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Anti-aging and Anti-oxidation – Salmon Sperm as a Substitute for Nucleotide Sources

Chen-Meng Kuan , Kai-Wen Kan, Jia-Haur Chen, Young-Hao Lin, Yung-Hsiang Lin
Journal of Food and Nutrition Research. 2019, 7(9), 652-655. DOI: 10.12691/jfnr-7-9-5
Received August 10, 2019; Revised September 14, 2019; Accepted September 20, 2019

Abstract

This research unveils the possibility of a salmon sperm formula, called DNA drink, for anti-aging and anti-oxidation. The applications of salmon sperm have been rarely reported in scientific investigations if compared with salmon roe, but salmon sperm also contains copious nucleotides. Our experiments confirmed that the DNA drink could enhance the anti-oxidant ability and improve the expression levels of aging-related genes (CCT2, CCT6A, Atg1, Atg8, and SIRT1 genes) in peripheral blood mononuclear cells (PBMCs). These results suggest the potential of salmon sperm for boosting cell vitality and protecting cells from oxidative damage.

1. Introduction

Human ambition for longevity continues to advance the development of modern technologies, and human life expectancy will extend with technological progress. The American population at ages 65 and older will rise from 16% in 2018 (52 million people) to 23% by 2060 (95 million people) 1. Aging is the consequence of physiopathological and progressive deterioration with the passage of time and the damages from chronic oxidative stress will hasten the aging process due, in part, to the impact on the regulatory systems (e.g., endocrine and immune systems) 2. Oxidative stress is associated with the internal and external stressors (e.g., mitochondrial leak, UV radiation, cigarette smoking, etc.) 3. Oxidative stress results from the imbalance of the formation and neutralization of reactive oxygen and nitrogen species (RONS), which leads to the accumulation of oxidative damage to macromolecules (i.e., lipid, protein, and DNA) 4, 5. Reactive oxygen species (ROS) include superoxide anion (O2∙−) and hydroxyl radical (OH), which is converted from hydrogen peroxide (H2O2) through Fenton or Haber–Weiss reaction 2, 6. Superoxide anion may further interact with nitric oxide (NO), and reactive nitrogen species (RNS) resulting 7. Nevertheless, human body possesses with sophisticate antioxidant systems in mitochondria for eliminating oxidative stress though several detoxifying enzymes, such as glutathione peroxidase [GPx]/glutathione reductase [GRx], superoxide dismutases (SOD), etc. 8. Exposing skin to UV radiation, UV-induced ROS may initiate skin-aging-related cascades and elicit metalloproteinase (MMP)-1-mediated aging as well as NF-κB-TNF-α-mediated and inflammation-induced aging 9. Moreover, ROS levels are critically connected to oxidative-induced inflammation and the occurrence/progression of various diseases (e.g., cardiovascular diseases, diabetes, neurodegenerative diseases, or cancers) 10, 11, 12, 13. Nrf2-Keap1 pathway is the main protective system against oxidative stress and electrophiles, but abnormal Nrf2 regulation may facilitate the progression of inflammation and, in turn, cancer formation 14, 15. Accordingly, certain ROS expression is correlated with aging, inflammatory response, and the occurrence of chronic diseases. Dietary nucleotide supplements are indentified as an effective means to alleviate the inflammatory process, oxidative damage, carcinogenic activity, and aging process, along with restoring energy from fatigue 16, 17, 18. Nucleotides involve rudimentary biological functions regarding encoding and deciphering genetic information, modulating metabolism and cell signaling, and as cofactors within enzyme reactions 19. Based on the results of animal studies, dietary nucleotide supplements have been proved to prolong lifespan in tumor-bearing rats by means of long-term nucleotide feeding 20. Apart from pure nucleotides, fish roe (egg) may be an alternative route to nucleotide sources 21, 22. A study reported that salmon roe could enhance the type I collagen production over 125% and inhibit antioxidant genes (e.g., OXR1, TXNRD1, and PRDX family genes) in human fibroblasts 22. These evidences imply that dietary nucleotide supplements have the potential to ameliorate the skin conditions and delay the aging process 23. To the best of our knowledge, few research groups unveil the salmon sperm as a nucleotide alternative substitution with respect to the utility in anti-aging or anti-inflammation. In this research, we try to investigate such possibilities through using a salmon sperm formula, DNA drink, to evaluate the expression levels of ROS and aging-related genes in PBMCs. We discovered that DNA drink could substantially enhance these gene expression levels and lower ROS levels, which reveals the hope to extend cell lifespan.

