To the best of our knowledge, no study has been conducted to date to directly compare the collagen metabolism of photoaged and naturally aged human skin. In this study, we compared collagen synthesis, matrix metalloproteinase-1 levels, and gelatinase activity of sun-exposed and sun-protected skin of both young and old subjects. Using northern blot analysis, immunohistochemical stain, and Western blot analysis, we demonstrated that the levels of procollagen type I mRNA and protein in photoaged and naturally aged human skin in vivo are significantly lower than those of young skin. Furthermore, we demonstrated, by northern blot analysis, that the procollagen alpha1(I) mRNA expression of photoaged skin is much greater than that of sun-protected skin in the same individual. In situ hybridization and immunohistochemical stain were used to show that the expression of type I procollagen mRNA and protein in the fibroblasts of photoaged skin is greater than for naturally aged skin. In addition, it was found, by Western blot analysis using protein extracted from the dermal tissues, that the level of procollagen type I protein in photoaged skin is lower than that of naturally aged skin. The level of matrix metalloproteinase-1 protein and the activity of matrix metalloproteinase-2 were higher in the dermis of photoaged skin than in naturally aged skin. Our results suggest that the natural aging process decreases collagen synthesis and increases the expression of matrix metalloproteinases, whereas photoaging results in an increase of collagen synthesis and greater matrix metalloproteinase expression in human skin in vivo. Thus, the balance between collagen synthesis and degradation leading to collagen deficiency is different in photoaged and naturally aged skin.
This is a comprehensive study of the changes in major antioxidant enzymes and antioxidant molecules during intrinsic aging and photoaging processes in the epidermis and dermis of human skin in vivo. We show that the activities of superoxide dismutase and glutathione peroxidase are not changed during these processes in human skin in vivo. Interestingly, the activity of catalase was significantly increased in the epidermis of photoaged (163%) and naturally aged (118%) skin (n = 9), but it was significantly lower in the dermis of photoaged (67% of the young skin level) and naturally aged (55%) skin compared with young (n = 7) skin. The activity of glutathione reductase was significantly higher (121%) in naturally aged epidermis. The concentration of alpha-tocopherol was significantly lower in the epidermis of photoaged (56% of young skin level) and aged (61%) skin, but this was not found to be the case in the dermis. Ascorbic acid levels were lower in both epidermis (69% and 61%) and dermis (63% and 70%) of photoaged and naturally aged skin, respectively. Gluta thione concentrations were also lower. Uric acid did not show any significant changes. Our results suggest that the components of the antioxidant defense system in human skin are probably regulated in a complex manner during the intrinsic aging and photoaging processes.
Human macrophage metalloelastase is a member of the matrix metalloproteinase family and is involved in degradation of elastin. We investigated the ultraviolet modulation of human macrophage metalloelastase in human skin in vivo. Ultraviolet induced human macrophage metalloelastase mRNA maximally (11.9-fold) within 16 h post-ultraviolet in human skin. This induction of human macrophage metalloelastase by ultraviolet was inhibited by pretreatment with the antioxidant N-acetyl cystein (20%) and vitamin E (5%) by an average of 54% and 47%, respectively, in human skin in vivo. Ultraviolet (30 mJ per cm2) and phorbol ester (12-O-tetradecanoyl-phorbol-13-acetate, 50 nM) treatment increased expression of human macrophage metalloelastase mRNA and protein in the cultured human dermal fibroblasts, but not in the keratinocytes. Chronically sun-exposed human skin expressed significant amounts of human macrophage metalloelastase protein, which colocalized with the material of solar elastosis, whereas there was little expression in sun-protected skin of the same individuals. This study demonstrates that ultraviolet irradiation increases human macrophage metalloelastase expression in human skin in vivo, possibly in macrophages and fibroblasts, and ultraviolet-induced expression of human macrophage metalloelastase can be inhibited by antioxidant (N-acetyl cystein and vitamin E) pretreatment. Association of human macrophage metalloelastase with elastotic material suggests that it may play an important role in the development of solar elastosis, the hallmark of sun-induced damage in human skin in vivo.
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