DOI QR코드

DOI QR Code

The beneficial effect of ginsenosides extracted by pulsed electric field against hydrogen peroxide-induced oxidative stress in HEK-293 cells

  • Liu, Di (Laboratory of Nutrition and Functional Food, College of Food Science and Engineering, Jilin University) ;
  • Zhang, Ting (Laboratory of Nutrition and Functional Food, College of Food Science and Engineering, Jilin University) ;
  • Chen, Zhifei (Laboratory of Nutrition and Functional Food, College of Food Science and Engineering, Jilin University) ;
  • Wang, Ying (Laboratory of Nutrition and Functional Food, College of Food Science and Engineering, Jilin University) ;
  • Ma, Shuang (Laboratory of Nutrition and Functional Food, College of Food Science and Engineering, Jilin University) ;
  • Liu, Jiyun (Laboratory of Nutrition and Functional Food, College of Food Science and Engineering, Jilin University) ;
  • Liu, Jingbo (Laboratory of Nutrition and Functional Food, College of Food Science and Engineering, Jilin University)
  • Received : 2015.09.16
  • Accepted : 2016.02.26
  • Published : 2017.04.15

Abstract

Background: Ginsenosides are the main pharmacological components of Panax ginseng root, which are thought to be primarily responsible for the suppressing effect on oxidative stress. Methods: 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and oxygen radical absorption capacity were applied to evaluate the antioxidant activities of the ginsenosides. Human embryonic kidney 293 (HEK-293) cells were incubated with ginsenosides extracted by pulsed electric field (PEF) and solvent cold soak extraction (SCSE) for 24 h and then the injury was induced by $40{\mu}M$ $H_2O_2$. The cell viability and surface morphology of HEK-293 cells were studied using MTS assay and scanning electron microscopy, respectively. Dichloro-dihydro-fluorescein diacetate fluorescent probe assay was used to measure the level of intracellular reactive oxygen species. The intracellular antioxidant activities of ginsenosides were evaluated by cellular antioxidant activity assay in HepG2 cells. Results: The PEF extracts displayed the higher 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and stronger oxygen radical absorption capacity (with an oxygen radical absorption capacity value of $14.48{\pm}4.04{\mu}M\;TE\;per\;{\mu}g/mL$). The HEK-293 cell model also suggested that the protective effect of PEF extracts was dose-dependently greater than SCSE extracts. Dichloro-dihydro-fluorescein diacetate assay further proved that PEF extracts are more active (8% higher than SCSE extracts) in reducing intracellular reactive oxygen species accumulation. In addition, scanning electron microscopy images showed that the HEK-293 cells, which were treated with PEF extracts, maintained more intact surface morphology. Cellular antioxidant activity values indicated that ginsenosides extracted by PEF had stronger cellular antioxidant activity than SCSE ginsenosides extracts. Conclusion: The present study demonstrated the antioxidative effect of ginsenosides extracted by PEF in vitro. Furthermore, rather than SCSE, PEF may be more useful as an alternative extraction technique for the extraction of ginsenosides with enhanced antioxidant activity.

