Dahae Lim1, Min Kyoung Kim2, Young-Pyo Jang3, Jinju Kim4. 1. Department of Korean Physiology, College of Pharmacy, Kyung Hee University, Seoul, South Korea. 2. Department of Life and Nanopharmaceutical Sciences, College of Pharmacy, Kyung Hee University, Hoegi-dong, Dongdaemun-gu, Seoul 130-701, South Korea. 3. Department of Life and Nanopharmaceutical Sciences, College of Pharmacy, Kyung Hee University, Hoegi-dong, Dongdaemun-gu, Seoul 130-701, South Korea; Department of Oriental Pharmaceutical Science, College of Pharmacy, Kyung Hee University, Hoegi-dong, Dongdaemun-gu, Seoul 130-701, South Korea. 4. Department of Korean Physiology, College of Pharmacy, Kyung Hee University, Seoul, South Korea. Electronic address: shdwer@khu.ac.kr.
Abstract
BACKGROUND: Sceptridium ternatum (ST) is a medicinal herb used in folk remedies for the treatment of various disorders such as pertussis, allergic asthma, abdominalgia, diarrhea, and external use for wound healing. However, the biological and pharmacological activities of ST are not fully clarified besides anti-asthmatic effect. OBJECTIVE: We studied a Sceptridium ternatum ethanol extract (ST) with respect to its anti-inflammatory and immune regulatory activities in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, concanavalin A (conA)-stimulated BALB/c mice splenocytes, and a 2,4-dinitrochlorobenzene (DNCB)-induced allergic contact dermatitis (ACD) mouse model. METHODS: RAW 264.7 cells were pretreated with ST for 1h and then stimulated with LPS. To determine the anti-inflammatory effects of ST, the production of nitric oxide (NO), interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) were measured using an enzyme-linked immunosorbent assay (ELISA). To determine its anti-allergic effects, splenocytes from BALB/c mice were incubated and stimulated with conA in the absence or presence of ST for 48h. The production of IL-4 and interferon (IFN)-γ in culture supernatants were evaluated by ELISA. To test the effects of ST on ACD, 100μL of 1% DNCB was applied to the dorsal skin of BALB/c mice for 2 weeks, and ST was administered 2 h before DNCB application. The thicknesses of the epidermis and dermis were determined by skin histological analysis. Serum immunoglobulin (Ig) E levels, the production of IL-1β, IL-4, and IL-6 in dorsal skin tissue, and T helper (Th) 2 cytokines production of CD4(+) T cells were analyzed by ELISA. The expression of nuclear transcription factor-κB (NF-κB) both in vitro and in vivo was determined via immunoblotting. RESULTS: In RAW 264.7 cells, ST inhibited LPS-induced inflammation mediator production and NF-κB expression. ST upregulated IFN-γ production and downregulated IL-4 production in conA-stimulated splenocytes. ST application reduced the thicknesses of the epidermis and dermis by decreasing serum IgE level and the expressions of IL-1β, IL-4, IL-6, and NF-κB in the dorsal skin of the DNCB-induced ACD model mice. Furthermore, ST treated group showed reduction of the Th2 cytokines production in activated CD4(+) T cells. CONCLUSION: These findings not only indicate that application of ST reduced skin thickening by regulating Th 2-type allergic responses and inhibiting expression of inflammatory mediators in a DNCB-induced ACD mouse model, but also suggest that Sceptridium ternatum is a natural option for the treatment of skin inflammation.
BACKGROUND:Sceptridium ternatum (ST) is a medicinal herb used in folk remedies for the treatment of various disorders such as pertussis, allergic asthma, abdominalgia, diarrhea, and external use for wound healing. However, the biological and pharmacological activities of ST are not fully clarified besides anti-asthmatic effect. OBJECTIVE: We studied a Sceptridium ternatumethanol extract (ST) with respect to its anti-inflammatory and immune regulatory activities in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, concanavalin A (conA)-stimulated BALB/c mice splenocytes, and a 2,4-dinitrochlorobenzene (DNCB)-induced allergic contact dermatitis (ACD) mouse model. METHODS: RAW 264.7 cells were pretreated with ST for 1h and then stimulated with LPS. To determine the anti-inflammatory effects of ST, the production of nitric oxide (NO), interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) were measured using an enzyme-linked immunosorbent assay (ELISA). To determine its anti-allergic effects, splenocytes from BALB/c mice were incubated and stimulated with conA in the absence or presence of ST for 48h. The production of IL-4 and interferon (IFN)-γ in culture supernatants were evaluated by ELISA. To test the effects of ST on ACD, 100μL of 1% DNCB was applied to the dorsal skin of BALB/c mice for 2 weeks, and ST was administered 2 h before DNCB application. The thicknesses of the epidermis and dermis were determined by skin histological analysis. Serum immunoglobulin (Ig) E levels, the production of IL-1β, IL-4, and IL-6 in dorsal skin tissue, and T helper (Th) 2 cytokines production of CD4(+) T cells were analyzed by ELISA. The expression of nuclear transcription factor-κB (NF-κB) both in vitro and in vivo was determined via immunoblotting. RESULTS: In RAW 264.7 cells, ST inhibited LPS-induced inflammation mediator production and NF-κB expression. ST upregulated IFN-γ production and downregulated IL-4 production in conA-stimulated splenocytes. ST application reduced the thicknesses of the epidermis and dermis by decreasing serum IgE level and the expressions of IL-1β, IL-4, IL-6, and NF-κB in the dorsal skin of the DNCB-induced ACD model mice. Furthermore, ST treated group showed reduction of the Th2 cytokines production in activated CD4(+) T cells. CONCLUSION: These findings not only indicate that application of ST reduced skin thickening by regulating Th 2-type allergic responses and inhibiting expression of inflammatory mediators in a DNCB-induced ACD mouse model, but also suggest that Sceptridium ternatum is a natural option for the treatment of skin inflammation.