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. 2016:2016:8027537.
doi: 10.1155/2016/8027537. Epub 2016 Aug 28.

Anti-Inflammatory Effects of Artemisia Leaf Extract in Mice with Contact Dermatitis In Vitro and In Vivo

Chanyong Yun  1 , Youngchul Jung  1 , Wonjoo Chun  1 , Beodeul Yang  1 , Junghyun Ryu  1 , Chiyeon Lim  2 , Jung-Hoon Kim  1 , Hyungwoo Kim  1 , Su-In Cho  1
Affiliations

Anti-Inflammatory Effects of Artemisia Leaf Extract in Mice with Contact Dermatitis In Vitro and In Vivo

Chanyong Yun et al. Mediators Inflamm. 2016.

Abstract

The leaves of Artemisia argyi Lev. et Vant. and A. princeps Pamp. are well known medicinal herbs used to treat patients in China, Japan, and Korea with skin problems such as eczema and itching, as well as abdominal pain and dysmenorrhoea. We investigated the anti-inflammatory effects of Artemisia leaf extract (ALE) using CD mice and Raw 264.7 cells. The effects of ALE on histopathological changes and cytokine production in ear tissues were assessed in mice with CD induced by 1-fluoro-2,4-dinitrobenzene (DNFB). Moreover, the anti-inflammatory effects on production levels of prostaglandin E2 (PGE2) and nitric oxide (NO) and expression levels of cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) were investigated in Raw 264.7 cells. Topical application of ALE effectively prevented ear swelling induced by repeated DNFB application. ALE prevented epidermal hyperplasia and infiltration of immune cells and lowered the production of interferon- (IFN-) gamma (γ), tumour necrosis factor- (TNF-) alpha (α), and interleukin- (IL-) 6 in inflamed tissues. In addition, ALE inhibited expression of COX-2 and iNOS and production of NO and PGE2 in Raw 264.7 cells. These results indicate that Artemisia leaf can be used as a therapeutic agent for inflammatory skin diseases and that its anti-inflammatory effects are closely related to the inhibition of inflammatory mediator release from macrophages and inflammatory cytokine production in inflamed tissues.

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Figures

Figure 1
Figure 1
Experimental schedule. Mice in all experimental groups were sensitized by DNFB for three successive days and then challenged every other day (four times). The ALE group was topically treated with 30 or 300 μg/ear of ALE (n = 8). The DEX group was topically treated with 75 μg/ear of DEX for seven days from day 8 to day 14 (n = 6). All animals were sacrificed on day 15. S indicates sacrifice.
Figure 2
Figure 2
Effect of ALE on ear swelling in CD mice. The ear thickness and weight were measured on day 15. NOR: nontreated normal; CTL: nontreated CD; 30 or 300, 30, or 300 μg/ear of ALE; DEX: 75 μg/ear of DEX. (a) Ear thickness. (b) Ear weight. Values are presented as the means ± SD. ## P < 0.01 and ### P < 0.001 compared to the NOR group and ∗ P < 0.05 and ∗∗ P < 0.01 compared to the CTL group.
Figure 3
Figure 3
Effect of ALE on epidermal hyperplasia and infiltration of immune cells in CD mice. Skin tissue was observed under a light microscope. (A) NOR; ((B) and (C)) CTL; (D) 30 μg/ear of ALE; (E) 300 μg/ear of ALE; (F) 75 μg/ear of DEX. Magnification, 50x. Yellow bars indicate the epidermis. Filled arrows indicate pustule areas (a). The epidermal thickness (A) and infiltration of immune cells (B) were evaluated using a quantitative method. Abbreviations are the same as in Figure 2. (b). Values are presented as the means ± SD. ## P < 0.01 compared to the NOR group and ∗ P < 0.05 compared to the CTL group.
Figure 4
Figure 4
Effect of ALE on production levels of cytokines in inflamed tissues. The cytokine levels in inflamed tissues were evaluated. (a) TNF-α; (b) IFN-γ; (c) IL-6; (d) IL-10. N.D., not detected. Values are presented as the means ± SD. # P < 0.05 and ### P < 0.001 compared to the NOR group and ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 compared to the CTL group.
Figure 5
Figure 5
Effects of ALE on iNOS expression and NO production in Raw 264.7 cells. Cells were incubated with various concentrations of ALE for 4 h and then activated with 1 μg/mL of LPS for 20 h. (a) iNOS expression; (b) NO production. Values are presented as the means ± SD. ## P < 0.01 compared to the NOR group and ∗ P < 0.05 and ∗∗ P < 0.01 compared to the CTL group.
Figure 6
Figure 6
Effects of ALE on COX-2 expression and PGE2 production in Raw 264.7 cells. Cells were treated with various concentrations of ALE for 4 h and then activated with 1 μg/mL of LPS for 20 h. The COX-2 expression levels were then detected by western blot analysis (a), while PGE2 production was measured by ELISA. Values represented are the means ± SD. ## P < 0.01 compared to the NOR group and ∗ P < 0.05 compared to the CTL group (b).
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