Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Neuroprotective effects of cultured and fermented wild ginseng extracts on oxidative stress induced by hydrogen peroxide in PC12 cells

발효산삼배양근농축액의 산화방지 효과 및 과산화수소로 유발된 PC12 세포독성 보호효과

  • 최여옥 (바이오생명공학연구소) ;
  • 김유리 ((주)제이아이바이오팜) ;
  • 신승용 ((주)제이아이바이오팜) ;
  • 이재근 ((주)화진바이오코스메틱) ;
  • 김철중 ((주)화진바이오코스메틱) ;
  • 이예지 ((주)화진바이오코스메틱) ;
  • 강병주 ((주)제이아이바이오팜) ;
  • 김관수 ((주)제이아이바이오팜) ;
  • 최지은 ((주)제이아이바이오팜) ;
  • 한범석 (호서대학교 식품제약공학부)
  • Received : 2018.05.02
  • Accepted : 2018.07.25
  • Published : 2018.08.31
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Abstract

Most neurodegenerative diseases are known to be influenced by oxidative stress. We investigated the anti-oxidative activity of the concentrate of fermented wild ginseng root culture (HLJG0701) containing ginsenosides Rg5 and Rk1. HLJG0701 showed effective DPPH and ABTS radical scavenging ability ($IC_{50}$: 16- and 4-fold dilution, respectively) and was inhibited dose-dependently by the $FeSO_4$-induced lipid peroxidation group (8- and 4-fold dilution: 2.3 and 1.5 nM, respectively). In MTT and LDH assays, 8-, 16-, 32- and 64-fold diluted HLJG0701 significantly increased cell viability by 70, 53, 35, and 26%, respectively. LDH released by HLJG0701 was reduced 1.3-fold with 8-fold diluted HLJG0701 compared to the $H_2O_2$-treated control. In addition, the inhibitory effect of HLJG0701 on oxidative stress in PC12 cells was confirmed by DCF-DA analysis (16-, 4-fold diluted HLJG0701: 50 and 68% ROS inhibition, respectively), TBARS (16- and 4-fold diluted HLJG0701: 50.7 and 46.5% inhibition, respectively), GPx (16- and 4-fold diluted HLJG0701: 133.3 and 227.3% release, respectively), and SOD analysis (16- and 4-fold diluted HLJG0701: 118.2 and 218.2% release, respectively). These results suggested that HLJG0701 protects neuronal cells by its anti-oxidative effects and hence can be a potential preventive material against neurodegenerative diseases.

대부분의 신경퇴행성질환은 산화스트레스의 영향을 받는다. 따라서 본 연구에서는 진세노사이드 Rg5와 Rk1을 함유하고 있는 발효산삼배양근 농축액인 HLJG0701의 산화방지 효능을 확인하였다. 그 결과, HLJG0701는 효과적인 DPPH와 ABTS 라디칼제거능을 나타냈으며($IC_{50}$: 16배, 4배 희석액), 황산철(II) 유도군의 지방질 과산화물 발생 비교 시, 용량의존적으로 억제됨을 확인하였다(8배 희석액: 2.3 nM, 4배 희석액: 1.5 nM). HLJG0701의 세포 보호 효과를 MTT, LDH 분석으로 평가한 결과, 8, 16, 32, 64배 희석액의 HLJG0701를 처리한 경우 70, 53, 35, 26%의 세포생존율이 증가됨을 확인하였다. LDH 방출 측정 결과, $H_2O_2$로 인해 3.6배까지 증가한 LDH 활성은 8배 희석액의 HLJG0701 처리 시 1.3배 수준까지 감소하였다. 또한 HLJG0701가 PC12세포에 발생한 산화스트레스 DCF-DA 분석(16배 희석액: 50% ROS 억제, 4배 희석액: 68% ROS 억제), TBARS (16배 희석액: 50.7% 감소, 4배 희석액: 46.5% 감소), GPx (16배 희석액: 133.3% 증가, 4배 희석액: 227.3% 증가), 그리고 SOD (16배 희석액: 118.2% 증가, 4배 희석액: 218.2% 증가) 분석으로 확인하였다. 위 결과는 HLJG0701가 산화방지 효능을 매개로 신경세포를 보호함으로써 신경퇴행성질환을 예방할 수 있는 가능성 있는 후보물질이 될 수 있다는 것을 의미하며, 임상시험 등 다양한 연구가 충분하지 않은 상태이므로 앞으로 지속적인 연구가 필요할 것으로 사료된다.

