## 3 Results and Discussion
### Skin Antiging Effects of the Samples on UV\({}_{b}\)-Irradiated Human Fibroblasts  
As the results shown in Figure 6, proved that AF protected against mitochondrial dysfunction, further experiments are needed to confirm whether the increase in mitochondrial activities induced by AF affects the antiaging process of human skin. With increasing age, a sustained reduction in collagen levels and increased secretion of matrix-degrading enzymes such as matrix metalloproteinases (MMPs) in old skin were observed compared with young skin [45,51]. Therefore, Figures 7 and 8 show the inhibition of MMP-1 and the production of collagen along with the regulation of gene expression in UVb-irradiated human skin fibroblasts. The production of MMP-1, which is also an indicator of skin wrinkling processes, was observed to more clearly prove that the AF is involved in the skin antiaging process, as shown in Figure 7. In Figure 7, after irradiation with UVb light, MMP-1 levels were substantially increased up to 143% of the control cells (not exposed to UVb light) in which 125 ng/mL MMP-1 was produced (data not shown), while MMP-1 levels were decreased to 70.57 \(\pm\) 3.49% in UVb-irradiated cells with 10 \(\upmu\)g/mL quercetin. Treatments with 0.3% and 0.5% of the AW also resulted in a 75% reduction in MMP-1 production. Compared to those results, the AF significantly reduced MMP-1 production down to 69.41 \(\pm\) 2.63% at a dose of 0.3% without a further decrease in the presence of 0.5% of the AF. As shown in Figure 8a, skin fibroblast cells with mitochondrial damage induced by UVb light exhibited significantly reduced collagen production to 45% of the 45 ng/mL collagen produced by the control cells (data not shown). Quercetin alone enhanced the restoration of collagen production, up to 112.37 \(\pm\) 8.01% of the control, after treatment with 10 \(\upmu\)g/mL quercetin, and the addition of 0.3% of the AW induced a restoration effect similar to that of quercetin itself. Moreover, a 125 \(\pm\) 5.45% increase in collagen production was measured after treatment with 0.3% of the AF. Compared to the control, a value was much higher than that induced by quercetin itself with the same contents in the AF. This result clearly implies a synergistic effect of quercetin and other biological components in the AF. A similar restoration of collagen production was also observed in cells treated with 0.3% and 0.5% of the AW and AF. Thus, a saturation concentration may exist after which the extracts are no longer more effective at certain dosages, which in this case was 0.5%. However, the AF has the ability to prevent skin wrinkling processes, specifically in skin photo-aging, possibly by reversing mitochondrial dysfunction. The expression levels of the genes related to collagen production are also shown in Figure 8b,c. As shown in Figure 8b, the expression of collagen genes was substantially upregulated in UVb-irradiated cells treated with 0.3% of the AF as the concentration dependency manner (ColA1). Its band intensity was also digitized in Figure 8c for easy comparison with other bands, showing that all of the results were similar patterns to those presented in Figure 8b. Overall speaking, the protective effects of the AF on skin wrinkling appear to be similar to or even better than those of other plant extracts, such as _Emblica officinalis_ or alpha-naphthoflavone, etc. [52; 53]. However, these reports did not show and/or correlate changes in the production of collagen and the inhibition of MMP-1 with the effects on reversing mitochondrial dysfunction. Compared to those reports, the results of the present study clearly showed that the AF increased collagen production by restoring mitochondrial activities, although the mitochondria of human skin fibroblast cells were damaged by UVb light. Therefore, these results indicate that the AF must play an important role in preventing the skin aging process by effectively increasing collagen production and inhibiting MMP-1 production in the presence of exogenous aging factors, such as UVb light.