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, 28 (4), 241-8

Effect of Microalgal Extracts of Tetraselmis Suecica Against UVB-Induced Photoaging in Human Skin Fibroblasts

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Effect of Microalgal Extracts of Tetraselmis Suecica Against UVB-Induced Photoaging in Human Skin Fibroblasts

Wol Soon Jo et al. Toxicol Res.

Abstract

Exposure of cells to ultraviolet B (UVB) radiation can induce production of free radicals and reactive oxygen species (ROS), which damage cellular components. In addition, these agents can stimulate the expression of matrix metalloproteinase (MMP) and decrease collagen synthesis in human skin cells. In this study, we examined the anti-photoaging effects of extracts of Tetraselmis suecica (W-TS). W-TS showed the strongest scavenging activity against 2,2-difenyl-1-picrylhydrazyl (DPPH) and peroxyl radicals, followed by superoxide anions from the xanthine/xanthine oxidase system. We observed that the levels of both intracellular ROS and lipid peroxidation significantly increased in UVB-irradiated human skin fibroblast cells. Furthermore, the activities of enzymatic antioxidants (e.g., superoxide dismutase) and the levels of non-enzymatic antioxidants (e.g., glutathione) significantly decreased in cells. However, W-TS pretreatment, at the maximum tested concentration, significantly decreased intracellular ROS and malondialdehyde (MDA) levels, and increased superoxide dismutase and glutathione levels in the cells. At this same concentration, W-TS did not show cytotoxicity. Type 1 procollagen and MMP-1 released were quantified using RT-PCR techniques. The results showed that W-TS protected type 1 procollagen against UVBinduced depletion in fibroblast cells in a dose-dependent manner via inhibition of UVB-induced MMP-1. Taken together, the results of the study suggest that W-TS effectively inhibits UVB-induced photoaging in skin fibroblasts by its strong anti-oxidant ability.

Keywords: Anti-oxidant; Photo aging; Reactive oxygen species; Tetraselmis suecica; Ultraviolet B.

Figures

Fig. 1.
Fig. 1.. Procedures for the preparation of crude extracts and fractions from Tetraselmis suecica. The extractive procedures described used water and enzyme digestion.
Fig. 2.
Fig. 2.. Cell viability of fibroblast cell with W-TS and generation of ROS in UVB treated human fibroblast cells treated with W-TS from Tetraselmis suecica. (A) Fibroblast cells were treated with various concentrations of the crude extracts (aqueous extract, W-TS) from Tetraselmis suecica for 24 hr. Cell viability was determined by MTT assay and expressed as a percentage of control growing cells. (B) Effect of W-TS on intracellular ROS in fibroblasts following UVB exposure. Fibroblast cells were pretreated with W-TS (31, 63, 125, 250 and 500 μg/ml) prior to UVB irradiation (100 mJ/cm2). Three independent assays were performed in triplicate and the data shown are mean ± S.D. *p< 0.05 vs. media alone and #p<0.05 vs. UVB induced group (control); significance of difference between the treated groups by Studentst tests (*) and ANOVA followed by Dunnett’s test (#).
Fig. 3.
Fig. 3.. The effect of W-TS from Tetraselmis suecica on the SOD, GSH and MDA levels in UVB irradiated fibroblasts. Fibroblast cells were pretreated with W-TS (31, 63, 125, 250 and 500 μg/ ml) prior to UVB irradiation (100 mJ/cm2) and harvested 24 hr later. Ascorbic acid was used as a positive control. (A) The SOD activity was measured using the SOD assay kit, as described in Materials and Methods. (B) The GSH contents were measured using the DTNB, as described in Materials and Methods. (C) The MDA was estimated with MDA-586 kit and spectrometer at 586 nm. Three independent assays were performed in triplicate and the data shown are mean ± S.D. *p< 0.05 vs. media alone and #p< 0.05 vs. UVB induced group (control); significance of difference between the treated groups by Students-t tests (*) and ANOVA followed by Dunnett’s test (#).
Fig. 4.
Fig. 4.. The effect of W-TS from Tetraselmis suecica on the UVB-induced MMP-1 and type I procollagen mRNA production in human skin fibroblasts. Cells were pretreated with W-TS (31, 63, 125, 250 and 500 μg/ml) prior to UVB irradiation (100 mJ/cm2) and harvested 24 h later. MMP-1 (A) and type 1 procollagen mRNA production (B) was determined by RT-PCR. MMP-1 and type 1 procollagen mRNA was normalized vs. that of the corresponding GAPDH mRNA. Ascorbic acid was used as a positive control. Three independent assays were performed in triplicate and the data shown are mean ± S.D. *p< 0.05 vs. media alone and #p< 0.05 vs. UVB induced group (control); significance of difference between the treated groups by Students-t tests (*) and ANOVA followed by Dunnett’s test (#).

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