DOI QR코드

DOI QR Code

Cirsium japonicum Extracts Show Antioxidant Activity and PC12 Cell Protection against Oxidative Stress

좁은잎 엉겅퀴 추출물의 산화방지 활성 및 산화적 스트레스에 대한 PC12 세포 보호효과

  • Jang, Miran (Plant Resources Research Institute, Duksung Women's University) ;
  • Kim, Gun-Hee (Plant Resources Research Institute, Duksung Women's University)
  • 장미란 (덕성여자대학교 식물자원연구소) ;
  • 김건희 (덕성여자대학교 식물자원연구소)
  • Received : 2015.12.08
  • Accepted : 2016.03.23
  • Published : 2016.04.30

Abstract

The phenolic compounds, antioxidant activity and neuronal cell protective effect of Cirsium japonicum extract were evaluated in this study. High performance liquid chromatography mass analysis showed that C. japonicum was composed of chlorogenic acid, linarin, and pectolinarin. C. japonicum extract showed its antioxidant activity with half-maximal inhibitory concentrations of 567 and $130{\mu}g/mL$ by DPPH and ABTS radical scavenging activity, respectively. The total antioxidant capacities of C. japonicum via DPPH, ABTS, and FRAP assays were 11.32, 100.15, and $12.76{\mu}g/mL$ trolox equivalents, respectively. In addition, the neuroprotective effect of C. japonicum extract was investigated by measuring cell viability via MTT, LDH and DCF-DA assay using $H_2O_2-damaged$ PC12 cells. C. japonicum extract showed neuronal cell protective effects in a dose-dependent manner. These results indicated that C. japonicum extract has potent antioxidant and neuronal protective effects. Therefore, C. japonicum can be regarded as an effective and safe functional food resource as natural antioxidants, and may decrease the risk of neurodegenerative disorders.

HPLC/MS를 이용하여 국내 자생 엉겅퀴(Cirsium japonicum)의 페놀성 화합물을 분석한 결과 제주지역 엉겅퀴가 가장 많은 페놀성 화합물을 함유하고 있었으며, chlorogenic acid가 73.15 mg/g dry weight, linarin이 76.67 mg/g dry weight 그리고 pectolinarin이 12.98 mg/g dry weight으로 확인되었다. 엉겅퀴의 기능성 식품으로서의 가치를 확인하기 위하여 산화방지 활성 및 신경세포 보호효과를 평가한 결과 DPPH, ABTS 및 FRAP assay에서 엉겅퀴의 강력한 산화방지 활성이 나타났다. 엉겅퀴 추출물이 DPPH 및 ABTS 라디칼의 50%를 저해하는 농도는 각각 $567{\mu}g/mL$$130{\mu}g/mL$으로 나타났다. DPPH, ABTS, FRAP법을 통한 총산화방지능은 각각 11.32, 100.15, $12.76{\mu}g/mL$ trolox equivalents 나타났다. 과산화수소로 산화적 손상을 유도한 PC12 세포에 대하여 MTT와 LDH assay를 이용하여 세포생존률을 측정한 결과 농도 의존적으로 세포 보호 활성이 나타났으며, 마찬가지로 활성산소종 생성률을 측정한 결과 농도 의존적으로 활성산소종 생성이 감소되어 세포 보호활성이 확인되었다. 본 연구를 통하여 엉겅퀴의 우수한 산화방지 활성 및 신경세포 보호효과가 검증되었다. 따라서 엉겅퀴는 안전한 식품 재료로서 꾸준히 섭취하였을 때 천연 산화방지제로 작용하여 신경퇴행을 예방함으로써 알츠하이머병, 파킨슨병 및 헌팅턴병 등의 질병 위험을 줄일 수 있을 것으로 판단된다.

