Effects of Chlorophyll a on UVB-induced Cellular Responses and Type I pN Collagen Synthesis in vitro

클로로필 a가 UVB 유도성 산화적 스트레스와 matrix metalloproteinases (MMPs) 활성화 및 콜라겐 합성에 미치는 영향

  • 전희영 ((주)아모레퍼시픽 기술연구원) ;
  • 김정기 ((주)아모레퍼시픽 기술연구원) ;
  • 서대방 ((주)아모레퍼시픽 기술연구원) ;
  • 이상준 ((주)아모레퍼시픽 기술연구원)
  • Published : 2009.12.31

Abstract

Several studies have demonstrated that chlorophyll has many beneficial properties, including anti-inflammatory, anti-tumor, and antioxidant properties. Chlorophyll has been also shown to have an excellent chemopreventive potential against skin tumor. Its preventive mechanism against skin tumor, however, has not been examined in detail. Furthermore, the effects of chlorophyll on UVB-induced cellular responses have not been investigated. Solar UV radiation, in particular its UVB component, is the primary cause of many adverse biological effects, which is responsible for the photoaging and skin cancer. We investigated the preventive effects of chlorophyll a on UVB-mediated responses in human immortalized HaCaT kerationocytes and normal human fibroblast. We found that treatment of chlorophyll a markedly inhibited UVB-induced generation of reactive oxygen species and lipid peroxidation. Chlorophyll a also prevented UVB-induced MMP-1 expression and MMP-2 activation and increased Type I pN collagen synthesis. These results suggest that chlorophyll a is a potent candidate for the prevention and treatment of UVB-induced skin cancer and photoaging.

본 연구에서는 식물에서 추출된 천연 클로로필 a를 이용하여 UVB에 의해 유도되는 산화적 손상 및 MMPs의 변화를 평가하고, 콜라겐의 합성에 미치는 영향을 알아보고자 하였다. 본 연구의 결과를 종합해 보면, 클로로필 a는 UVB에 의해 유도되는 ROS의 발생을 저해함으로써 그로부터 발생할 수 있는 일련의 물리화학적 혹은 생물학적 신호 전달을 방지하는 효능을 가지며, 지질 과산화 역시 억제하는 효과를 지닌다. 또한 피부의 콜라겐, 탄력 섬유 등 기질 단백질을 분해하는 효소인 MMPs의 발현 또는 활성을 억제하고 콜라겐의 합성을 촉진하는 효과를 보여 피부 광노화에 대한 예방제로서 클로로필의 임상 적용의 가능성을 확인 할 수 있었다. 현재까지 보고된 피부에 대한 클로로필의 효능은 DMBA 또는 TPA 등의 화학 물질로 유발된 피부암을 예방하거나 억제하는 효능에 대한 연구가 대부분이었으며, 본 연구와 같이 UVB에 의해 유발되는 세포 반응에 대한 클로로필 a의 효능연구는 체계적으로 이루어진 바가 없다. 따라서 본 연구는 클로로필의 피부암 예방 및 치료뿐 아니라 피부 노화의 예방 관점에서 클로로필의 효능을 검토하고 확인했다는 것에서 의미가 있다. 그러나 본 연구 결과만으로는 클로로필의 농도 의존적 효능에 대한 명확한 판단을 하기에 부족한 부분이 있어 클로로필 효능의 농도 의존성에 대한 검토가 필요할 것으로 보여진다. 클로로필의 농도에 따른 효능 및 세포 독성의 상관성을 파악하고 동물, 인체효능 용량에 대한 연구를 통해 가장 바람직한 효능을 나타내는 클로로필의 농도를 산정하기 위한 검토가 이루어져야 할 것이다. 또한 피부 노화 예방을 위한 유용한 소재로서 클로로필의 임상적 활용 가능성을 판단하기 위해서는 동물, 인체 시험을 통한 클로로필의 효능 검증이 체계적으로 진행되어야 할 것으로 보여진다.

