J Korean Med Sci.  2010 Jun;25(6):930-937. 10.3346/jkms.2010.25.6.930.

Prevention of UV-Induced Skin Damages by 11,14,17-Eicosatrienoic Acid in Hairless Mice In Vivo

• Jin XJ
• Kim EJ
• Oh IK
• Kim YK
• Park CH
• Chung JH

• KMID: 2150875
• DOI: http://doi.org/10.3346/jkms.2010.25.6.930

Abstract

Polyunsaturated fatty acids (PUFAs) are known to play important roles in various physiological and pathological processes. Recent studies have shown that some omega-3 (omega-3) PUFAs, such as eicosapentaenoic acid (EPA) and dodecahexaenoic acid (DHA), have protective effects on acute and chronic UV-induced changes. However, the effects of other omega-3 PUFAs including 11,14,17-eicosatrienoic acid (20:3) (ETA) on UV-induced skin damages are poorly understood. In this study, we investigated the cutaneous photoprotective effects of ETA in hairless mice in vivo. Female HR-1 hairless mice were topically treated with vehicle (ethanol:polyethylene glycol=30:70) only, 0.1% ETA, or 1% ETA once a day for 3 successive days after one time UV irradiation (200 mJ/cm2) on dorsal skins. Skin biopsy was carried out on the fourth day (72 hr after UV irradiation). We found that topical treatment with ETA attenuated UV-induced epidermal and dermal thickness and infiltration of inflammatory cells, and impairment of skin barrier function. In addition, ETA suppressed the expression of IL-1beta, COX-2, and MMP-13 induced by UV irradiation. Our results show that the topical application of ETA protects against UV-induced skin damage in hairless mice and suggest that ETA can be a potential agent for preventing and/or treating UV-induced inflammation and photoaging.

Keyword

Fatty Acids, Unsaturated; 11,14,17-eicosatrienoic acid; Skin Aging; Photoinflammation; Photoprotection; Ultraviolet Rays

Figure

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  Fig. 1 Topical application of 11,14,17-eicosatrienoic acid (ETA) inhibits UV-induced skin thickening and inflammatory cell infiltration in hairless mouse skin in vivo. Female HR-1 hairless mice were topically treated with vehicle (ethanol:polyethylene glycol=30:70) only, 0.1% ETA or 1% ETA once a day for 3 successive days after one time irradiation of UV (200 mJ/cm2) on dorsal skins. Skin biopsy was carried out on the fourth day (72 hr after UV irradiation). (A) Serial sections were mounted onto silane-coated slides and stained with hematoxylin and eosin (H&E) (magnification ×200). (B, C) Epidermal and dermal thicknesses were measured. Values are mean±SEM (n=8). *P<0.001 vs. control (CON) vehicle group; †P<0.05; ‡P<0.01 vs. UV-irradiated (UV) vehicle group.

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  Fig. 2 Topical application of ETA increases stratum corneum hydration and decreases transepidermal water loss (TEWL) in control and UV-irradiated hairless mouse skin in vivo. Female HR-1 hairless mice were topically treated with vehicle (ethanol:polyethylene glycol=30:70) only, 0.1% ETA or 1% ETA once a day for 3 successive days after one time irradiation of UV (200 mJ/cm2) on dorsal skins. Skin biopsy was carried out on the fourth day (72 hr after UV irradiation). (A) Stratum corneum hydration was dramatically decreased after UV irradiation. Stratum corneum hydration was increased by topical application of ETA both in normal and UV-irradiated hairless mice. (B) TEWL was dramatically increased after UV irradiation. Topical application of ETA decreased TEWL both in control and UV-irradiated hairless mice. Values are mean±SEM (n=8). *P<0.05; ‡P<0.001 vs. control (CON) vehicle group; ‡P<0.05 vs. UV-irradiated (UV) vehicle group.

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  Fig. 3 Topical application of ETA prevents UV-induced interleukin-1beta (IL-1β) and cyclooxygenase-2 (COX-2) expressions in hairless mouse skin in vivo. Female HR-1 hairless mice were topically treated with vehicle (ethanol:polyethylene glycol=30:70) only, 0.1% ETA or 1% ETA once a day for 3 successive days after one time irradiation of UV (200 mJ/cm2) on dorsal skins. Skin biopsy was carried out on the fourth day (72 hr after UV irradiation). (A) Expression of IL-1β mRNA was determined by quantitative real-time RT-PCR. (B) Expression of IL-1β protein was analyzed by ELISA. (C) Expression of COX-2 mRNA was detected by quantitative real-time RT-PCR. (D) Expression of COX-2 protein was detected by Western blot analysis. Results are expressed as fold change. Values are mean±SEM (n=8). *P<0.01, †P<0.001 vs. control (CON) vehicle group; ‡P<0.05, §P<0.01 ∥P<0.001 vs. UV-irradiated (UV) vehicle group.

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  Fig. 4 Topical application of ETA prevents UV-induced matrix metalloproteinase-13 (MMP-13) expression. Female HR-1 hairless mice were topically treated with vehicle (ethanol:polyethylene glycol=30:70) only, 0.1% ETA or 1% ETA once a day for 3 successive days after one time irradiation of UV (200 mJ/cm2) on dorsal skins. Skin biopsy was carried out on the fourth day (72 h after UV irradiation). (A) Expression of MMP-13 mRNA was detected by quantitative real-time RT-PCR. (B) Expression of MMP-13 protein was detected by western blot analysis. (C) Immunohistochemical staining of MMP-13 protein (magnification ×200). Results are expressed as fold change. Values are mean±SEM (n=8). *P<0.001 vs. control (CON) vehicle group; †P<0.05, ‡P<0.01 vs. UV-irradiated (UV) vehicle group.

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  Fig. 5 ETA increases IκBα expression and decreases NF-κB p65 phosphorylation in hairless mouse skin in vivo. Female HR-1 hairless mice were topically treated with vehicle (ethanol : polyethylene glycol=30:70) only, 0.1% ETA or 1% ETA once a day for 3 successive days after one time irradiation of UV (200 mJ/cm2) on dorsal skins. Skin biopsy was carried out on the fourth day (72 h after UV irradiation). (A) Expression of IκBα was detected by western blot analysis. (B) Phospho-NF-κB p65 was detected by western blot analysis. Results are expressed as fold change. Values are mean±SEM (n=8). *P<0.001 vs. control (CON) vehicle group; †P<0.05, ‡P<0.001 vs. UV-irradiated (UV) vehicle group.

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