Keywords

References

  1. Lu YH, Su MY, Huang HY, Li L, Yuan CG. Protective effects of the citrus flavanones to PC12 cells against cytotoxicity induced by hydrogen peroxide. Neucosci. Lett. 484: 6-11 (2010) https://doi.org/10.1016/j.neulet.2010.07.078
  2. Esposito E, Rotilio D, Matteo VD, Giulio CD, Cacchio M, Algeri S. A review of specific dietary antioxidants and the effects on biochemical mechanisms related to neurodegenerative processes. Neurobiol. Aging 23: 719-735 (2002) https://doi.org/10.1016/S0197-4580(02)00078-7
  3. Jeong CH, Kwak JH, Kim JH, Choi GN, Kim DO, Heo HJ. Neuronal cell protective and antioxidant effects of phenolics obtained from Zanthoxylum piperitum leaf using in vitro model system. Food Chem. 125: 417-422 (2011) https://doi.org/10.1016/j.foodchem.2010.09.022
  4. Jang MR, Park HJ, Hong EY, Kim GH. Comparison of the antibacterial activity of domestic Cirsium japonicum collected from different regions. Korean J. Food Cook. Sci. 30: 278-283 (2014) https://doi.org/10.9724/kfcs.2014.30.3.278
  5. Chon SU, Kim YM, Kim DK, He BG, Cho JY. Phytotoxic effect, DPPH radical scavenging activity and chlorogenic acid level of methanol extracts from aerial parts of several Korean salad plants. Korean J. Plant Resources 19: 405-410 (2006)
  6. Ganzera M, Pocher P, Stuppner H. Differentiation of Cirsium japonicum and C. setosum by TLC and HPLC-MS. Phytochem. Anal. 16: 205-209 (2005) https://doi.org/10.1002/pca.846
  7. Ge H, Turhong M, Abudkrem M, Tang Y. Fingerprint analysis of Cirsium japonicum DC. using high performance liquid chromatography. J. Pharm. Anal. 3: 278-284 (2013) https://doi.org/10.1016/j.jpha.2012.12.004
  8. Jeong DM, Jung HA, Choi JS. Comparative antioxidant activity and HPLC profiles of some selected Korean thistles. Arch. Pharm. Res. 31: 28-33 (2008) https://doi.org/10.1007/s12272-008-1116-7
  9. Kim SJ, Kim GH. Identification for flavones in different parts of Cirsium japonicum. J. Food. Sci. Nutr. 8: 330-335 (2003) https://doi.org/10.3746/jfn.2003.8.4.330
  10. Greenlee H, Abascal K, Yarnell E, Ladas E. Clinical applications of Silybum marianum in oncology. Integr. Cancer. Ther. 6: 158-165 (2007) https://doi.org/10.1177/1534735407301727
  11. Jang MR, Hong EY, Cheong JH, Kim GH. Antioxidative components and activity of domestic Cirsium japonicum extract. J. Korean Soc. Food Sci. Nutr. 41: 739-744 (2012) https://doi.org/10.3746/jkfn.2012.41.6.739
  12. Lee JH, Choi SI, Lee YS, Kim GH. Antioxidant and anti-inflammatory activities of ethanol extract from leaves of Cirsium japonicam. Food Sci. Biotech. 17: 38-45 (2008)
  13. Yin Y, Heo SI, Wang MH. Antioxidant and anticancer activities of methanol and water extracts from leaves of Cirsium japonicam. J. Appl. Biol. Chem. 51: 160-164 (2008) https://doi.org/10.3839/jabc.2008.030
  14. Chon SU, Kim TS, Shin JS, Boo HO. Antioxidant activities of methanol extracts from root parts of Korean salad plants. Korean J. Plant Res. 21: 413-419 (2008)
  15. Yin J, Heo SI, Wang MH. Antioxidant and antidiabetic activities of extracts from Cirsium japonicam roots. Nutr. Res. Pract. 2: 247-251 (2008) https://doi.org/10.4162/nrp.2008.2.4.247
  16. Kim EM, Won SI. Functional Composition and antioxidative activity from different organs of native Cirsium and Carduus genera. Korean J. Food Cook. Sci. 25: 406-414 (2009)
  17. Mok JY, Kang HJ, Cho JK, Jeon IH, Kim HS, Park JM, Jeong SI, Shim JS, Jang SI. Antioxidative and anti-inflammatory effects of extracts from different organs of Cirsium japonicum var. ussuriense. Kor. J. Herbol. 26: 39-47 (2011)
