Journal of Nutrition and Health

The Korean Nutrition Society (한국영양학회)
권호 표지
DOI QR코드

DOI QR Code

Neuroprotective effect of fermented ginger extracts by Bacillus subtilis in SH-SY5Y cells

고초균에 의한 생강 발효 추출물의 신경세포 보호 효과

  • Yang, Hee Sun (Functional Food & Nutrition Division, Department of Agrofood Resources, National Institute of Agricultural Science, Rural Development Administration) ;
  • Kim, Mi Jin (Functional Food & Nutrition Division, Department of Agrofood Resources, National Institute of Agricultural Science, Rural Development Administration) ;
  • Kim, Mina (Functional Food & Nutrition Division, Department of Agrofood Resources, National Institute of Agricultural Science, Rural Development Administration) ;
  • Choe, Jeong-sook (Functional Food & Nutrition Division, Department of Agrofood Resources, National Institute of Agricultural Science, Rural Development Administration)
  • 양희선 (농촌진흥청 국립농업과학원 농식품자원부) ;
  • 김미진 (농촌진흥청 국립농업과학원 농식품자원부) ;
  • 김민아 (농촌진흥청 국립농업과학원 농식품자원부) ;
  • 최정숙 (농촌진흥청 국립농업과학원 농식품자원부)
  • Received : 2021.11.15
  • Accepted : 2021.12.16
  • Published : 2021.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

Purpose: The ginger rhizome (Zingiber officinale) is widely cultivated as a spice for its aromatic and pungent components. One of its constituents, 6-hydroxydopamine (6-OHDA) is usually thought to cross the cell membrane through dopamine uptake transporters, and induce inhibition of mitochondrial respiration and the generation of intracellular reactive oxygen species (ROS). This study examines the neuroprotective effect and acetylcholinesterase (AChE) inhibitory activity of fermented ginger extracts (FGEs) on 6-OHDA induced toxicity in SH-SY5Y human neuroblastoma cells. Methods: Ginger was fermented using 2 species of Bacillus subtilis, with or without enzyme pretreatment. Each sample was extracted with 70% ethanol. Neurotoxicity was assessed by applying the EZ-Cytox cell viability assay and by measuring lactic dehydrogenase (LDH) release. Morphological changes of apoptotic cell nuclei were observed by Hoechst staining. Cell growth and apoptosis of SH-SY5Y cells were determined by Western blotting and enzyme activity analysis of caspase-3, and AChE enzymatic activity was determined by the colorimetric assay. Results: In terms of cell viability and LDH release, exposure to FGE showed neuroprotective activities against 6-OHDA stimulated stress in SH-SY5Y cells. Furthermore, FGE reduced the 6-OHDA-induced apoptosis, as determined by Hoechst staining. The occurrence of apoptosis in 6-OHDA treated cells was confirmed by determining the caspase-3 activity. Exposure to 6-OHDA resulted in increased caspase-3 activity of SH-SY5Y cells, as compared to the unexposed group. However, pre-treatment with FGE inhibited the activity of caspase-3. The neuroprotective effects of FGE were also found to be caspase-dependent, based on reduction of caspase-3 activity. Exposure to FGE also inhibited the activity of AChE induced by 6-OHDA, in a dose-dependent manner. Conclusion: Taken together, our results show that FGE exhibits a neuroprotective effect in 6-OHDA treated SH-SY5Y cells, thereby making it a potential novel agent for the prevention or treatment of neurodegenerative disease.

본 연구에서는 생강을 효소처리하여 수용화율을 높인 후 고초균을 이용하여 발효시킨 생강발효물 (FGEs)을 제조하고, 이를 SH-SY5Y에 6-OHDA와 함께 처리하여 세포 보호 효과 및 AChE 저해 활성을 평가하였다. 그 결과, 6-OHDA로 자극된 신경세포에서 FGE를 처리한 모든 실험군에서 세포 생존율이 증가하고 LDH 농도가 감소하였다. 6-OHDA로 유발된 세포자멸사를 억제할 수 있는지 확인하기 위해 핵의 형태학적인 변화 및 caspase-3 활성을 확인하였다. FGE를 처리한 모든 실험군에서 핵의 손상 및 apoptotic body의 감소와 caspase-3 억제 활성을 확인할 수 있었다. 또한 FGE는 AChE의 활성을 유의적으로 감소시켰다. 시료 간의 활성 차이를 비교하였을 때, 생강에 효소 처리 후 고초균으로 발효한 추출물군 (E/BKG와 E/BCG)의 신경세포 보호 활성이 효소 처리하지 않은 발효생강군 (BKG와 BCG) 보다 유의적으로 크게 나타났다. 그러나 발효에 사용된 고초균 2종 간의 활성은 유사하였으며 처리군 간 유의적인 차이를 보이지 않았다. 본 연구 결과로부터 효소처리하여 수용화율을 높여 고초균으로 발효한 FGE는 신경세포 보호 및 AChE 저해 효과를 나타내어 향후 신경질환 연구를 위한 기초자료 제공 및 고부가가치 식품소재 개발에 이용 가능할 것으로 판단된다.