2. Material and Methods

2.1. Materials

DNA drink [MelaGene+ Drink, Melaleuca (China); ingredients: salmon sperm extract, yeast powder, brown rice fermented powder, algal-docosahexaenoic acid, snow fungus extract, citric acid monohydrate, pectin, pear juice, fructose, water, plum/apple/vegetable flavors, and potassium sorbate], PBMC (ATCC® PCS-800-011™), growth media [X-VIVOTM 10 (Lonza), 10% fetal bovine serum, 1 mM sodium pyruvate, and 1% penicillin/streptomycin], 2′,7′-dichlorofluorescin diacetate [DCFH-DA (Sigma-Aldrich)], phosphate buffered saline [PBS (Gibco)], RNA extraction kit (Genaid Biotech), nCounter® platform (NanoString Technologies)

2.2. ROS Assay

We dispersed 2 × 105 PBMCs in 2 mL of the growth media into each well in 6-well plates. Following 24 hours incubation, we replaced the media with the refresh media, the media with 0.5 mM H2O2, the media with 0.5 mM H2O2, or 0.5%/0.25% DNA drink for the corresponding testing cells, and incubated the cells for an hour. Afterwards, we removed the solutions and washed the cells with PBS solutions twice. And we added 10 μg/mL of DCFH-DA (a ROS indicator) to each well and waited for 40 minutes for the staining reaction. Finally, we collected the cells from wells by use of trypsin and loaded the suspending cells into a cytometry (excitation wavelengths: 450-490 nm; emission wavelengths: 510-550 nm).

2.3. Analysis of mRNA Expression

1.5 × 105 PBMCs in 2 mL of the media with 0.25% of DNA drink were dispersed into each well of 6-well plates. Following 24 hour incubation, we collected the PBMCs and extracted their total RNA by the RNA extraction kit. 75 ng/μL RNA extracts were used as templates for analysis of mRNA expression level through the nCounter® platform. The operation was following the nCounter protocol.

2.4. Statistical Analysis

The statistical analysis for the experimental results was based on t-test; p < 0.05 represented significant difference.

3. Results and Discussion

3.1. Evaluation of ROS Production in PBMC

To evaluate the efficacy of the DNA drink for ant-oxidation, following the treatments of 0.5% and 0.25% of DNA drink, we treated PBMCs with 0.5 mM of hydrogen peroxide (H2O2) for an hour to induce the formation and accumulation of ROS (Figure 1). The ROS levels of the cells treated with DNA drink were at least 48% lower than that of the only H2O2-induced cells. We used 0.25% of the salmon sperm formula for the following testing considering sample saving and the similar result with 0.5%.

3.2. mRNA Expression Level of SOD2 Gene

Figure 2 shows the salmon sperm formula could increase the expression level of SOD2 gene, which encodes superoxide dismutase 2, by ~30%. This result corresponds with the ROS elimination result (Figure 1) and verifies the antioxidant capacity in salmon sperm.

3.3. mRNA Expression Levels of Anti-aging Related Genes

Furthermore, we analyzed the mRNA expression levels for some aging-related genes. Figure 3 shows the increase of the expression levels of CCT2, CCT6A, Atg1, Atg8, and SIRT1 genes after DNA drink treatment; especially, the significant changes in the expression of CCT2, CCT6A, and Atg genes. Chaperonin containing TCP1 (CCP) family chaperonins participate in correct protein folding and their high expression levels represent vigorous cell proliferation 24. Autophagy-related proteins (Atg) are associated with the regulation of autophagic process to remove damaged and unnecessary molecules or components in cells by lysosomal degradation, which remains the stable cell functions, structures, and viability 25. Sirtuin 1 (SIRT 1) can deacetylate various cellular proteins and is essential to the control of cell aging, oxidative stress, inflammatory responses, etc. 26. Therefore, salmon sperm indeed can improve cell vitality and the functionalities of the depletion of cell metabolites as well as prevent cells from oxidative damage.

