The Investigation of the Induction of Diketocarotenoids Senescence in SHSY-5Y Cells

Document Type : Original Article

Authors

• Zahra Zare Dorahi ¹
• Mojtaba Ilani ²
• Mohabbat Jamhiri ³
• Afrooz Daneshparvar ¹
• Shahrokh Zare ¹
• Meysam Zare ⁴
• Iman Jamhiri ¹

¹ Stem Cell Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.

² Department of Physiology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran.

³ Department of Medical Physiology, Faculty of Medicine Shahid Sadoughi University of Medical Sciences, Yazd, Iran

⁴ Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.

Abstract

Introduction: Recently, studies of diketocarotenoids such as astaxanthin (Ax) and canthaxanthin (Cx) with powerful antioxidant have focused on numerous biological mechanisms such as singlet oxygen quenching, radical scavenging, anti-diabetic, anti-carcinogenesis, anti-inflammatory, anti-obesity and anti-melanogenesis activities. There is evidence demonstrating that diketocarotenoid confers neuroprotective effects in experimental models of chronic neurodegenerative disorders and neurological diseases. This study used Ax and Cx to detect its role on senescence of SHSY-5Y Cells. Methods: In this study, the sample included the cell control group (SH-SY5Y cell line) that did not receive Ax and Cx, , and the experimental group that received Ax and Cx (20 mM). Ax and Cx were treated with SH-SY5Y cell line at 48 hours. To measure the expression of BAX, Bcl-2 and PPARγ different groups were compared by real‑time PCR analysis. The cell senescence effects of Ax and Cx, a β-galactosidase (SA-β-gal) senescence assay was evaluated. The results were analyzed by the one-way analysis of variance (ANOVA) using Prism version 6.0 software. Results: The results showed that treatment with Ax and Cx (20 mM) for 48h induced apoptosis and senescence. The BAX and Bcl-2 gene expression analysis revealed a significant impact of Ax and Cx in apoptosis induction (P<0.05). The measuring of cell senescence also indicated that Ax and Cx exhibited a senescence inductive activity as determined by an increase in β-galactosidase activity and PPARγ gene expression (P<0.05). Conclusion: It appears that Ax and Cx have therapeutic properties in SH-SY5Y cells and can cause the proliferation of these cells to cease. The results suggest that Ax and Cx treatment may be beneficial for therapy of neuroblastoma and neurodegenerative disorders.

Keywords

• Astaxanthin
• Canthaxanthin
• Apoptosis
• Senescence
• Neuroblastoma
• SHSY-5Y Cells

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References

1.   Arbo B, Hoppe J, Rodrigues K, Garcia-Segura L, Salbego C, Ribeiro M. 4′-Chlorodiazepam is neuroprotective against amyloid-beta in organotypic hippocampal cultures. J Steroid Biochem Mol Biol. 2017; 171: 281- 287.

2.   Lee D-H, Kim C-S, Lee YJ. Astaxanthin protects against MPTP/MPP+-induced mitochondrial dysfunction and ROS production in vivo and in vitro. Food Chem Toxicol. 2011; 49: 271- 280.

3.   Brizio P, Benedetto A, Righetti M, Prearo M, Gasco L, et al. Astaxanthin and canthaxanthin (xanthophyll) as supplements in rainbow trout diet: in vivo assessment of residual levels and contributions to human health. J Agric Food Chem. 2013; 61: 10954- 10959

4.   Ribeiro D, Freitas M, Silva AM, Carvalho F, Fernandes E. Antioxidant and pro-oxidant activities of carotenoids and their oxidation products. Food Chem Toxicol. 2018; 120: 681- 699.

5.   Widomska J, Subczynski WK. Mechanisms enhancing the protective functions of macular xanthophylls in the retina during oxidative stress. Exp Eye Res. 2018; 4835 (18): 30210- 30220.

6.   Satomi Y. Antitumor and cancer-preventative function of fucoxanthin: a marine carotenoid. Anticancer Res. 2017; 37 (4): 1557- 1562.

7.   Guerin M, Huntley ME, Olaizola M. Haematococcus astaxanthin: applications for human health and nutrition. Trends Biotechnol. 2003; 21 (5): 210- 216.

