warner RL, Gharaee-Kermani M, Phan SH, Kang S, Chung JH, Wang ZQ, Datta SC, Fisher GJ, Voorhees JJ. 2000. Vitamin A antagonizes decreased cell growth and elevated collagen-degrading matrix metalloproteinases and stimulates collagen accumulation in naturally aged human skin. J Invest Dermatol 114: 480-486. https://doi.org/10.1046/j.1523-1747.2000.00902.x
  21. Zurick KM, Qin C, Bernards MT. 2012. Adhesion of MC3T3-E1 cells bound to dentin phosphoprotein specifically bound to collagen type I. J Biomed Mater Res A 100: 2492-2498.
  22. Park JY, Ham H, Han SI, Oh SH, Song YC, Cho JH, Hur Y, Lee YY, Lee BW, Choi YH. 2016. Comparison of antioxidant components and antioxidant activities of colored rice varieties (Oryza sativa L.) cultivated in Southern plain. J Korean Soc Food Sci Nutr 45: 1214-1220. https://doi.org/10.3746/jkfn.2016.45.8.1214
  23. Blois MS. 1958. Antioxidant determinations by the use of a stable free radical. Nature 181: 1199-1200. https://doi.org/10.1038/1811199a0
  24. Park EJ, Kang MH. 2002. Application of the alkaline comet assay for detecting oxidative DNA damage in human biomonitoring. Korean J Nutr 35: 213-222.
  25. Cannell RJP, Kellam SJ, Owsianka AM, Walker JM. 1988. Results of a large scale screen of microalgae for the production of protease inhibitors. Planta Med 54: 10-14. https://doi.org/10.1055/s-2006-962319
  26. Mosmann T. 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55-63. https://doi.org/10.1016/0022-1759(83)90303-4
  27. Heo JC, Park JY, An SM, Lee JM, Yun CY, Shin HM, Kwon TK, Lee SH. 2006. Anti-oxidant and anti-tumor activities of crude extracts by Gastrodia elata Blume. Korean J Food Preserv 13: 83-87.
  28. Ancerewicz, J, Migliavacca E, Carrupt PA, Testa B, Bree F, Zini R, Tillement JP, Labidalle S, Guyot D, Chauvet-Monges AM, Crevat A, Ridant AL. 1998. Structure-property relationships of trimetazidine derivatives and model compounds as potential antioxidants. Free Radical Biol Med 25: 113-120. https://doi.org/10.1016/S0891-5849(98)00072-0
  29. Song ES, Park SJ, Woo NRA, Won MH, Choi JS, Kim JG, Kang MH. 2005. Antioxidant capacity of colored barley extracts by varieties. J Korean Soc Food Sci Nutr 34: 1491-1497. https://doi.org/10.3746/jkfn.2005.34.10.1491
  30. Woo KS, Jeong EG, Suh SJ, Yang CI, Jeong HS, Kim KJ. 2008. Antioxidant components and antioxidant activities of 70% ethanol extracts on Suweon-511 and Ilpum rice. J Korean Soc Food Sci Nutr 37: 1223-1230. https://doi.org/10.3746/jkfn.2008.37.10.1223
  31. Lee YR, Woo KS, Kim KJ, Son JR, Jeong HS. 2007. Antioxidant activities of ethanol extracts from germinated specialty rough rice. Food Sci Biotechnol 16: 765-770.
  32. Seo WD, Kim JY, Son YC, Cho JH, Jang KC, Han SI, Ra JE, Oh SH, Kang HJ, Kim BJ, Baek NI, Jeong RH, Nam MH. 2013. Comparative analysis of physicochemicals and antioxidative properties in new red rice (Oryza sativa L. cv. Gunganghongmi). J Crop Sci Biotechnol 16: 63-68. https://doi.org/10.1007/s12892-012-0057-3
  33. Fanger GR, Johnson NL, Johnson GL. 1997. MEK kinases are regulated by EGF and selectively interact with Rac/Cdc42. EMBO J 16: 4961-4972. https://doi.org/10.1093/emboj/16.16.4961
  34. Min EG, Kim YH, Kum SI, Han YH. 2004. Inhibition of growth and collagenase activity of the extract from Salvia miltiorrhiza against microorganisms causing periodontal diseases. Korean J Microbiol 40: 111-114.
  35. Kim JH, Na Y, Sim GS, Lee BC, Pyo HB. 2006. Antioxidative and anti-inflammatory effects of Petasites japonicus. J Soc Cosmet Sci Korea 32: 263-267.
  36. Gross J, Lapiere CM. 1962. Collagenolytic activity in amphibian tissues: a tissue culture assay. Proc Natl Acad Sci USA 48: 1014-1022. https://doi.org/10.1073/pnas.48.6.1014
  37. Fisher GJ, Wang ZQ, Datta SC, Varani J, Kang S, Voorhees JJ. 1997. Pathophysiology of premature skin aging induced by ultraviolet light. N Engl J Med 337: 1419-1429. https://doi.org/10.1056/NEJM199711133372003
  38. Brinckerhoff CE, Rutter JL, Benbow U. 2000. Interstitial collagenases as markers of tumor progression. Clin Cancer Res 6: 4823-4830.
  39. Lee SM, Lee SJ, Kwon YY, Baek SH, Kim JS, Sohn HY, Shin WC. 2014. Skin whitening and anti-wrinkle effects of extract from Jubak of oriental herbal liquor. J Korean Soc Food Sci Nutr 43: 1695-1700. https://doi.org/10.3746/jkfn.2014.43.11.1695
  40. Hwang BM, Noh EM, Kim JS, Kim JM, You YO, Hwang JK, Kwon KB, Lee YR. 2013. Curcumin inhibits UVB-induced matrix metalloproteinase-1/3 expression by suppressing the MAPK-p38/JNK pathways in human dermal fibroblasts. Exp Dermatol 22: 371-374. https://doi.org/10.1111/exd.12137
  41. Wenk J, Brenneisen P, Wlaschek M, Poswig A, Briviba K, Oberley TD, Scharffetter-Kochanek K. 1999. Stable overexpression of manganese superoxide dismutase in mitochondria identifies hydrogen peroxide as a major oxidant in the AP-1-mediated induction of matrix-degrading metalloprotease-1. J Biol Chem 274: 25869-25876. https://doi.org/10.1074/jbc.274.36.25869
  42. Brenneisen P, Sies H, Scharffetter-Kochanek K. 2002. Ultraviolet-B irradiation and matrix metalloproteinases: from induction via signaling to initial events. Ann N Y Acad Sci 973: 31-43. https://doi.org/10.1111/j.1749-6632.2002.tb04602.x