Keywords

References

  1. Ahn YM, Park HS, Kwon KR. Anti-cancer and anti-oxidant efficacies of wild ginseng and cultivated wild ginseng of Korea and China. J. Pharmacopuncture 10: 5-16 (2007)
  2. Ahn S, Siddiqi MH, Aceituno VC, Simu SY, Zhang J, Perez ZEJ, Yang DC. Ginsenoside Rg5: Rk1 attenuates TNF-${\alpha}/IFN-{\gamma}$-induced production of thymus-and activation-regulated chemokine (TARC/CCL17) and LPS-induced NO production via downregulation of NF-${\kappa}B$/p38 MAPK/STAT1 signaling in human keratinocytes and macrophages. In Vitro Cell. Dev-An. 52: 287-295 (2016) https://doi.org/10.1007/s11626-015-9983-y
  3. Andersen JK. Oxidative stress in neurodegeneration: cause or consequence?. Nat. Med. 10: 18-25 (2004) https://doi.org/10.1038/nrn1434
  4. Barnham KJ, Masters CL, Bush AI. Neurodegenerative diseases and oxidative stress. Nat Rev. Drug Discov. 3: 205-214 (2004) https://doi.org/10.1038/nrd1330
  5. Bidchol AM, Wilfred A, Abhijna P, Harish R. Free radical scavenging activity of aqueous and ethanolic extract of Brassica oleracea L. var. italica. Food Bioprocess Tech. 4: 1137-1143 (2011) https://doi.org/10.1007/s11947-009-0196-9
  6. Butterfield DA, Reed T, Perluigi M, De Marco C, Coccia R, Cini C, Sultana R, Elevated protein-bound levels of the lipid peroxidation product, 4-hydroxy-2-nonenal, in brain from persons with mild cognitive impairment. Neurosci. Lett. 397: 170-173 (2006) https://doi.org/10.1016/j.neulet.2005.12.017
  7. Halliwell B. Reactive oxygen species and the central nervous system. J. Neurochem. 59: 1609-1623 (1992) https://doi.org/10.1111/j.1471-4159.1992.tb10990.x
  8. Han JY, Chung KH, Ryu GH. Comparison of physicochemical properties and release characteristics of extruded tissue cultured mountain ginseng. J. Korean Soc. Food Sci. Nutr. 37: 1018-1024 (2008) https://doi.org/10.3746/jkfn.2008.37.8.1018
  9. Han SH, Kim SJ, Yun YW, Nam SY, Lee HJ, Lee BJ. Protective effects of cultured and fermented ginseng extracts against scopolamine-induced memory loss in a mouse model. Lab. Anim. Res. 34: 37-43 (2018) https://doi.org/10.5625/lar.2018.34.1.37
  10. Heo HJ, Kim DO, Choi SJ, Shin DH, Lee CY. Potent inhibitory effect of flavonoids in Scutellaria baicalensis on amyloid ${\beta}$ protein-induced neurotoxicity. J. Agr. Food Chem. 52: 4128-4132 (2004) https://doi.org/10.1021/jf049953x
  11. Hwang CR, Lee SH, Jang GY, Hwang IG, Kim HY, Woo KS, Jeong HS. Changes in ginsenoside compositions and antioxidant activities of hydroponic-cultured ginseng roots and leaves with heating temperature. J. Ginseng Res. 38: 180-186 (2014) https://doi.org/10.1016/j.jgr.2014.02.002
  12. Jeong HR, Choi GN, Kim JH, Kwak JH, Kim YS, Jeong CH, Heo HJ. Nutritional components and their antioxidative protection of neuronal cells of litchi (Litchi chinensis Sonn.) fruit pericarp. Korean J. Food Sci. Technol. 42: 481-487 (2010a)
  13. Jeong HS, Lim CS, Cha BC, Kwon KR. Component analysis of cultivated ginseng, cultivated wild ginseng, and wild ginseng and the change of ginsenoside components in the process of red ginseng. J. Pharmacopuncture 13: 63-77 (2010b)
  14. Jin Y, Kim JH, Hong HD, Kwon J, Lee EJ, Jang M, Lee SY, Han AR, Nam TG, Hong SK, Huh TL, Kang NJ, Lim TG. Ginsenosides Rg5 and Rk1, the skin-whitening agents in black ginseng. J. Functi. Foods 45: 67-74 (2018) https://doi.org/10.1016/j.jff.2018.03.036