  18. Lee HK, Kim JS, Kim NY, Kim MJ, Park SU, Yu CY. Antioxidant, antimutagenicity and anticancer activities of extracts from Cirsium japonicum var. ussuriense KITAMURA. J. Med. Crop Sci. 11: 53-61 (2003)
  19. Liu Y, Tia X, Gou L, Sun L, Ling X, Yin X. Luteolin attenuates diabetes-associated cognitive decline in rats. Brain Res. Bull. 94: 23-29 (2013) https://doi.org/10.1016/j.brainresbull.2013.02.001
  20. Han JY, Ahn SY, Kim CS, Yoo SK, Kim SK, Kim HC, Hong JT, Oh KW. Protection of apigenin against kainate-induced excitotoxicity by anti-oxidative effects. Biol. Pharm. Bull. 35: 1440-1446 (2012) https://doi.org/10.1248/bpb.b110686
  21. Li G, Min BS, Zheng C, Lee J, OW SR, Ahn KS, Lee BK. Neuroprotective and free radical scavenging activities of phenolic compounds from Hovenia dulcis. Arch. Pharm. Res. 28: 804-809 (2005) https://doi.org/10.1007/BF02977346
  22. Braca A, Politi M, Sanogo R, Sanou H, Morelli I, Pizza C, Tommasi ND. Chemical composition and antioxidant activity of phenolic compounds from wild and cultivated Sclerocarya birrea (Anacardiaceae) leaves. J. Agr. Food Chem. 51: 6689-6695 (2003) https://doi.org/10.1021/jf030374m
  23. Wojdylo A, Oszmia ski J, Czemerys R. Antioxidant activity and phenolic compounds in 32 selected herbs. Food Chem. 105: 940- 949 (2007) https://doi.org/10.1016/j.foodchem.2007.04.038
  24. Heo HJ, Cho HY, Hong B, Kim HK, Kim EK, Kim BG, Shin DH. Protective effect of 4',5-dihydroxy-3',6,7-trimethoxyflavone from Artemisia asiatica against A${\beta}$-induced oxidative stress in PC12 cells. Amyloid 8: 194-201 (2001) https://doi.org/10.3109/13506120109007362
  25. Prior RL, Xu X, Schaich K. Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements. J. Agr. Food Chem. 53: 4290-4302 (2005) https://doi.org/10.1021/jf0502698
  26. Kwak JH, Choi GN, Park JH, Kim JH, Jeong HR, Jeong CH, Heo HJ. Antioxidant and neuronal cell protective effect of purple sweet potato extract. J. Agr. Life Sci. 44: 57-66 (2010)
  27. Rice-Evans CA, Miller NJ, Paganga G. Paganga Structure-antioxidant activity relationships of flavonoids and phenolic acids. Free Radical Bio. Med. 20: 933-956 (1996) https://doi.org/10.1016/0891-5849(95)02227-9
  28. Choi DY, Lee YJ, Hong JT, Lee HJ. Antioxidant properties of natural polyphenols and their therapeutic potentials for Alzheimer's disease. Brain Res. Bull. 10: 144-153 (2011)
  29. Heo HJ, Choi SJ, Choi SG, Shin DH, Lee JM Lee CY. Effect of banana, orange, and apple on oxidative stress-induced neurotoxicity in PC12 cells. J. Food Sci. 73: H28-32 (2008) https://doi.org/10.1111/j.1750-3841.2007.00632.x
  30. Lobner D. Comparison of the LDH and MTT assays for quantifying cell death: validity for neuronal apoptosis? J. Neurosci. Meth. 96: 147-152 (2000) https://doi.org/10.1016/S0165-0270(99)00193-4
  31. Zhao B. Natural antioxidants protect neurons in Alzheimer's disease and Parkinson's disease. Neurochem. Res. 34: 630-638 (2009) https://doi.org/10.1007/s11064-008-9900-9
  32. Li Y, Shi W, Li Y, Zhou Y, Hua X, Song C, Ma H, Wang C, Li Y. Neuroprotective effects of chlorogenic acid against apoptosis of PC12 cells induced by methylmercury. Environ. Toxicol. Phar. 26: 13-21 (2008) https://doi.org/10.1016/j.etap.2007.12.008
  33. Mikami Y, Yamazawa M. Chlorogenic acid, a polyphenol in coffee, protects neurons against glutamate neurotoxicity. Life Sci. 139: 69-74 (2015) https://doi.org/10.1016/j.lfs.2015.08.005
  34. Lou H, Fan P, Perez RG, Lou H. Neuroprotective effects of linarin through activation of the PI3K/Akt pathway in amyloid-${\beta}$- induced neuronal cell death. Bioorgan. Med. Chem. 19: 4021-4027 (2011) https://doi.org/10.1016/j.bmc.2011.05.021