Keywords

Acknowledgement

This work was carried out with the support of "Cooperative Research Program for Agriculture Science and Technology Development (project No. PJ01511502)" Rural Development Administration, Republic of Korea.

References

  1. Lee SI, Choi CH, Kim JS, Lim SS, Jung HW. The antioxidant activities and neuroprotective effects of hot water extracts from torreyae semen. Korean J Herbol 2017; 32(6): 41-48. https://doi.org/10.6116/kjh.2017.32.1.41.
  2. Park EK, Han KH, Lee SH, Kim NK, Bae MH, Seo YH, et al. Neuroprotective effects of Schisandra chinensis and Ribes fasciculatum extract on hydrogen peroxide-mediated oxidative stress in neuroblastic SH-SY5Y cell line. Korean J Food Nutr 2018; 31(6): 865-872. https://doi.org/10.9799/KSFAN.2018.31.6.865
  3. Cho H, Kang K. Effects of taro extract on brain resilience in in vitro Parkinson's disease model induced by 6-hydroxydopamine. J Korean Biol Nurs Sci 2020; 22(4): 223-231. https://doi.org/10.7586/jkbns.2020.22.4.223
  4. Martin HL, Teismann P. Glutathione--a review on its role and significance in Parkinson's disease. FASEB J 2009; 23(10): 3263-3272. https://doi.org/10.1096/fj.08-125443
  5. Orr CF, Rowe DB, Halliday GM. An inflammatory review of Parkinson's disease. Prog Neurobiol 2002; 68(5): 325-340. https://doi.org/10.1016/S0301-0082(02)00127-2
  6. Glinka YY, Youdim MB. Inhibition of mitochondrial complexes I and IV by 6-hydroxydopamine. Eur J Pharmacol 1995; 292(3-4): 329-332. https://doi.org/10.1016/0926-6917(95)90040-3
  7. Deumens R, Blokland A, Prickaerts J. Modeling Parkinson's disease in rats: an evaluation of 6-OHDA lesions of the nigrostriatal pathway. Exp Neurol 2002; 175(2): 303-317. https://doi.org/10.1006/exnr.2002.7891
  8. Lee MK, Kang SJ, Poncz M, Song KJ, Park KS. Resveratrol protects SH-SY5Y neuroblastoma cells from apoptosis induced by dopamine. Exp Mol Med 2007; 39(3): 376-384. https://doi.org/10.1038/emm.2007.42
  9. Seo YH. Antioxidant and antimicrobial activities of ginger with aging and fermentation. Korean J Food Preserv 2007; 24(8): 1180-1187. https://doi.org/10.11002/KJFP.2017.24.8.1180
  10. Kim JS, Kor MS, Kim YH, Kim MK, Hong JS. Volatile flavor components of Korean ginger (Zingiber officinale Roscoe). Korean J Food Sci Technol 1991; 23(2): 141-149.
  11. Munda S, Dutta S, Haldar S, Lal M. Chemical analysis and therapeutic uses of ginger (Zingiber officinale Rosc.) essential oil: a review. J Essent Oil Bear Pl 2018; 21(4): 994-1002. https://doi.org/10.1080/0972060x.2018.1524794
  12. Ho SC, Chang KS, Lin CC. Anti-neuroinflammatory capacity of fresh ginger is attributed mainly to 10-gingerol. Food Chem 2013; 141(3): 3183-3191. https://doi.org/10.1016/j.foodchem.2013.06.010
  13. Mashhadi NS, Ghiasvand R, Askari G, Hariri M, Darvishi L, Mofid MR. Anti-oxidative and antiinflammatory effects of ginger in health and physical activity: review of current evidence. Int J Prev Med 2013; 4(Suppl 1): S36-S42.
  14. Ryu HS, Kim HS. Effect of Zingiber officinale Roscoe extracts on mice immune cell activation. Korean J Nutr 2004; 37(1): 23-30.
  15. Kim HS, Choi JH, Lee HJ, Jeong MC, Kim BS, Kim D. Quality characteristics of treated with mild heat and minced ginger during storage. Korean J Food Preserv 2010; 17(6): 784-792.
  16. Chun YG, Chung HY. Quality properties of fermented gingers. Korean J Food Sci Technol 2011; 43(3): 249-254. https://doi.org/10.9721/KJFST.2011.43.3.249
