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Abstract
Purpose
Solar ultraviolet (UV) radiation causes inflammation and matrix metalloproteinase (MMP) overexpression and extracellular matrix depletion, leading to skin photoaging such as wrinkle formation, dryness, and sagging. Activation of MMP is influenced by various molecules such as reactive oxygen species (ROS), proinflammatory cytokines, and transient receptor potential vanilloid type (TRPV)-1, which are increased in UV-irradiated skin cells. Aralia elata (AE) ethanolic extract was reported to inhibit ROS generation caused by UVB-irradiation in keratinocytes. In this study, we investigated the photoprotective effect of AE ethanolic extract on UVB-irradiated human keratinocytes (HaCaT) and human dermal fibroblasts (HDF).
Methods
AE was freeze-dried, extracted in 70% ethanol, and concentrated. Skin cells were treated with AE extract for 24 h and then exposed to UVB (55 mJ/cm2). After 48 h of incubation, proinflammatory cytokines, MMP-1, type-1 procollagen, and TRPV-1 levels were measured by ELISA or Western blotting.
Results
Treatment with AE extract (100 µg/mL) significantly inhibited UVB-induced IL-6, IL-8, and PGE2 production in HaCaT by 25.6%, 5.3%, and 70.2%, respectively, and also inhibited elevation of MMP-1 and TRPV-1 caused by UVB irradiation by 20.0% and 41.9%, respectively (p < 0.05). In HDF, AE extract treatment significantly inhibited both elevation of MMP-1 and reduction of type-1 procollagen caused by UVB irradiation (p < 0.05). In addition, type-1 procollagen was elevated by AE extract treatment in normal HDFs (p < 0.05).
Figures and Tables
Fig. 1
Cell viability of HaCaT and HDF treated with Aralia elata (AE) extract. Cell viability of HaCaT and HDF were measured at 48 h treatment of AE extract. Results were shown as mean ± SD (n = 3). *Significantly different at p < 0.05 compared to the control
Fig. 2
Viability of HaCaT and HDF cells treated with Aralia elata (AE) extract and UVB. Cells were treated with AE extract for 24 h, exposed to UVB (55 mJ/cm2) and cell viability were measured at 48 h incubation. Results were shown as mean ± SD (n = 3). *Significantly different at p < 0.05 compared to UVB(-) control. NS: not significant
Fig. 3
Effect of Aralia elata (AE) extract on proinflammatory mediator secretion in UVB irradiated HaCaT cells. Cells were treated with AE extract for 24 h, exposed to UVB (55 mJ/cm2) and IL-6, IL-8 and PGE2 concentrations at 48 h incubation were measured in the culture medium by ELISA. Results were shown as mean ± SD (n = 3). Means sharing the same alphabet on the bar are not significantly different at p < 0.05 by ANOVA and Duncan's multiple range test.
Fig. 4
Effect of Aralia elata (AE) extract on TRPV-1 and MMP-1 protein levels in UVB-irradiated HaCaT cells. Cells were treated with AE extract for 24 h, exposed to UVB (55 mJ/cm2) and the culture medium and the cells were collected at 48 h incubation. Protein level of MMP-1 (medium), TRPV-1 and β-actin (cell) determined by western blotting (A) and quantified (B). Results were shown as mean ± SD (n = 4). Means sharing the same alphabet on the bar are not significantly different at p < 0.05 by ANOVA and Duncan's multiple range test.
Fig. 5
Effect of Aralia elata (AE) extract on MMP-1 and type I-procollagen protein levels in normal HDFs or UVB-irradiated HDFs. Cells were treated with AE extract for 24 h, exposed to UVB (55 mJ/cm2) and the culture medium and the cells were collected at 48 h incubation. The protein level of MMP-1 (medium), type I procollagen and β-actin (cell) determined by western blotting (A) and quantified (B). Results were shown as mean ± SD (n = 3~8). Means sharing the same alphabet on the bar are not significantly different at p < 0.05 by ANOVA and Duncan's multiple range test. NS: not significant
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