4. Conclusion

This work successfully demonstrates the potential of our salmon sperm formula for anti-aging and anti-oxidation. DNA drink can remarkably improve the anti-oxidative capability of ROS elimination and the increment of the expression level of SOD2 gene. More importantly, DNA drink is able to up-regulate the aging-related genes, which contribute to correct protein folding, the depletion of unnecessary molecules or substances, and the modulation of cell proliferation/aging. In summary, we have proved that salmon sperm can be an alternative route to nucleotide sources apart from salmon roe and improve cell vitality by comprehensive gene regulations.

Acknowledgements

We would like to thanks TCI and TCI GENE research groups for the technical support and Melaleuca (China) for experimental material support.

References

[1]  Kinney, M., Seider, J., Beaty, A.F., Coughlin, K., Dyal, M., Clewley, D., The impact of therapeutic alliance in physical therapy for chronic musculoskeletal pain: a systematic review of the literature. Physiotherapy Theory and Practice, 2018. 28: p. 1-13.
In article      View Article  PubMed
 
[2]  Liguori, I., et al., Oxidative stress, aging, and diseases. Clinical Interventions in Aging, 2018. 13: p. 757-772.
In article      View Article  PubMed  PubMed
 
[3]  Wu, B.-H., Wang, W.-C. and Kuo, H.-Y., Effect of multi-berries drink on endogenous antioxidant activity in subjects who are regular smokers or drinkers. Journal of Food and Nutrition Research, 2016. 4: p. 289-295.
In article      
 
[4]  Beckman, K.B. and Ames, B.N., The free radical theory of aging matures. Physiological Review, 1998. 78: p. 547-581.
In article      View Article  PubMed
 
[5]  Davalli, P., Mitic, T., Caporali, A., Lauriola, A. and D'Arca, D., ROS, cell senescence, and novel molecular mechanisms in aging and age-related diseases. Oxidative Medicine and Cellular Longevity, 2016. 2016: p. 3565127.
In article      View Article  PubMed  PubMed
 
[6]  Das, T.K., Wati, M.R., Fatima-Shad, K., Oxidative stress gated by Fenton and Haber Weiss reactions and its association with Alzheimer’s disease. Archives of Neuroscience, 2015. 2: p. e20078.
In article      View Article
 
[7]  Panth, N., Paudel, K.R., Parajuli, K., Reactive oxygen species: a jey hallmark of cardiovascular disease. Advances in Medicine, 2016. 2016: p. 9152732.
In article      View Article  PubMed  PubMed
 
[8]  Gil Del Valle, L., Oxidative stress in aging: theoretical outcomes and clinical evidences in humans. Biomedicine & Aging Pathology, 2011. 1: p. 1-7.
In article      View Article
 
[9]  Kageyama, H. and Waditee-Sirisattha, R., Antioxidative, anti-inflammatory, and anti-aging properties of mycosporine-like amino acids: molecular and cellular mechanisms in the protection of skin-aging. Marine Drugs, 2019. 17: p. 222.
In article      View Article  PubMed  PubMed
 
[10]  Fakhruddin, S., Alanazi, W. and Jackson, K.E., Diabetes-induced reactive oxygen species: mechanism of their generation and role in renal injury. Journal of Diabetes Research, 2017. 2017: p. 8379327.
In article      View Article  PubMed  PubMed
 
[11]  El-Kenawi, A. and Ruffell, B., Inflammation, ROS, and mutagenesis. Cancer Cell, 2017. 32, p. 727-729.
In article      View Article  PubMed
 
[12]  Kim, G.H., Kim, J.E., Rhie, S.J., Yoon, S., The role of oxidative stress in neurodegenerative diseases. Experimental Neurobiology, 2015. 24: p. 325-340.
In article      View Article  PubMed  PubMed
 
[13]  Adams, L., Franco, M.C., Estevez, A.G., Reactive nitrogen species in cellular signaling. Experimental Biology and Medicine, 2015. 240: p. 711-717.
In article      View Article  PubMed  PubMed
 
