PC12 Cell Protective Effects of Broccoli (Brassica oleracea var. italica) Leaf Fraction against H2O2-induced Oxidative Stress

H2O2로 유발된 산화적 스트레스에 대한 브로콜리(Brassica oleracea var. italica) 잎 분획물의 PC12 cell 보호 효과

  • Park, Seon Kyeong (Division of Applied Life Science, Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Jin, Dong Eun (Division of Applied Life Science, Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Park, Chang Hyeon (Division of Applied Life Science, Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Seung, Tae Wan (Division of Applied Life Science, Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Choi, Sung-Gil (Division of Applied Life Science, Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Heo, Ho Jin (Division of Applied Life Science, Institute of Agriculture and Life Science, Gyeongsang National University)
  • 박선경 (경상대학교 응용생명과학부, 농업생명과학연구원) ;
  • 진동은 (경상대학교 응용생명과학부, 농업생명과학연구원) ;
  • 박창현 (경상대학교 응용생명과학부, 농업생명과학연구원) ;
  • 승태완 (경상대학교 응용생명과학부, 농업생명과학연구원) ;
  • 최성길 (경상대학교 응용생명과학부, 농업생명과학연구원) ;
  • 허호진 (경상대학교 응용생명과학부, 농업생명과학연구원)
  • Received : 2014.04.03
  • Accepted : 2013.05.26
  • Published : 2014.08.31


To examine the physiological effects of broccoli (Brassica oleracea var. italica) leaf, the bioavailable compounds in broccoli leaf extract, and its in vitro neuroprotective effects against $H_2O_2$-induced oxidative stress were examined in this study. The chloroform fraction of broccoli leaf extract had the highest total phenolic content of all the fraction than others, and the highest 2,2"-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical-scavenging activity and malondialdehyde (MDA) inhibitory effect. Intracellular reactive oxygen species (ROS) accumulation resulting in $H_2O_2$-treated in PC12 cells was significantly lower when the chloroform fraction was present in the medium compared to that in PC12 cells treated with $H_2O_2$ alone. In a cell viability assay performed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), the chloroform fraction showed protective effects against $H_2O_2$-induced neurotoxicity and inhibited lactate dehydrogenase (LDH) release into the medium. High-performance liquid chromatography (HPLC) analysis showed that ferulic acid was the predominant phenolic compound in chloroform fraction of broccoli leaf.