Keywords

References

  1. Pyo CW, Choi JH, Oh SM, Choi SY. Oxidative stress-induced cyclin d1 depletion and its role in cell cycle processing. Biochim Biophys Acta 2013;1830:5316-25. https://doi.org/10.1016/j.bbagen.2013.07.030
  2. Samuni Y, Goldstein S, Dean OM, Berk M. The chemistry and biological activities of N-acetylcysteine. Biochim Biophys Acta 2013;1830:4117-29. https://doi.org/10.1016/j.bbagen.2013.04.016
  3. Zhou W, Chai H, Lin PH, Lumsden AB, Yao QZ, Chen CY. Molecular mechanisms and clinical applications of ginseng root for cardiovascular disease. Med Sci Monit 2004;10:RA187-92.
  4. Lu JM, Weakley SM, Yang Z, Hu M, Yao QZ, Chen CY. Ginsenoside rb1 directly scavenges hydroxyl radical and hypochlorous acid. Curr Pharm Des 2012;18:6339-47. https://doi.org/10.2174/138161212803832254
  5. Ni N, Liu Q, Ren H, Wu D, Luo C, Li P, Wan JB, Su H. Ginsenoside rb1 protects rat neural progenitor cells against oxidative injury. Molecules 2014;19:3012-24. https://doi.org/10.3390/molecules19033012
  6. Jiang Z, Wang Y, Zhang X, Peng T, Li Y, Zhang Y. Protective effect of ginsenoside r0 on anoxic and oxidative damage in vitro. Biomol Ther 2012;20:544-9. https://doi.org/10.4062/biomolther.2012.20.6.544
  7. Ma J, Liu J, Wang Q, Yu H, Chen Y, Xiang L. The beneficial effect of ginsenoside rg1 on Schwann cells subjected to hydrogen peroxide induced oxidative injury. Int J Biol Sci 2013;9:624-36. https://doi.org/10.7150/ijbs.5885
  8. Hou J, He S, Ling M, Li W, Dong R, Pan Y, Zheng Y. A method of extracting ginsenosides from Panax ginseng by pulsed electric field. J Sep Sci 2010;33:2707-13. https://doi.org/10.1002/jssc.201000033
  9. Lim JH, Shim JM, Lee DU, Kim YH, Park K-J. Pulsed electric fields effects on drying of white ginseng and extraction of soluble components. Korean J Food Sci Technol 2012;44:704-10. https://doi.org/10.9721/KJFST.2012.44.6.704
  10. Kang KS, Kim HY, Pyo JS, Yokozawa T. Increase in the free radical scavenging activity of ginseng by heat-processing. Biol Pharm Bull 2006;29:750-4. https://doi.org/10.1248/bpb.29.750
  11. 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 heatprocessed ginseng. Cancer Lett 2000;150:41-8. https://doi.org/10.1016/S0304-3835(99)00369-9
  12. Kim WY, Kim JM, Han SB, Lee SK, Kim ND, Park MK, Kim CK, Park JH. Steaming of ginseng at high temperature enhances biological activity. J Nat Prod 2000;63:1702-4. https://doi.org/10.1021/np990152b
  13. Liu JB, Chen ZF, He J, Zhang Y, Zhang T, Jiang YQ. Anti-oxidative and antiapoptosis effects of egg white peptide, trp-asn-trp-ala-asp, against h2o2-induced oxidative stress in human embryonic kidney 293 cells. Food Funct 2014;5:3179-88. https://doi.org/10.1039/C4FO00665H
  14. Bradshaw E, Beggs G, Cartafalsa M. High glucose-induces oxidative stress in HEK-293 cells. Mol Biol Cell 2014;25:1. https://doi.org/10.1091/mbc.E13-05-0291
  15. Hani SB, Bayachou M. Salvia fruticosa reduces intrinsic cellular and h2o2-induced DNA oxidation in HEK-293 cells; assessment using flow cytometry. Asian Pac J Trop Biomed 2014;4:399-403. https://doi.org/10.12980/APJTB.4.2014C1270
  16. Pu Z, Zhang PJ, Gao YG, Hao JX, Wang YX, Ran L, He Y, Xue L, Ping L, Zhou DW, et al. The evaluation of contents of nine ginsenoside monomers in ginseng hairy roots by high performance liquid chromatography (HPLC). J Med Plants Res 2011;5:5513-6.
  17. Yokozawa T, Chen CP, Dong E, Tanaka T, Nonaka GI, Nishioka I. Study on the inhibitory effect of tannins and flavonoids against the 1,1-diphenyl-2 picrylhydrazyl radical. Biochem Pharmacol 1998;56:213-22. https://doi.org/10.1016/S0006-2952(98)00128-2
  18. Huang DJ, Ou BX, Hampsch-Woodill M, Flanagan JA, Deemer EK. Development and validation of oxygen radical absorbance capacity assay for lipophilic antioxidants using randomly methylated beta-cyclodextrin as the solubility enhancer. J Agric Food Chem 2002;50:1815-21. https://doi.org/10.1021/jf0113732
  19. Cory AH, Owen TC, Barltrop JA, Cory JG. Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture. Cancer Commun 1991;3:207-12. https://doi.org/10.3727/095535491820873191
  20. Xu J, Hao Z, Gou X, Tian W, Jin Y, Cui S, Guo J, Sun Y, Wang Y, Xu Z. Imaging of reactive oxygen species burst from mitochondria using laser scanning confocal microscopy. Microsc Res Tech 2013;76:612-7. https://doi.org/10.1002/jemt.22207
  21. Delle LE, Lanche R, Law JKY, Weil M, Vu XT, Wagner P, Ingebrandt S. Reduced graphene oxide micropatterns as an interface for adherent cells. Phys Status Solidi A 2013;210:975-82. https://doi.org/10.1002/pssa.201200864
  22. Fang C, Liu Y, Lu P, Liang Q, Liu X, Zhao W, Su W. Total saponins of Panax ginseng inhibiting human endothelium cells' damages induced by angiotensin II via at1 receptor. J Ethnopharmacol 2011;138:439-44. https://doi.org/10.1016/j.jep.2011.09.036
  23. Wolfe KL, Liu RH. Cellular antioxidant activity (caa) assay for assessing antioxidants, foods, and dietary supplements. J Agric Food Chem 2007;55:8896-907. https://doi.org/10.1021/jf0715166
  24. Sanchez-Moreno C. Review: Methods used to evaluate the free radical scavenging activity in foods and biological systems. Food Sci Technol Int 2002;8:121-37. https://doi.org/10.1177/1082013202008003770