  15. Keum YS, Park KK, Lee JM, Chun KS, Park JH, Lee SK, Kwon H, Surh YJ. Antioxidant and anti-tumor promoting activities of the methanol extract of heat-processed ginseng. Cancer Lett. 150: 41-48 (2000) https://doi.org/10.1016/S0304-3835(99)00369-9
  16. Kim HS, Kang YH. Antioxidant activity of ethanol extracts of nonedible parts (stalk, stem. leaf, seed) from oriental melon. Korean J. Plant Resour. 23: 451-457 (2010)
  17. Kim JW, Lee SH, No Hk, Hong JH, Youn KS. Antioxidant properties of cultured wild ginseng roots extracts. Korean J. Food Preserv. 17: 861-866 (2010)
  18. Kim DJ, Seong KS, Kim DW, Kim SR, Chang CC. Antioxidative effects of red ginseng saponins on paraquat-induced oxidative stress. J. Ginseng Res. 28: 5-10 (2004) https://doi.org/10.5142/JGR.2004.28.1.005
  19. Kobashigawa LC, Xu YC, Padbury JF, Tseng YT, Yano N. Metformin protects cardiomyocyte from doxorubicin induced cytotoxicity through an AMP-activated protein kinase dependent signaling pathway: an in vitro study. PLoS One 9: e104888 (2014) https://doi.org/10.1371/journal.pone.0104888
  20. Lee SE, Lee SW, Bang JK, Yu YJ, Seong NS. Antioxidant activities of leaf stem and root of Panax ginseng CA Meyer. Korean J. Med. Crop Sci. 12: 237-242 (2004)
  21. Lee JG, Lee YY, Kim SY, Pyo JS, Yun-Choi HS, Park JH. Platelet antiaggregating activity of ginsenosides isolated from processed ginseng. Pharmazie 64: 602-604 (2009b)
  22. Lee SM, Shon HJ, Choi CS, Hung TM, Min BS, Bae K. Ginsenosides from heat processed ginseng. Chem. Pharm. Bull. 57: 92-94 (2009a) https://doi.org/10.1248/cpb.57.92
  23. Lim SJ, Kim YS. Suppressed synthesis of reactive oxygen species from HL60 cell by nicotine. Cancer Prev Res. 12: 35-40 (2007)
  24. Maroto R, Perez-Polo JR. BCL-2-related protein expression in apoptosis: oxidative stress versus serum deprivation in PC12 cells. J. Neurochem. 69: 514-523 (1997)
  25. Park SJ, Yoo SM, Kim YE. Nutritional characteristics and screening of biological activity of cultured wild ginseng roots. Korean J. Food Nutr. 25: 729-736 (2012) https://doi.org/10.9799/ksfan.2012.25.4.729
  26. Polidori MC, Mattioli P, Aldred S, Cecchetti R, Stahl W, Griffiths H, Mecocci P. Plasma antioxidant status, immunoglobulin g oxidation and lipid peroxidation in demented patients: relevance to Alzheimer disease and vascular dementia. Dement. Geriatr. Cogn. 18: 265-270 (2004) https://doi.org/10.1159/000080027
  27. Ramesh T, Kim SW, Hwang SY, Sohn SH, Yoo SK, Kim SK. Panax ginseng reduces oxidative stress and restores antioxidant capacity in aged rats. Nutr. Res. 32: 718-726 (2012) https://doi.org/10.1016/j.nutres.2012.08.005
  28. Shutenko Z, Henry Y, Pinard E, Seylaz J, Potier P, Berthet F, Sercombe R. Influence of the antioxidant quercetin in vivo on the level of nitric oxide determined by electron paramagnetic resonance in rat brain during global ischemia and reperfusion. Biochem. Pharmacol. 57: 199-208 (1999) https://doi.org/10.1016/S0006-2952(98)00296-2
  29. Simu SY, Ahn S, Veronica CA, Yang DC. Ginsenoside Rg5: Rk1 exerts an anti-obesity effect on 3T3-L1 cell line by the downregulation of $PPAR{\gamma}\;and\;CEBP{\alpha}$. Iran. J Biotech. 15: 252-259 (2017) https://doi.org/10.15171/ijb.1517
  30. Yobimoto K, Matsumoto K, Huong NTT, Kasai R, Yamasaki K, Watanabe H. Suppressive effects of Vietnamese ginseng saponin and its major component majonoside-R2 on psychological stressinduced enhancement of lipid peroxidation in the mouse brain. Pharmacol. Biochem. Be. 66: 661-665 (2000) https://doi.org/10.1016/S0091-3057(00)00257-4