[14]  Tu, W., Wang, H., Li, S., Liu, Q., Sha, H., The anti-inflammatory and anti-oxidant mechanisms of the Keap1/Nrf2/ARE signaling pathway in chronic diseases. Aging and Disease, 2019. 10: 637-651.
In article      View Article  PubMed  PubMed
 
[15]  Kansanen, E., Kuosmanen, S.M., Leinonen, H., Levonen, A.L., The Keap1-Nrf2 pathway: mechanisms of activation and dysregulation in cancer. Redox Biology, 2013. 1: p. 45-49.
In article      View Article  PubMed  PubMed
 
[16]  Xu, M., Liang, R., Li, Y., Wang, J., Anti-fatigue effects of dietary nucleotides in mice. Food & Nutrition Research, 2017. 61: p. 1334485.
In article      View Article  PubMed  PubMed
 
[17]  Xu, M., Zhao, M., Yang, R., Zhang, Z., Li, Y., Wang, J., Effect of dietary nucleotides on immune function in Balb/C mice. International Immunopharmacology, 2013. 17: p. 50-56.
In article      View Article  PubMed
 
[18]  Salobir, J., Rezar, V., Pajk, T., Levart, A., Effect of nucleotide supplementation on lymphocyte DNA damage induced by dietary oxidative stress in pigs. Animal Science, 2005. 81: p. 135-140.
In article      View Article
 
[19]  Shiau, S.Y., Gabaudan, J., Lin, Y.H., Dietary nucleotide supplementation enhances immune responses and survival to Streptococcus iniae in hybrid tilapia fed diet containing low fish meal. Aquaculture Reports, 2015. 2: p. 77-81.
In article      View Article
 
[20]  Xu, M., et al., Dietary nucleotides extend the life span in Sprague-Dawley rats. The Journal of Nutrition, Health & Aging, 2013. 17: p. 223-229.
In article      View Article  PubMed
 
[21]  Marotta, F., et al., Beneficial modulation from a high-purity caviar-derived homogenate on chronological skin aging. Rejuvenation Research, 2012. 15: p. 174-177.
In article      View Article  PubMed
 
[22]  Yoshino, A., Polouliakh, N., Meguro, A., Takeuchi, M., Kawagoe, T. and Mizuki, N., Chum salmon egg extracts induce upregulation of collagen type I and exert antioxidative effects on human dermal fibroblast cultures. Clinical Interventions in Aging, 2016. 11: p. 1159-1168.
In article      View Article  PubMed  PubMed
 
[23]  Zhang, S. and Duan, E., Fighting against skin aging: the way from Bench to Bedside. Cell Transplantation, 2018. 27: p. 729-738.
In article      View Article  PubMed  PubMed
 
[24]  Noormohammadi, A., Somatic increase of CCT8 mimics proteostasis of human pluripotent stem cells and extends C. elegans lifespan. Nature Communication, 2016. 7: p. 13649.
In article      View Article  PubMed  PubMed
 
[25]  Lee, Y.-K., Lee, J.-A., Role of the mammalian ATG8/LC3 family in autophagy: differential and compensatory roles in the spatiotemporal regulation of autophagy. BMB Reports, 2016. 49: p. 424-430.
In article      View Article  PubMed  PubMed
 
[26]  Salminen, A., Kaarniranta, K., Kauppinen, A., Crosstalk between oxidative stress and SIRT1: impact on the aging process. International Journal of Molecular Sciences, 2013. 14: p. 3834-3859.
In article      View Article  PubMed  PubMed
 

Published with license by Science and Education Publishing, Copyright © 2019 Chen-Meng Kuan, Kai-Wen Kan, Jia-Haur Chen, Young-Hao Lin and Yung-Hsiang Lin

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

Cite this article:

Normal Style
Chen-Meng Kuan, Kai-Wen Kan, Jia-Haur Chen, Young-Hao Lin, Yung-Hsiang Lin. Anti-aging and Anti-oxidation – Salmon Sperm as a Substitute for Nucleotide Sources. Journal of Food and Nutrition Research. Vol. 7, No. 9, 2019, pp 652-655. http://pubs.sciepub.com/jfnr/7/9/5
MLA Style
Kuan, Chen-Meng, et al. "Anti-aging and Anti-oxidation – Salmon Sperm as a Substitute for Nucleotide Sources." Journal of Food and Nutrition Research 7.9 (2019): 652-655.
APA Style
Kuan, C. , Kan, K. , Chen, J. , Lin, Y. , & Lin, Y. (2019). Anti-aging and Anti-oxidation – Salmon Sperm as a Substitute for Nucleotide Sources. Journal of Food and Nutrition Research, 7(9), 652-655.
Chicago Style
Kuan, Chen-Meng, Kai-Wen Kan, Jia-Haur Chen, Young-Hao Lin, and Yung-Hsiang Lin. "Anti-aging and Anti-oxidation – Salmon Sperm as a Substitute for Nucleotide Sources." Journal of Food and Nutrition Research 7, no. 9 (2019): 652-655.
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  • Figure 1. Evaluation of ROS production in PBMCs under the circumstance of different concentrations of DNA drink. (n = 3, mean value ± S.D.) (Comparison with the control group: ###, p < 0.001) (Comparison with the H2O2 group: ***, p < 0.001; **, p < 0.01)
  • Figure 2. The mRNA expression levels of SOD2 gene in PBMCs after 0.25% DNA drink treatment. (n = 3, mean value ± S.D.; **, p < 0.01)
  • Figure 3. The mRNA expression levels of anti-aging related genes after 0.25% DNA drink treatment. (n = 3, mean value ± S.D.; ***, p < 0.001)
[1]  Kinney, M., Seider, J., Beaty, A.F., Coughlin, K., Dyal, M., Clewley, D., The impact of therapeutic alliance in physical therapy for chronic musculoskeletal pain: a systematic review of the literature. Physiotherapy Theory and Practice, 2018. 28: p. 1-13.
In article      View Article  PubMed
 
[2]  Liguori, I., et al., Oxidative stress, aging, and diseases. Clinical Interventions in Aging, 2018. 13: p. 757-772.
In article      View Article  PubMed  PubMed
 
[3]  Wu, B.-H., Wang, W.-C. and Kuo, H.-Y., Effect of multi-berries drink on endogenous antioxidant activity in subjects who are regular smokers or drinkers. Journal of Food and Nutrition Research, 2016. 4: p. 289-295.
In article      
 
[4]  Beckman, K.B. and Ames, B.N., The free radical theory of aging matures. Physiological Review, 1998. 78: p. 547-581.
In article      View Article  PubMed
 
[5]  Davalli, P., Mitic, T., Caporali, A., Lauriola, A. and D'Arca, D., ROS, cell senescence, and novel molecular mechanisms in aging and age-related diseases. Oxidative Medicine and Cellular Longevity, 2016. 2016: p. 3565127.
In article      View Article  PubMed  PubMed
 
[6]  Das, T.K., Wati, M.R., Fatima-Shad, K., Oxidative stress gated by Fenton and Haber Weiss reactions and its association with Alzheimer’s disease. Archives of Neuroscience, 2015. 2: p. e20078.
In article      View Article
 
[7]  Panth, N., Paudel, K.R., Parajuli, K., Reactive oxygen species: a jey hallmark of cardiovascular disease. Advances in Medicine, 2016. 2016: p. 9152732.
In article      View Article  PubMed  PubMed
 
[8]  Gil Del Valle, L., Oxidative stress in aging: theoretical outcomes and clinical evidences in humans. Biomedicine & Aging Pathology, 2011. 1: p. 1-7.
In article      View Article
 
[9]  Kageyama, H. and Waditee-Sirisattha, R., Antioxidative, anti-inflammatory, and anti-aging properties of mycosporine-like amino acids: molecular and cellular mechanisms in the protection of skin-aging. Marine Drugs, 2019. 17: p. 222.
In article      View Article  PubMed  PubMed
 
[10]  Fakhruddin, S., Alanazi, W. and Jackson, K.E., Diabetes-induced reactive oxygen species: mechanism of their generation and role in renal injury. Journal of Diabetes Research, 2017. 2017: p. 8379327.
In article      View Article  PubMed  PubMed
 