  25. Chen R, Meng F, Zhang S, Liu Z. Effects of ultrahigh pressure extraction conditions on yields and antioxidant activity of ginsenoside from ginseng. Sep Purifi Technol 2009;66:340-6. https://doi.org/10.1016/j.seppur.2008.12.026
  26. Huang DJ, Ou BX, Prior RL. The chemistry behind antioxidant capacity assays. J Agric Food Chem 2005;53:1841-56. https://doi.org/10.1021/jf030723c
  27. Xie JT, Shao ZH, Vanden Hoek TL, Chang WT, Li J, Mehendale S, Wang CZ, Hsu CW, Becker LB, Yin JJ, et al. Antioxidant effects of ginsenoside Re in cardiomyocytes. Eur J Pharmacol 2006;532:201-7. https://doi.org/10.1016/j.ejphar.2006.01.001
  28. Naval MV, Gomez-Serranillos MP, Carretero ME, Villar AM. Neuroprotective effect of a ginseng (Panax ginseng) root extract on astrocytes primary culture. J Ethnopharmacol 2007;112:262-70. https://doi.org/10.1016/j.jep.2007.03.010
  29. Kanno S, Ishikawa M, Takayanagi M, Takayanagi Y, Sasaki K. Characterization of hydrogen peroxide-induced apoptosis in mouse primary cultured hepatocytes. Biol Pharm Bull 2000;23:37-42. https://doi.org/10.1248/bpb.23.37
  30. Lu JM, Yao QZ, Chen CY. Ginseng compounds: An update on their molecular mechanisms and medical applications. Curr Vasc Pharmacol 2009;7:293-302. https://doi.org/10.2174/157016109788340767
  31. Ye RD, Han JL, Kong XW, Zhao LZ, Cao R, Rao ZR, Zhao G. Protective effects of ginsenoside Rd on PC12 cells against hydrogen peroxide. Biol Pharm Bull 2008;31:1923-7. https://doi.org/10.1248/bpb.31.1923
  32. He F, Guo R, Wu SL, Sun M, Li M. Protective effects of ginsenoside rb1 on human umbilical vein endothelial cells in vitro. J Cardiovasc Pharmacol 2007;50:314-20. https://doi.org/10.1097/FJC.0b013e3180cab12e
  33. Sohn SH, Kim SK, Kim YO, Kim HD, Shin YS, Yang SO, Kim SY, Lee SW. A comparison of antioxidant activity of Korean white and red ginsengs on $H_2O_2$-induced oxidative stress in HepG2 hepatoma cells. J Ginseng Res 2013;37:442-50. https://doi.org/10.5142/jgr.2013.37.442
  34. Brunelle JK, Chandel NS. Oxygen deprivation induced cell death: An update. Apoptosis 2002;7:475-82. https://doi.org/10.1023/A:1020668923852
  35. Jia L, Zhao YQ, Liang XJ. Current evaluation of the millennium phytomedicineginseng (ii): Collected chemical entities, modern pharmacology, and clinical applications emanated from traditional Chinese medicine. Curr Med Chem 2009;16:2924-42. https://doi.org/10.2174/092986709788803204
  36. Lopez MV, Cuadrado MP, Ruiz-Poveda OM, Del Fresno AM, Accame ME. Neuroprotective effect of individual ginsenosides on astrocytes primary culture. Biochim Biophys Acta 2007;1770:1308-16. https://doi.org/10.1016/j.bbagen.2007.06.008
  37. Shin HS, Yu M, Kim M, Choi HS, Kang DH. Renoprotective effect of red ginseng in gentamicin-induced acute kidney injury. Lab Invest 2014;94:1147-60. https://doi.org/10.1038/labinvest.2014.101
  38. Hwang CR, Lee SH, Jang GY, Hwang IG, Kim HY, Woo KS, Lee J, Jeong HS. Changes in ginsenoside compositions and antioxidant activities of hydroponic-cultured ginseng roots and leaves with heating temperature. J Ginseng Res 2014;38:180-6. https://doi.org/10.1016/j.jgr.2014.02.002
  39. Liu ZQ, Luo XY, Liu GZ, Chen YP, Wang ZC, Sun YX. In vitro study of the relationship between the structure of ginsenoside and its antioxidative or prooxidative activity in free radical induced hemolysis of human erythrocytes. J Agric Food Chem 2003;51:2555-8. https://doi.org/10.1021/jf026228i
  40. Liu ZQ, Luo XY, Liu GZ, Liu QW. Including 4-hydroxyquinoline derivatives into beta-cyclodextrin to form complexes affects their antioxidative effect on freeradical-induced hemolysis of human erythrocytes. QSAR Comb Sci 2003;22:859-64. https://doi.org/10.1002/qsar.200330838
  41. Lee YJ, Kim HY, Kang KS, Lee JG, Yokozawa T, Park JH. The chemical and hydroxyl radical scavenging activity changes of ginsenoside-rb-1 by heat processing. Bioorg Med Chem Lett 2008;18:4515-20. https://doi.org/10.1016/j.bmcl.2008.07.056
  42. Choi WY, Choi WS, Kwon HS, Lee HY. Enhancement of low molecular weight ginsenosides from low-quality ginseng through ultra-high-pressure and fermentation processes. Eur Food Res Technol 2013;237:429-40. https://doi.org/10.1007/s00217-013-2007-9
  43. Kang KS, Yokozawa T, Yamabe N, Kim HY, Park JH. ESR study on the structure and hydroxyl radical-scavenging activity relationships of ginsenosides isolated from Panax ginseng Meyer. Biol Pharm Bull 2007;30:917-21. https://doi.org/10.1248/bpb.30.917
  44. Kim GN, Lee JS, Song JH, Oh CH, Kwon YI, Jang HD. Heat processing decreases amadori products and increases total phenolic content and antioxidant activity of Korean Red Ginseng. J Med Food 2010;13:1478-84. https://doi.org/10.1089/jmf.2010.1076
  45. Wolfe KL, Kang XM, He XJ, Dong M, Zhang QY, Liu RH. Cellular antioxidant activity of common fruits. J Agric Food Chem 2008;56:8418-26. https://doi.org/10.1021/jf801381y
  46. Liao W, Ning Z, Chen L, Wei Q, Yuan E, Yang J, Ren J. Intracellular antioxidant detoxifying effects of diosmetin on 2,2-azobis(2-amidinopropane) dihydrochloride (aaph)-induced oxidative stress through inhibition of reactive oxygen species generation. J Agric Food Chem 2014;62:8648-54. https://doi.org/10.1021/jf502359x
  47. Wen L, Guo X, Liu RH, You L, Abbasi AM, Fu X. Phenolic contents and cellular antioxidant activity of chinese hawthorn “Crataegus pinnatifida”. Food Chem 2015;186:54-62. https://doi.org/10.1016/j.foodchem.2015.03.017

