Cellular Anti-Melanogenic Effects of a Euryale ferox Seed Extract Ethyl Acetate Fraction via the Lysosomal Degradation Machinery
Abstract
:1. Introduction
2. Results
2.1. In Vitro Antioxidant Activities of E. ferox Seed Ethyl Acetate Fraction (Efse-EA)
Compound | Concentration (µg/mL) | DPPH (1) | FRAP (2) | CUPRAC (3) | ABTS (4) |
---|---|---|---|---|---|
Activity (% of Control) | |||||
Ascorbic acid | 1 | 4.5 ± 0.011 (5) | 12.7 ± 0.002 ** | 12.2 ± 0.001 | −1.7 ± 0.002 |
3 | 8.5 ± 0.016 **,(6) | 52.5 ± 0.001 ** | 42.7 ± 0.001 ** | 2.7 ± 0.001 ** | |
10 | 18.5 ± 0.054 * | 170.8 ± 0.006 ** | 147.8 ± 0.001 ** | 18.1 ± 0.003 ** | |
30 | 57.3 ± 0.069 ** | 523.3 ± 0.026 ** | 119.2 ± 0.002 ** | 57.9 ± 0.008 ** | |
Efse-EA | 1 | 0.6 ± 0.002 | 19.9 ± 0.003 ** | 12.8 ± 0.003 * | 5.2 ± 0.001 ** |
3 | 7.9 ± 0.012 | 82.0 ± 0.009 ** | 54.3 ± 0.009 ** | 18.6 ± 0.013 * | |
10 | 28.1 ± 0.006 ** | 234.4 ± 0.027 ** | 251.9 ± 0.029 ** | 55.1 ± 0.024 * | |
30 | 78.0 ± 0.021 ** | 820.4 ± 0.065 ** | 669.6 ± 0.043 ** | 89.7 ± 0.001 * |
2.2. Effect of Efse-EA on Mushroom Tyrosinase and Copper Chelation
2.3. Effect of Efse-EA on Cell Viability and Melanin Levels
2.4. Effects of Efse-EA on l-DOPA (l-3,4-Dihydroxyphenylalanine) Zymography and on Cellular Tyrosinase Activity
2.5. Effect of Efse-EA on Proteins Involved in Melanogenesis and Lysosomal Degradation
3. Discussion
4. Experimental Section
4.1. Materials
4.2. Cell Cultures
4.3. Preparation of Plant Extracts
4.4. Cell Viability Assay
4.5. Evaluation of in Vitro Antioxidant Activities
4.6. Measurement of Tyrosinase and Copper Chelating Activity
4.7. Melanogenesis Inhibition Assay in Melan-a Cells
4.8. Analysis of Intracellular Tyrosinase by Zymography
4.9. Reverse-Transcription Polymerase Chain Reaction (RT-PCR) Analysis
4.10. Western Blot Analysis
4.11. β-Hexosaminidase Assay
4.12. Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Baek, S.-H.; Nam, I.-J.; Kwak, H.S.; Kim, K.-C.; Lee, S.-H. Cellular Anti-Melanogenic Effects of a Euryale ferox Seed Extract Ethyl Acetate Fraction via the Lysosomal Degradation Machinery. Int. J. Mol. Sci. 2015, 16, 9217-9235. https://doi.org/10.3390/ijms16059217
Baek S-H, Nam I-J, Kwak HS, Kim K-C, Lee S-H. Cellular Anti-Melanogenic Effects of a Euryale ferox Seed Extract Ethyl Acetate Fraction via the Lysosomal Degradation Machinery. International Journal of Molecular Sciences. 2015; 16(5):9217-9235. https://doi.org/10.3390/ijms16059217
Chicago/Turabian StyleBaek, Seung-Hwa, In-Jeong Nam, Hyeong Seob Kwak, Ki-Chan Kim, and Sang-Han Lee. 2015. "Cellular Anti-Melanogenic Effects of a Euryale ferox Seed Extract Ethyl Acetate Fraction via the Lysosomal Degradation Machinery" International Journal of Molecular Sciences 16, no. 5: 9217-9235. https://doi.org/10.3390/ijms16059217