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The effects of ultraviolet A and reactive oxygen species on the mRNA expression of 72-kDa type IV collagenase and its tissue inhibitor in cultured human dermal fibroblasts

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Abstract

The effects of ultraviolet A (UVA) radiation and reactive oxygen species (ROS), generated with a xanthine and xanthine oxidase (XOD) system, on collagen enzymatic degradation involving the matrix metalloproteinase (MMP) and its tissue inhibitor of metalloproteinase (TIMP) were investigated using cultured human dermal fibroblasts. Total RNA was isolated and subjected to Northern blot analysis using cDNA clones for human interstitial collagenase (MMP-1), 72-kDa type IV collagenase (MMP-2) and TIMP-2. UVA irradiation resulted in an increase in MMP-1 mRNA up to 2.3-fold, but did not stimulate MMP-2 or TIMP-2 mRNA expression. In contrast, ROS induced by the xanthine and XOD system resulted in a dose-related increase in the level of MMP-2 mRNA up to 2.1-fold and a decrease in the level of TIMP-2 mRNA by 49% in the same fibroblasts. Catalase, used as scavenger, essentially prevented the ROS-induced alterations in MMP-2 and TIMP-2 mRNA levels. These results suggest that ROS produced in the dermis may contribute to biological changes in the connective tissue matrix observed in photoaging skin by accelerating the MMP-2-related matrix degradation system.

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References

  1. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  2. Brenner DA, O'Hara M, Angel P, Chojkier M, Karin M (1989) Prolonged activation of jun and collagenase genes by tumor necrosis factor-alpha. Nature 337:661–663

    Article  PubMed  CAS  Google Scholar 

  3. Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanat-phenol-chloroform extraction. Anal Biochem 162:156–159

    Article  PubMed  CAS  Google Scholar 

  4. Collier IE, Wilhelm SM, Eisen AZ, Marmer BL, Grant GA, Seltzer JL, Kronberger A, He C, Bauer EA, Goldberg GI (1988) H-ras oncogene-transformed human bronchial epithelial cells (TBE-1) secrete a single metalloproteinase capable of degrading basement membrane collagen. J Biol Chem 263:6579–6587

    PubMed  CAS  Google Scholar 

  5. Crawford D, Zbinden I, Amstrad P, Cerutti P (1988) Oxidant stress induces the proto-oncogenes c-fos and c-myc in mouse epidermal cells. Oncogene 3:27–32

    CAS  Google Scholar 

  6. Docherty AJP, Lyons A, Smith BJ, Wright EM, Stephens PE, Harris TJR, Murphy G, Reynolds JJ (1985) Sequence of human tissue inhibitor of metalloproteinases and its identity to erythroid-potentiating activity. Nature 318:66–69

    Article  PubMed  CAS  Google Scholar 

  7. Gross J, Lapiere CM (1962) Collagenolytic activity in amphibian tissues: a tissue culture assay. Proc Natl Acad Sci USA 48: 1014–1022

    Article  PubMed  CAS  Google Scholar 

  8. Howard EW, Bullen EC, Banda MJ (1991) Preferential inhibition of 72- and 92-kDA gelatinases by tissue inhibitor of metalloproteinases-2. J Biol Chem 266:13070–13075

    PubMed  CAS  Google Scholar 

  9. Kligman AM (1969) Early destructive effects of sunlight on human skin. J Am Med Assoc 210: 2377–2380

    Article  CAS  Google Scholar 

  10. Kligman LH, Kligman AM (1986) The nature of photoaging: its prevention and repair. Photodermatol 3:215–227

    PubMed  CAS  Google Scholar 

  11. Kligman LH, Gebre M, Alper R, Kefalides NA (1989) Collagen metabolism in ultraviolet irradiated hairless mouse skin and its correlation to histochemical observations. J Invest Dermatol 93:210–214

    Article  PubMed  CAS  Google Scholar 

  12. Kligman LH, Akin FJ, Kligman AM (1992) Prevention of ultraviolet damage to the dermis of hairless mice by sunscreens. J Invest Dermatol 78:181–189

    Article  Google Scholar 

  13. Kucharz EJ (1992) The collagens: biochemistry and pathophysiology, chapter 4. Degradation. Springer, Berlin Heidelberg New York, pp 55–67

    Google Scholar 

  14. Lavker RM (1979) Structural alterations in exposed and unexposed aged skin. J Invest Dermatol 73: 59–66

    Article  PubMed  CAS  Google Scholar 

  15. Lavker RM, Kligman AM (1988) Chronic heliodermatitis: a morphologic evaluation of chronic actinic dermal damage with emphasis on the role of mast cells. J Invest Dermatol 90:325–330

    Article  PubMed  CAS  Google Scholar 

  16. Maki A, Berezesky IK, Fargnoli J, Holbrook NJ, Trump BF (1992) Role of [Ca2+]i in induction of c-fos, c-jun and c-myc mRNA in rat PTE after oxidative stress. FASEB J 6:919–924

    PubMed  CAS  Google Scholar 

  17. Murphy G, McAlpine CG, Poll CT, Reynolds JJ (1985) Purification and characterization of a bone metalloproteinase that degrades gelatin and types IV and V collagen. Biochem Biophys Acta 831:49–58