8.   Chew BP, Park JS. Carotenoid action on the immune response. J Nutr. 2004; 134 (1): 257S- 261S.

9.   Shin J, Kim J-E, Pak K-J, Kang JI, Kim T-S, et al. A Combination of soybean and haematococcus extract alleviates ultraviolet B- induced photoaging. Int J Mol Sci. 2017; 18 (3): 1- 8.

10. Debacq-Chainiaux F, Erusalimsky JD, Campisi J, Toussaint O. Protocols to detect senescence-associated beta-galactosidase (SA-βgal) activity, a biomarker of senescent cells in culture and in vivo. Nat Protoc. 2009; 4 (12): 1798- 1806.

11. Tung Y-C, Tsai M-L, Kuo F-L, Lai C-S, Badmaev V, et al. Se-methyl-L-selenocysteine induces apoptosis via endoplasmic reticulum stress and the death receptor pathway in human colon adenocarcinoma COLO 205 cells. J Agric Food Chem. 2015; 63 (20): 5008- 5016.

12. Liu X, Yamada N, Osawa T. Assessing the neuroprotective effect of antioxidant food factors by application of lipid-derived dopamine modification adducts. Methods Mol Biol. 2010; 594: 263- 273.

13. Ikeda Y, Tsuji S, Satoh A, Ishikura M, Shirasawa T, Shimizu T. Protective effects of astaxanthin on 6‐hydroxydopamine‐induced apoptosis in human neuroblastoma SH‐SY5Y cells. J Neurochem. 2008; 107 (6): 1730- 1740.

14. Sowmya PR-R, Arathi BP, Vijay K, Baskaran V, Lakshminarayana R. Astaxanthin from shrimp efficiently modulates oxidative stress and allied cell death progression in MCF-7 cells treated synergistically with β-carotene and lutein from greens. Food Chem Toxicol. 2017; 106 (Pt A): 58- 69.

15. Douraghi-Zadeh D, Matharu B, Razvi A, Austen B. The protective effects of the nutraceutical, colostrinin, against Alzheimer’s disease, is mediated via prevention of apoptosis in human neurones induced by aggregated β-amyloid. J Nutr Health Aging. 2009; 13 (6): 522- 527.

16. Liao K-S, Wei C-L, Chen J-C, Zheng H-Y, Chen W-C, et al. Astaxanthin enhances pemetrexed-induced cytotoxicity by downregulation of thymidylate synthase expression in human lung cancer cells. Regul Toxicol Pharmacol. 2016; 81: 353- 361.

17. Wu C, Zhang J, Liu T, Jiao G, Li C, Hu B. Astaxanthin inhibits proliferation and promotes apoptosis of A549 lung cancer cells via blocking JAK1/STAT3 pathway. Chinese J Cell Mol Immun. 2016; 32: 784- 788.

18. Namsi A, Nury T, Hamdouni H, Yammine A, Vejux A, et al. Induction of neuronal differentiation of murine N2a cells by two polyphenols present in the mediterranean diet mimicking neurotrophins activities: resveratrol and apigenin. Diseases J. 2018; 6 (3): pii: E67.

19. Rittié L, Fisher GJ. UV-light-induced signal cascades and skin aging. Ageing Res Rev. 2002; 1 (4): 705- 720.

20. Redfern C, Lovat P, Malcolm A, Pearson A. Gene expression and neuroblastoma cell differentiation in response to retinoic acid: differential effects of 9-cis and all-trans retinoic acid. Eur J Cancer. 1995; 31A (4): 486- 494.

21. Chu PW, Cheung WM, Kwong YL. Differential effects of 9-cis, 13-cis and all-trans retinoic acids on the neuronal differentiation of human neuroblastoma cells. Neuroreport. 2003; 14 (15): 1935- 1939.

22. Fujimura M, Usuki F. Methylmercury induces oxidative stress and subsequent neural hyperactivity leading to cell death through the p38 MAPK-CREB pathway in differentiated SH-SY5Y cells. Neurotoxicology. 2018; 67: 226- 233.

23. Chen J, Kelly PT. Retinoic acid stimulates α-CAMKII gene expression in PC12 cells at a distinct transcription initiation site. J Neurosci. 1996; 16 (18): 5704- 5714.

24. Shioda N, Fukunaga K. Physiological and pathological roles of CaMKII-PP1 signaling in the brain. Int J Mol Sci. 2017; 19 (1): pii: E20.