J Korean Med Sci.  2012 Mar;27(3):300-306. 10.3346/jkms.2012.27.3.300.

Acute UV Irradiation Increases Heparan Sulfate Proteoglycan Levels in Human Skin

• Jung JY
• Oh JH
• Kim YK
• Shin MH
• Lee D
• Chung JH

• KMID: 1793002
• DOI: http://doi.org/10.3346/jkms.2012.27.3.300

Abstract

Glycosaminoglycans are important structural components in the skin and exist as various proteoglycan forms, except hyaluronic acid. Heparan sulfate (HS), one of the glycosaminoglycans, is composed of repeated disaccharide units, which are glucuronic acids linked to an N-acetyl-glucosamine or its sulfated forms. To investigate acute ultraviolet (UV)-induced changes of HS and HS proteoglycans (HSPGs), changes in levels of HS and several HSPGs in male human buttock skin were examined by immunohistochemistry and real-time quantitative polymerase chain reaction (qPCR) after 2 minimal erythema doses (MED) of UV irradiation (each n = 4-7). HS staining revealed that 2 MED of UV irradiation increased its expression, and staining for perlecan, syndecan-1, syndecan-4, CD44v3, and CD44 showed that UV irradiation increased their protein levels. However, analysis by real-time qPCR showed that UV irradiation did not change mRNA levels of CD44 and agrin, and decreased perlecan and syndecan-4 mRNA levels, while increased syndecan-1 mRNA level. As HS-synthesizing or -degrading enzymes, exostosin-1 and heparanase mRNA levels were increased, but exostosin-2 was decreased by UV irradiation. UV-induced matrix metalloproteinase-1 expression was confirmed for proper experimental conditions. Acute UV irradiation increases HS and HSPG levels in human skin, but their increase may not be mediated through their transcriptional regulation.

Keyword

Acute UV Irradiation; Human Skin; Heparan Sulfate; Heparan Sulfate Proteoglycan

MeSH Terms

Adult
Agrin/genetics
Antigens, CD44/genetics
Base Sequence
DNA Primers/genetics
Gene Expression/radiation effects
Glucuronidase/genetics
Heparan Sulfate Proteoglycans/genetics/*metabolism
Heparitin Sulfate/metabolism
Humans
Male
Matrix Metalloproteinase 1/genetics
N-Acetylglucosaminyltransferases/genetics
RNA, Messenger/genetics/metabolism
Skin/*metabolism/*radiation effects
Skin Aging/genetics/physiology
Syndecan-1/genetics
Syndecan-4/genetics
Ultraviolet Rays/*adverse effects
Young Adult

Figure

• 

  Fig. 1 Immunohistochemistry of HS and HSPGs in human male buttock skin with acute UV irradiation. Healthy young Korean male buttock skin was irradiated with 2 MED of UV and skin specimens were obtained at 0, 24, 48, or 74 hr after UV irradiation by punch biopsy. Immunohistochemical staining (n = 4-7) for HS (A), perlecan (B), syndecan-1 (C), syndecan-4 (D), CD44v3 (E), CD44s (F), and MMP-1 (G) was performed as described in Materials and Methods.

• 

  Fig. 2 Changes in HSPG mRNA expression in human male buttock skin by acute UV irradiation. Healthy young Korean male buttock skin was irradiated with 2 MED of UV and the skin specimens were obtained at 0, 24, 48, or 74 hr after UV irradiation by punch biopsy. After separation of the epidermis from the dermis, total RNA was extracted from the epidermis tissues, and the mRNA expression changes of perlecan (A), syndecan-1 (B), syndecan-4 (C), CD44 (D), agrin (E), and MMP-1 (F) by acute UV irradiation were analyzed by real-time qPCR (n = 6), as described in Materials and Methods. Data were analyzed using the comparative Ct method, normalized to 36B4, and presented as mean relative fold changes ± standard deviation. Statistical analyses were performed using a Wilcoxon signed-rank test. P values of less than 0.05 were considered statistically significant. N.D., not detected.

• 

  Fig. 3 Changes in mRNA expression levels of HS synthesizing and degrading enzymes in human male buttock skin by acute UV irradiation. Healthy young Korean male buttock skin was irradiated with 2 MED of UV and the skin specimens were obtained at 0, 24, 48, or 74 hr after UV irradiation by punch biopsy. After separation of the epidermis from the dermis, total RNA was extracted from the epidermis tissues, and the mRNA expression changes of exostosin-1 (A), exostosin-2 (B), and heparanase (C) by acute UV irradiation were analyzed by real-time qPCR (n = 6), as described in Materials and Methods. Data were analyzed using the comparative Ct method, normalized to 36B4, and presented as mean relative fold changes ± standard deviation. Statistical analyses were performed using a Wilcoxon signed-rank test. P values of less than 0.05 were considered statistically significant.

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