  17. Chung HY, Park SY. Effects of fermented ginger extracts on the production of NO and PGE2 in RAW 264.7 macrophages. Food Eng Prog 2013; 17(1): 55-61.
  18. Nam DG, Kim M, Im P, Kim SB, Choe JS, Choi AJ. Solubilization of polysaccharide and functional components by high-pressure enzyme treatment from ginger (Zingiber officinale Rosc.). Food Eng Prog 2018; 22(2): 173-185. https://doi.org/10.13050/foodengprog.2018.22.2.173
  19. Nam DG, Kim M, Im P, Kim SB, Choe JS, Choi AJ. Solubilization of polysaccharide and functional components of ginger (Zingiber officinale Rosc.) using ethanol and enzyme. Korean J Food Preserv 2019; 26(5): 545-554. https://doi.org/10.11002/kjfp.2019.26.5.545
  20. Ban YJ, Baik MY, Hahm YY, Kim HK, Kim BY. Optimization of processing conditions for making a black ginger and design mixture for black ginger drinks. Food Eng Prog 2011; 14(2): 112-117.
  21. Steckley D, Karajgikar M, Dale LB, Fuerth B, Swan P, Drummond-Main C, et al. Puma is a dominant regulator of oxidative stress induced Bax activation and neuronal apoptosis. J Neurosci 2007; 27(47): 12989-12999. https://doi.org/10.1523/JNEUROSCI.3400-07.2007
  22. Kirkinezos IG, Moraes CT. Reactive oxygen species and mitochondrial diseases. Semin Cell Dev Biol 2001; 12(6): 449-457. https://doi.org/10.1006/scdb.2001.0282
  23. Guo S, Bezard E, Zhao B. Protective effect of green tea polyphenols on the SH-SY5Y cells against 6-OHDA induced apoptosis through ROS-NO pathway. Free Radic Biol Med 2005; 39(5): 682-695. https://doi.org/10.1016/j.freeradbiomed.2005.04.022
  24. 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. https://doi.org/10.1111/j.1471-4159.2008.05743.x
  25. Nagendra chari KL, Manasa D, Srinivas P, Sowbhagya HB. Enzyme-assisted extraction of bioactive compounds from ginger (Zingiber officinale Roscoe). Food Chem 2013; 139(1-4): 509-514. https://doi.org/10.1016/j.foodchem.2013.01.099
  26. Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol 2007; 35(4): 495-516. https://doi.org/10.1080/01926230701320337
  27. Ouyang L, Shi Z, Zhao S, Wang FT, Zhou TT, Liu B, et al. Programmed cell death pathways in cancer: a review of apoptosis, autophagy and programmed necrosis. Cell Prolif 2012; 45(6): 487-498. https://doi.org/10.1111/j.1365-2184.2012.00845.x
  28. Ochu EE, Rothwell NJ, Waters CM. Caspases mediate 6-hydroxydopamine-induced apoptosis but not necrosis in PC12 cells. J Neurochem 1998; 70(6): 2637-2640. https://doi.org/10.1046/j.1471-4159.1998.70062637.x
  29. Schulz V. Ginkgo extract or cholinesterase inhibitors in patients with dementia: what clinical trials and guidelines fail to consider. Phytomedicine 2003; 10 Suppl 4: 74-79. https://doi.org/10.1078/1433-187X-00302
  30. Chung YK, Heo HJ, Kim EK, Kim HK, Huh TL, Lim Y, et al. Inhibitory effect of ursolic acid purified from Origanum majorana L on the acetylcholinesterase. Mol Cells 2001; 11(2): 137-143.
  31. Oh HK, Kim SH. Effect of soy isoflavone intake on water maze performance and brain acetylcholinesterase activity in rats. Korean J Nutr 2006; 39(3): 219-224.
  32. Hansen RA, Gartlehner G, Webb AP, Morgan LC, Moore CG, Jonas DE. Efficacy and safety of donepezil, galantamine, and rivastigmine for the treatment of Alzheimer's disease: a systematic review and metaanalysis. Clin Interv Aging 2008; 3(2): 211-225.