Keywords

References

  1. Chernomorsky SA, Segelman AB. Biological activities of chlorophyll derivatives. N. J. Med. 85: 669-673 (1988)
  2. Rudolph C. The therapeutic value of chlorophyll. Clin. Med. Surg. 37:119-121 (1930)
  3. de Vogel J, Jonker-Termont DS, van Lieshout EM, Katan MB, van der Meer R. Green vegetables, red meat and colon cancer: Chlorophyll prevents the cytotoxic and hyperproliferative effects of haem in rat colon. Carcinogenesis 26: 387-393 (2005) https://doi.org/10.1093/carcin/bgh331
  4. McCarty MF. The chlorophyll metabolite phytanic acid is a natural rexinoid--potential for treatment and prevention of diabetes. Med. Hypotheses 56: 217-219 (2001) https://doi.org/10.1054/mehy.2000.1153
  5. Okai Y, Higashi-Okai K, Yano Y, Otani S. Suppressive effects of chlorophyllin on mutagen-induced umu C gene expression in Salmonella typhimurium (TA 1535/pSK 1002) and tumor promoterdepenendent ornithine decarboxylase induction in BALB/c 3T3 fibroblast cells. Mutat. Res. 370: 11-17 (1996) https://doi.org/10.1016/S0165-1218(96)90122-6
  6. Higashi-Okai K, Okai Y. Potent suppressive activity of chlorophyll a and b from green tea (Camellia sinensis) against tumor promotion in mouse skin. J. UOEH. 20: 181-188 (1998)
  7. Kim J, Yook JI, Park KK, Jung SY, Hong JC, Kim KJ, Kim JA, Chung WY. Anti-promotion effect of chlorophyllin in DMBATPA- induced mouse skin carcinogenesis. Anticancer Res. 20: 1493-1498 (2000)
  8. Nakamura Y, Murakami A, Koshimizu K, Ohigashi H. Inhibitory effect of pheophorbide a, a chlorophyll-related compound, on skin tumor promotion in ICR mouse. Cancer Lett. 108: 247-255 (1996) https://doi.org/10.1016/S0304-3835(96)04422-9
  9. Cope RB, Loehr C, Dashwood R, Kerkvliet NI. Ultraviolet radiation- induced non-melanoma skin cancer in the Crl:SKH1:hr-BR hairless mouse: augmentation of tumor multiplicity by chlorophyllin and protection by indole-3-carbinol. Photochem. Photobio. Sci. 5: 499-507 (2006) https://doi.org/10.1039/b515556h
  10. Chug WY, Lee JM, Park MY, Yook JI, Kim J, Chung AS, Surh YJ, Park KK. Inhibitory effects of chlorophyllin on 7,12-dimethylbenz[a]anthracene-induced bacterial mutagenesis and mouse skin carcinogenesis. Cancer Lett. 14:5 57-64 (1999) https://doi.org/10.1016/S0304-3835(99)00229-3
  11. Kamat JP, Boloor KK, Devasagayam TP. Chlorophyllin as an effective antioxidant against membrane damage in vitro and ex vivo. Biochim. Biophys. Acta 1487: 113-127 (2000) https://doi.org/10.1016/S1388-1981(00)00088-3
  12. Robinson JP, Bruner LH, Bassoe CF, Hudson JL, Ward PA, Phan SH. Measurement of intracellular fluorescence of human monocytes relative to oxidative metabolism. J. Leukocyte Biol. 43: 304-310 (1988)
  13. Jiang SJ, Chu AW, Lu ZF, Pan MH, Che DF, Zhou XJ. Ultraviolet B-induced alterations of the skin barrier and epidermal calcium gradient. Photochem. Photobio. Sci. 5: 499–507 (2006) https://doi.org/10.1039/b515556h
  14. Ho JN, Cho HY, Lim EJ, Kim HK. Effects of aucbin isolated from Eucommia ulmoides on UVB-induced oxidative stress in human keratinocytes HaCaT. Food Sci. Biotechnol. 18: 475-480 (2009)
  15. Kong DS, Kim YA, Park JS, Kim SK, Lee BJ, Nam TJ, Seo Y. Antioxidant activity and inhibition of MMP-9 by isorhamnetin quercetin 3-O-b-glucopyranosides isolated from salicornia herbacea in HT 1080 cells. Food Sci. Biotechnol. 17: 983-989 (2008)
  16. Svobodov$\acute{a}$ A, Zdarilov$\acute{a}$ A, Walterov$\acute{a}$ D, Vost$\acute{a}$lov$\acute{a}$ J. Flavonolignans from Silybum marianum moderate UVA-induced oxidative damage to HaCaT keratinocytes. J. Dermatol. Sci. 48: 213-224 (2007) https://doi.org/10.1016/j.jdermsci.2007.06.008
  17. Moon HJ, Lee SR, Shim SN, Jeong SH, Stonik VA, Rasskazov VA, Zvyagintseva T, Lee YH. Fucoidan inhibits UVB-induced MMP-1 expression in human skin fibroblasts. Biol. Pharm. Bull. 31: 284-289 (2008) https://doi.org/10.1248/bpb.31.284
  18. Ritti$\acute{e}$ L, Fisher GJ. UV-light-induced signal cascades and skin aging. Ageing Res. Rev. 1: 705-720 (2002) https://doi.org/10.1016/S1568-1637(02)00024-7
  19. Yin ST, Tang ML, Deng HM, Xing TR, Chen JT, Wang HL, Ruan DY. Epigallocatechin-3-gallate induced primary cultures of rat hippocampal neurons death linked to calcium overload and oxidative stress. N-S Arch. Pharmacol. 379: 551-564 (2009) https://doi.org/10.1007/s00210-009-0401-4
  20. Isbrucker RA, Edwards JA, Wolz E, Davidovich A, Bausch J. Safety studies on epigallocatechin gallate (EGCG) preparations. Part 3: Teratogenicity and reproductive toxicity studies in rats. Food Chem. Toxicol. 44: 651-661 (2006) https://doi.org/10.1016/j.fct.2005.11.002
  21. Kobayashi T, Hattori S, Shinkai H. Matrix metalloproteinases-2 and -9 are secreted from human fibroblasts. Acta Derm. -Venereol. 83: 105-107 (2003) https://doi.org/10.1080/00015550310007436
  22. Overall CM, Wrana JL, Sodek J. Independent regulation of collagenase, 72-kDa progelatinase, and metalloendoproteinase inhibitor expression in human fibroblasts by transforming growth factor-b. J. Biol. Chem. 264: 1860 – 1869 (1989)
  23. Yang ES, Hong RH, Kang SM. The Effect of Genistein on the Proliferation and Type I pN Collagen Synthesis in Aged Normal Human Fibroblasts. Korean J. Microbiol. Biotechnol. 35: 316–324 (2007)
  24. Kang S, Chung JH, Lee JH, Fisher GJ, Wan YS, Duell EA, Voorhees JJ. Topical N-acetyl cysteine and genistein prevent ultraviolet-light-induced signaling that leads to photoaging in human skin in vivo. J. Invest. Dermatol. 120: 835-841 (2003) https://doi.org/10.1046/j.1523-1747.2003.12122.x