Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Cytoprotective Effects and Gene Expression Patterns Observed Based on the Antioxidant Activity of Lonicera japonica Extract

금은화 추출물의 항산화 효과를 통한 세포 보호효과 및 유전자 발현 양상

  • Cho, Won June (Department of Biological Science and the Research Institute for Basic Sciences, Hoseo University) ;
  • Yoon, Hee Seung (Department of Biological Science and the Research Institute for Basic Sciences, Hoseo University) ;
  • Kim, Yong Hyun (Department of Biology, Soonchunhyang University) ;
  • Kim, Jung Min (NAR Center, Inc., Daejeon Oriental Hospital of Daejeon University) ;
  • Yoo, Il Jae (Institute of Nanoproduct Safety Research, Hoseo University) ;
  • Han, Man-Deuk (Department of Biology, Soonchunhyang University) ;
  • Bang, In Seok (Department of Biological Science and the Research Institute for Basic Sciences, Hoseo University)
  • 조원준 (호서대학교 생명과학과) ;
  • 윤희승 (호서대학교 생명과학과) ;
  • 김용현 (순천향대학교 생명과학과) ;
  • 김정민 ((주)엔에이알센터 DNA칩 분석팀) ;
  • 유일재 (호서대학교 나노제품안전성연구센타) ;
  • 한만덕 (순천향대학교 생명과학과) ;
  • 방인석 (호서대학교 생명과학과)
  • Received : 2013.07.01
  • Accepted : 2013.08.20
  • Published : 2013.08.30
Download PDF
⟨ Previous Next ⟩

Abstract

In this study, based on the antioxidative effects in organic solvent fractions obtained from the main methanolic extract of L. japonica, the protective cellular effects and gene expression patterns of ethyl acetate fractions on $H_2O_2$-induced Raw 264.7 cell death ($IC_{50}$) were analyzed. The antioxidant activity of the fractions measured using DPPH free radical scavenging activity increased in a dose-dependent manner, and the $ED_{50}$ exhibited the highest $39.56{\mu}g/ml$ in the ethyl acetate fraction. In addition, the ethyl acetate fractions' cell viability on $H_2O_2$-induced Raw 264.7 cell damage increased in a concentration-dependent manner, showed a visible cell survival rate of 82.49% at a concentration of $100{\mu}g/ml$. The gene expression patterns related to the ethyl acetate fractions' cytoprotective effect in $H_2O_2$-induced Raw 264.7 cell damage presented similar patterns to those of BHA. In comparative analysis for antioxidant activity-related genes affected by ethyl acetate fractions and BHA in $H_2O_2$-induced Raw 264.7 cells, both ethyl acetate fractions and BHA showed very similar gene expression patterns, but the gene expression level of the heme oxygenase 1 (Hmox1) gene making antioxidant enzymes in cells was four times higher in ethyl acetate fractions than BHA. In inflammation-related genes in $H_2O_2$induced Raw 264.7 cells, the T-box transcription factor (Tbx21) gene was expressed about two times more frequently in the ethyl acetate fraction treatment group, while it was expressed half as frequently in the BHA treatment group.

금은화의 메탄올 추출에 의한 유기 용매별 분획에서 항산화 효과를 근거로 에틸 아세테이트 분획에서 $H_2O_2$로 유도된 RAW 264.7 세포독성에 대한 세포보호 효과와 유전자 발현 양상을 분석하였다. 용매별 분획의 항산화 활성은 시료의 농도가 증가할수록 DPPH에 대한 전자공여능도 증가하였으며, $ED_{50}$은 에틸 아세테이트 분획에서 $39.56{\mu}g/ml$로 가장 높게 나타났다. 또한 $H_2O_2$에 의해 유도된 Raw 264.7 세포사멸(($IC_{50}$)에 대하여 에틸 아세테이트 분획은 농도 의존적으로 유의적인 세포 생존율과, $100{\mu}g/ml$의 농도에서 82.49%의 세포보호 효과를 나타내었다. 한편 $IC_{50}$의 $H_2O_2$ 독성에 대한 에틸 아세테이트 분획의 Raw 264.7 세포보호 효과에 관련된 유전자 발현의 패턴은 BHA와 유사한 것으로 나타났다. 항산화 관련 유전자군의 비교 분석에서는 에틸 아세테이트 분획과 BHA는 매우 유사한 유전자 발현 양상을 보였으나, 세포 내 주요 항산화 효소를 만드는 유전자 중 하나인 heme oxygenase 1 (Hmox1) 유전자의 발현 정도가 에틸 아세테이트 분획에서 4 배 이상 증가하였다. 항염증 관련 유전자군의 비교에서는 T-box transcription factor (Tbx21)가 BHA 처리군에서는 2 배 감소하였으나 에틸 아세테이트 분획의 처리군에서는 오히려 약 2 배 증가한 것으로 나타났다.