broccoli;Brassica oleracea var. italica;PC12 cell;ferulic acid


Supported by : 한국연구재단


  1. Park SH, Hwang HS, Han JH. Development of drink from composition with medicinal plants and evaluation of its physiological function. Korean J. Nutr. 37: 364-372 (2004)
  2. Jeong EJ, Kim JW, Kim YC. Rhus verniciflua stokes attenuates glutamate-induced neurotoxicity in primary cultures of rat cortical cells. Nat. Prod. Sci. 14: 156-160 (2008)
  3. Fridovich, I. Biological effects of the superoxide radical. Arch. Biochem. Biophys. 247: 1-11 (1986)
  4. Heo HJ, Kim DO, Choi SJ, Shin DH and Lee CY. Potent inhibitory effect of flavonoids in Scutellaria baicalensis on amyloid a protein-induced neurotoxicity. J. Agr. Food Chem. 52: 4128-4132 (2004)
  5. Bidchol AM, Wilfred A, Abhijna P and Harish R. Free radical scavenging activity of aqueous and ethanolic extract of Brassica oleracea L. var. italic. Food Bioprocess Technol. 4: 1137-1143 (2011)
  6. Nohl H and Jordan, W. The mitochondrial site of superoxide formation. Biochem. Biophy. Res. Commun. 138: 533-539 (1986)
  7. Yoon MY, Lee BB, Kim JY, Kim YS, Park EJ, Lee SC, Park HR. Antioxidant activity and neuroprotective effect of psoralea corylifolia linne extracts. Korean J. Pharmacol. 38: 84-89 (2007)
  8. Matucsheski NV, Juvik JA, Jeffery EH. Heating decreases epithiospecifier protein activity and increases sulforaphane formation in broccoli. Phytochemistry 65: 1273-1281 (2004)
  9. Stoewsand G.S. Bioactive organosulfur phytochemicals in Brassica oleracea vegetables-a review, Food Chem. Toxicol. 33: 537 (1995)
  10. Caudhary A, Sharma U, Vig AP, Singh B, Arora S. Free radical scavenging, antiproliferative activities and profiling of variations in the level of phytochemicals in different parts of broccoli (Brassica oleracea italica). Food Chem. 148: 373-380 (2014)
  11. Lee HS, Park YW. Antioxidant activity and antibacterial activities from different parts of broccoli extracts under high temperature. J. Korean Soc. Food Sci. Nutr. 34: 759-764 (2005)
  12. Kim CH, Cho KR. Quality characteristics of sponge cakes made with different quantities of broccoli powder. Korean J. Food Sci. Technol. 42: 459-467 (2010)
  13. Arnaiz E, Bernal J, Martin MT, Nozal MJ, Bernal JL, Toribio L. Supercritical fluid extraction of free amino acids from broccoli leaves. J. Chromatogr. A 1250: 49-53 (2012)
  14. Kim, DO, Jeong SW, Lee CY. Antioxidant capacity of phenolic phytochemical from various cultivars of plums. Food Chem. 81: 321-326 (2003)
  15. Kwak JH, Choi GN, Pack JH, Kim JH, Jeong HR, Jeong CH, Heo HJ. Antioxidant and neuronal cell protective effect of purple sweet potato extract. J. Agric. Life Sci. 44: 57-66 (2010)
  16. Chang ST, Wu JH, Wang SY, Kang PL,Yang NS, Shyur, LF. Antioxidant Activity of Extracts from Acacia confusa Bark and Heartwood. J. Agr. Food Chem. 49: 3420-3424 (2001)
  17. Kim JK, Choi SJ, Cho HY, Hwang HJ, Kim YJ, Lim T, Kim CJ, Kim HK, Peterson S, Shin DH. Protective effects of kamepferol(3,4',5,7-tetrahydroxyflavone) against amyloid beta peptide($A{\beta}$)-induced neurotoxicity in ICR mice. Biosci. Biotechnol. Biochem. 74: 397-401 (2010)
  18. Yoo KM, Kim DO, Lee CY. Evaluation of different methods of antioxidant measurement. Food Sci. Biotechnol. 16: 177-182 (2007)
  19. Kim SS, Park RY, Jeon HJ, Kwon YS, Chun W. Neuroprotective effects of 3,5-dicaffeoylquinic acid on hydrogen peroxide-induced cell death in SH-SY5Y cells. Phytother. Res. 19: 243-245 (2005)
  20. Heo HJ, Cho HY, Hong BS, Kim HK, Kim EK, Kim BK, Shin DH. Protective effect of 4',5-dihydroxy-3',6,7-trimethoxyflavone from Artemisia asiatica against $A{\beta}$-induced oxidative stress on PC12 cells. Amyloid 8: 194-201 (2001)
  21. Kim KH, Kim HJ, Byun MW, Yook HS. Antioxidnat and antimicrobial activities of ethanol extract from six vegetables containing different sulfur compounds. J. Korean Soc. Food Sci. Nutr. 41: 577-583 (2012)
  22. Re R, Plellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Bio. Med. 26: 1231-1237 (1999)
  23. Uchida K, Stadtman ER. Covalent attachment of 4-hydroxynonenal to glyceraldehyde-3-phosphate dehydrogenase A possible involvement of intra and intermolecular cross-linking reaction. J. Biol. Chem. 268: 6388-6393 (1993)
  24. Zhao B. Natural antioxidants protect neurons in Alzheimer's disease and Parkinson's disease. Neurochem. Res. 34: 630-638 (2009)
  25. Hurst RD, Wells RW, Hurst SM, McGhie TK, Cooney JM, Jensen DJ. Blueberry fruit polyphenolics suppress oxidative stress-induced skeletal muscle cell damage in vitro. Mol. Nutr. Food Res. 54: 353-363 (2010)
  26. Kim HY, Lee YA, Cho EJ. Free radical scavenging effect and protective activity from oxidative stress of broccoli flowers and sprouts. CNU J. Agric. Sci. 39: 81-86 (2012)
  27. Heo HJ, Choi SJ, Choi SG, Shin DH, Lee JM, Lee CY. Effect of banana, orange, and apple on oxidative stress-induced neurotoxicity in PC12cells. J. Food Sci. 73: H28-32 (2008)
  28. Zhao Z, Moghadasian MH. Chemistry, natural sources, dietary intake and pharmacokinetic properties of ferulic acid: A review. Food Chem. 109: 691-702 (2008)
  29. Kanski J, Aksenova M, Stoyanova A, Butterfield A. Ferulic acid antioxidant protection against hydroxyl and peroxyl radical oxidation in snaptosomal and neuroal cell culture systems in vitro: structure-activity studies. J. Nutr. Biochem. 13: 273-281 (2002)

Cited by

  1. Antiamnesic Effect of Broccoli (Brassica oleraceavar.italica) Leaves on Amyloid Beta (Aβ)1–42-Induced Learning and Memory Impairment vol.64, pp.17, 2016,