[11]  El-Kenawi, A. and Ruffell, B., Inflammation, ROS, and mutagenesis. Cancer Cell, 2017. 32, p. 727-729.
In article      View Article  PubMed
 
[12]  Kim, G.H., Kim, J.E., Rhie, S.J., Yoon, S., The role of oxidative stress in neurodegenerative diseases. Experimental Neurobiology, 2015. 24: p. 325-340.
In article      View Article  PubMed  PubMed
 
[13]  Adams, L., Franco, M.C., Estevez, A.G., Reactive nitrogen species in cellular signaling. Experimental Biology and Medicine, 2015. 240: p. 711-717.
In article      View Article  PubMed  PubMed
 
[14]  Tu, W., Wang, H., Li, S., Liu, Q., Sha, H., The anti-inflammatory and anti-oxidant mechanisms of the Keap1/Nrf2/ARE signaling pathway in chronic diseases. Aging and Disease, 2019. 10: 637-651.
In article      View Article  PubMed  PubMed
 
[15]  Kansanen, E., Kuosmanen, S.M., Leinonen, H., Levonen, A.L., The Keap1-Nrf2 pathway: mechanisms of activation and dysregulation in cancer. Redox Biology, 2013. 1: p. 45-49.
In article      View Article  PubMed  PubMed
 
[16]  Xu, M., Liang, R., Li, Y., Wang, J., Anti-fatigue effects of dietary nucleotides in mice. Food & Nutrition Research, 2017. 61: p. 1334485.
In article      View Article  PubMed  PubMed
 
[17]  Xu, M., Zhao, M., Yang, R., Zhang, Z., Li, Y., Wang, J., Effect of dietary nucleotides on immune function in Balb/C mice. International Immunopharmacology, 2013. 17: p. 50-56.
In article      View Article  PubMed
 
[18]  Salobir, J., Rezar, V., Pajk, T., Levart, A., Effect of nucleotide supplementation on lymphocyte DNA damage induced by dietary oxidative stress in pigs. Animal Science, 2005. 81: p. 135-140.
In article      View Article
 
[19]  Shiau, S.Y., Gabaudan, J., Lin, Y.H., Dietary nucleotide supplementation enhances immune responses and survival to Streptococcus iniae in hybrid tilapia fed diet containing low fish meal. Aquaculture Reports, 2015. 2: p. 77-81.
In article      View Article
 
[20]  Xu, M., et al., Dietary nucleotides extend the life span in Sprague-Dawley rats. The Journal of Nutrition, Health & Aging, 2013. 17: p. 223-229.
In article      View Article  PubMed
 
[21]  Marotta, F., et al., Beneficial modulation from a high-purity caviar-derived homogenate on chronological skin aging. Rejuvenation Research, 2012. 15: p. 174-177.
In article      View Article  PubMed
 
[22]  Yoshino, A., Polouliakh, N., Meguro, A., Takeuchi, M., Kawagoe, T. and Mizuki, N., Chum salmon egg extracts induce upregulation of collagen type I and exert antioxidative effects on human dermal fibroblast cultures. Clinical Interventions in Aging, 2016. 11: p. 1159-1168.
In article      View Article  PubMed  PubMed
 
[23]  Zhang, S. and Duan, E., Fighting against skin aging: the way from Bench to Bedside. Cell Transplantation, 2018. 27: p. 729-738.
In article      View Article  PubMed  PubMed
 
[24]  Noormohammadi, A., Somatic increase of CCT8 mimics proteostasis of human pluripotent stem cells and extends C. elegans lifespan. Nature Communication, 2016. 7: p. 13649.
In article      View Article  PubMed  PubMed
 
[25]  Lee, Y.-K., Lee, J.-A., Role of the mammalian ATG8/LC3 family in autophagy: differential and compensatory roles in the spatiotemporal regulation of autophagy. BMB Reports, 2016. 49: p. 424-430.
In article      View Article  PubMed  PubMed
 
[26]  Salminen, A., Kaarniranta, K., Kauppinen, A., Crosstalk between oxidative stress and SIRT1: impact on the aging process. International Journal of Molecular Sciences, 2013. 14: p. 3834-3859.
In article      View Article  PubMed  PubMed