    PubMed  CAS  Google Scholar 

  18. Naruo S, Kanayama H, Aki M, Kagawa S (1993) Gene expressions of type IV collagenase and tissue inhibitor of metalloproteinase (TIMP) in human bladder cancers. Nippon-Hinyokika-Gakkai-Zasshi 84:841–850

    PubMed  CAS  Google Scholar 

  19. Oikarinen A, Karvonen J, Uitto J, Hannuksela M (1985) Connective tissue alterations in skin exposed to natural and therapeutic UV-radiation. Photodermatology 2:115–126

    Google Scholar 

  20. Overall CM, Wrana JL, Sodek J (1991) Transcriptional and post-transcriptional regulation of 72-kDa gelatinase/type IV collagenase by transforming growth factor-beta 1 in human fibroblasts. J Biol Chem 266:14064–14071

    PubMed  CAS  Google Scholar 

  21. Petersen MJ, Hansen C, Craig S (1992) Ultraviolet A irradiation stimulates collagenase production in cultured human fibroblasts. J Invest Dermatol 99:440–444

    Article  PubMed  CAS  Google Scholar 

  22. Sato H, Kida Y, Mai M, Endo Y, Sasaki T, Tanaka J, Seiki M (1992) Expression of genes encoding type IV collagen-degrading metalloproteinases and tissue inhibitors of metalloproteinases in various human tumor cells. Oncogene 77:77–83

    Google Scholar 

  23. Scharffetter-Kochanek K, Wlaschek M, Hogg A, Bolsen K, Schothorst A, Goerz G, Krieg T, Plewig G (1991) UVA irradiation induces collagenase in human dermal fibroblasts in vitro and in vivo. Arch Dermatol Res 283:506–511

    Article  Google Scholar 

  24. Scharffetter-Kochanek K, Wlaschek M, Briviba K, Sies H (1993) Singlet oxygen induces collagenase expression in human skin fibroblasts. FEBS Lett 331:304–306

    Article  PubMed  CAS  Google Scholar 

  25. Schonthal A, Herrlich P, Rahmsdorf HJ, Ponta H (1988) Requirement for fos gene expression in the transcriptional activation of collagenase by other oncogenes and phorbol esters. Cell 54:325–334

    Article  PubMed  CAS  Google Scholar 

  26. Seltzer JL, Eisen AZ, Bauer EA, Morris NP, Glanville RW, Burgeson RE (1989) Cleavage of type VII collagen by interstitial collagenase and type IV collagenase (gelatinase) derived from human skin. J Biol Chem 264:3822–3826

    PubMed  CAS  Google Scholar 

  27. Shimizu S, Malik K, Sejima H, Kishi J, Hayakawa T, Koiwai O (1992) Cloning and sequencing of the cDNA encoding a mouse tissue inhibitor of metalloproteinase-2. Gene 114:291–292

    Article  PubMed  CAS  Google Scholar 

  28. Smith JG, Davidson EA, Sams WM, Clark RD (1962) Alterations in human dermal connective tissue with age and chronic sun damage. J Invest Dermatol 39:347–350

    Article  PubMed  CAS  Google Scholar 

  29. Stetler-Stevenson WG, Krutzsch HC, Liotta LA (1989) Tissue inhibitor of metalloproteinase (TIMP-2). J Biol Chem 264: 17374–17378

    PubMed  CAS  Google Scholar 

  30. Sugiura K, Ueda H, Hirano K, Adachi T (1985) Studies on superoxide dismutase in human skin. (2) Contents of superoxide dismutase and lipid peroxide in normal human skin. Jpn J Dermatol 95:1541–1545

    CAS  Google Scholar 

  31. Tanaka H, Okada T, Konishi H, Tsuji T (1993) The effect of reactive oxygen species on the biosynthesis of collagen and glycosaminoglycans in cultured human dermal fibroblasts. Arch Dermatol Res 285:352–355

    Article  PubMed  CAS  Google Scholar 

  32. Trautinger F, Gruenwald C, Trenz A, Pittermann W, Kokoschka EM (1989) Influence of natural vitamin E and UV radiation on dermal collagen content in hairless mice. Arch Dermatol Res 283:39

    Google Scholar 

  33. Warren R, Gartstein V, Kligman AM, Montagna W, Allendorf RA, Ridder GM (1991) Age, sunlight, and facial skin: a histologic and quantitative study. J Am Acad Dermatol 25:751–760

    Article  PubMed  CAS  Google Scholar 

  34. Wlaschek M, Bolsen K, Herrmann G, Schwarz A, Wilmroth F, Heinrich PC, Goerz G, Scharffetter-Kochanek K (1993) UFA-induced autocrine stimulation of fibroblast-derived-collagenase by IL-6: a possible mechanism in dermal photodamage? J Invest Dermatol 101:164–168

    Article  PubMed  CAS  Google Scholar 

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Kawaguchi, Y., Tanaka, H., Okada, T. et al. The effects of ultraviolet A and reactive oxygen species on the mRNA expression of 72-kDa type IV collagenase and its tissue inhibitor in cultured human dermal fibroblasts. Arch Dermatol Res 288, 39–44 (1996). https://doi.org/10.1007/BF02505041

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  • DOI: https://doi.org/10.1007/BF02505041

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