Antioxidant Activity and Anti-wrinkle Effects of Aceriphyllum rossii Leaf Ethanol Extract

Abstract

We evaluated the antioxidant activity and anti-wrinkle effects of Aceriphyllum rossii leaf ethanol extract (ARLEE) in vitro using human dermal fibroblasts. The total polyphenol and flavonoid contents of ARLEE were 578.6 and 206.3 mg/g, respectively. At a concentration of 250 μg/mL, the electron-donating ability of ARLEE was 87.1%. In comparison with the vehicle, ARLEE treatment at 100 μg/mL significantly increased type I procollagen synthesis (p < 0.01) by 50.7%. In vitro ARLEE treatment (10 mg/mL) inhibited collagenase and elastase activity by 97.1% and 99.2%, respectively. Compared with the control, ascorbic acid treatment at 100 μg/mL significantly decreased matrix metalloproteinase (MMP)-1 protein expression (p < 0.01) by 37.0%. ARLEE treatment at 50 μg/mL significantly decreased MMP-1 protein expression (p < 0.01) by 46.1%. Ascorbic acid and ARLEE treatments at 100 μg/mL significantly decreased MMP-1 mRNA expression (p < 0.01) by 26.1% and 36.1%, respectively. From these results, we conclude that ARLEE has excellent antioxidant activity and even better anti-wrinkle effects than ascorbic acid in human dermal fibroblasts. These results suggest that ARLEE could be used in functional cosmetics for the prevention or alleviation of skin wrinkles induced by ultraviolet rays.

Keywords: Aceriphyllum rossii; Antioxidant activity; Collagenase activity; MMP-1; Type I procollagen.

Fig. 1.. Total polyphenol and flavonoid contents of Aceriphyllum rossii ethanol extracts. Values represent the mean ± SD of three independent measurements. ^a,b,cValues with different superscripts indicate significant differences (p < 0.05) for each polyphenol or flavonoid content, by ANOVA and Duncan’s multiple range tests.

Fig. 2.. Electron-donating ability of Aceriphyllum rossii ethanol extracts relative to the control ascorbic acid at the indicated concentrations. Each substance was evaluated on its ability to provide electrons to the free radical DPPH. AA: ascorbic acid, ARLEE: A. rossii leaf ethanol extract, ARSEE: A. rossii stem ethanol extract, ARREE: A. rossii root ethanol extract. Values represent the mean ± SD of three independent measurements. ^a,bValues with different superscripts indicate significant differences (p < 0.05) for each given concentration, by ANOVA and Duncan’s multiple range tests.

Fig. 3.. Collagenase activity inhibition of ARLEE relative to the control ascorbic acid. AA: ascorbic acid, ARLEE: A. rossii leaf ethanol extract. Values are the mean ± SD of three independent measurements. The value with an asterisk is significantly different from the AA group by Student’s t-test (*; p < 0.05, **; p < 0.01, ***; p < 0.001).

Fig. 4.. Elastase activity inhibition of ARLEE relative to the control ascorbic acid. AA: ascorbic acid, ARLEE: A. rossii leaf ethanol extract. Values are the mean ± SD of three independent measurements. The value with an asterisk is significantly different from the AA group by Student’s t-test (***; p < 0.001).

Fig. 5.. Effect of ARLEE on collagen production in human dermal fibroblasts. Cells were treated with the vehicle (V), 5 ng/mL TGF-β1 (PC), or A. rossii leaf ethanol extract (ARLEE) at the indicated concentrations, and the production of procollagen was measured by ELISA. Values are the mean ± SD of three independent measurements. The value with an asterisk is significantly different from the vehicle group by Student’s t-test (*; p < 0.05, **; p < 0.01).

Fig. 6.. Effect of ARLEE on MMP-1 protein expression in human dermal fibroblasts. (A) MMP-1 protein levels decreased upon treatment with A. rossii leaf ethanol extract (ARLEE) compared to UVA-irradiated control cells, as determined by western blotting. Expression was normalized to β-actin levels. (B) Quantification of MMP-1 protein expression in cells treated with vehicle (V), 6.3 J/cm² UVA radiation (C), 6.3 J/cm² UVA radiation + 100 μg/mL ascorbic acid (PC), or 6.3 J/cm² UVA radiation + ARLEE at the indicated concentrations. Values are the mean ± SD of three independent measurements. The value with an asterisk is significantly different from the control group by Student’s t-test (**; p<0.01).

Fig. 7.. Effect of ARLEE on MMP-1 mRNA expression in human dermal fibroblasts. (A) MMP-1 transcript levels decreased upon treatment with A. rossii leaf ethanol extract (ARLEE) in a dosedependent manner compared to UVA-irradiated control cells, as determined by RT-PCR. Expression was normalized to β-actin levels. (B) Quantification of MMP-1 transcript expression in cells treated with the vehicle (V), 6.3 J/cm² UVA radiation (C), 6.3 J/cm² UVA radiation + 100 μg/mL ascorbic acid (PC), or 6.3 J/cm² UVA radiation + ARLEE at the indicated concentrations. Values are the mean ± SD of three independent measurements. The value with an asterisk is significantly different from the control group by Student’s t-test (*; p < 0.05, **; p < 0.01).