Cited by

  1. Individual and combined antioxidant effects of ginsenoside F2 and cyanidin-3-O-glucoside in human embryonic kidney 293 cells vol.6, pp.84, 2017, https://doi.org/10.1039/c6ra14831j
  2. Individual and combined use of ginsenoside F2 and cyanidin-3-O-glucoside attenuates H2O2-induced apoptosis in HEK-293 cells via the NF-κB pathway vol.7, pp.66, 2017, https://doi.org/10.1039/c7ra04689h
  3. Anti-Wrinkling and Anti-Melanogenic Effect of Pradosia mutisii Methanol Extract vol.20, pp.5, 2017, https://doi.org/10.3390/ijms20051043
  4. Reduced nitrogen supply enhances the cellular antioxidant potential of phenolic extracts through alteration of the phenolic composition in lettuce (Lactuca sativa L.) vol.99, pp.10, 2019, https://doi.org/10.1002/jsfa.9721
  5. Phytochemical profiling and screening of protective effects of Artabotrys odoratissimus on H2O2 induced oxidative stress in HEK-293 cells and erythrocytes vol.167, pp.4, 2020, https://doi.org/10.1080/23818107.2020.1791730
  6. Mitochondrial TXN2 attenuates amyloidogenesis via selective inhibition of BACE1 expression vol.157, pp.4, 2017, https://doi.org/10.1111/jnc.15184