Keywords

References

  1. Blois, M. S. 1958. Antioxidant determination by the use of a stable free radical. Nature 26, 1199-1200.
  2. Borenfreund, E. and Puerner, J. A. 1984. A simple quantitative procedure using monolayer culture for cytotoxicity assay (HTD/NR-90). J Tissue Cult Methods 9, 7-9.
  3. Chun, H. J., Hwang, S. G., Lee, S. J., Baek, S. H., Jeon, B. H. and Woo, W. H. 2002. Inhibitory effects of butyl alcohol extract from Caesalpina sappan L. on melanogenesis in melan- a cells. Korean J Pharmacogn 33, 130-136.
  4. Chung, K. J., Jung, H. J., Jung, S. K. and Rhee, H. K. 2005. Lonicerae Flos contributes to the chemotaxis of eosinophils and secretion of cytokines in A549 human epithelial cells. Korean J Orient Int Med 26, 129-142.
  5. Davies, K. F. and Goldberg, A. L. 1987. Proteins damaged by oxygen radicals are rapidly degraded in extracts of red blood cells. J Biol Chem 262, 8227-8261.
  6. Fubini, B. 1998. Surface chemistry and quartz hazard. Ann Occup Hyg 42, 521-530.
  7. Gordon, M. H. 1996. Dietary antioxidants in disease prevention. Nat Prod Rep 13, 265-273. https://doi.org/10.1039/np9961300265
  8. Kim, I. L., Kim, H. C., Kook, Y. B., Park, S. J., Park, Y. G., Park, G. H., Seo, B. I., Seo, Y. B., Song, H. J., Shin, M. K., Lee, Y. J., Lee, Y. C., Lee J. H., Lim, G. H., Cho, S. I., Jeong, J. K., Zoo, Y. S. and Choi, H. Y. 2007. Herbal medicine. pp. 240-242, Young lim publishing Co., Seoul, Korea.
  9. Kim, Y., Yang, S. Y., Oh, Y. S., Lee, J. W., Lee, Y. K. and Park, Y. C. 2010. Research trends of Lonicera japonica over the last 10 year. J Daejeon Orient Med 19, 17-23.
  10. Kumaran, A. and Karunakaran, R. J. 2006. Antioxidant and free radical scavenging activity of an aqueous extract of Coleus aromaticus. Food Chem 97, 109-114. https://doi.org/10.1016/j.foodchem.2005.03.032
  11. Lee, Y. S. and Jang, S. I. 2010. Study on the anti-inflammatory effects of remedy prescripted with Lonicerae Flos and scutellariae radix in U937 cells. Korean J Orient Med Prescrip 18, 121-132.
  12. Looi, M. L., Mohd Dali, A. Z., Md Ali, S. A., Wan Ngah, W. Z. and Mohd Yusof, Y. A. 2008. Oxidative damage and antioxidant status in patients with cervical intraepithelial neoplasia and carcinoma of the cervix. Eur J Cancer Prev 17, 555-560. https://doi.org/10.1097/CEJ.0b013e328305a10b
  13. Lrson, R. A. 1988. The antioxidents of higher plants. Phytochemistry 27, 969-978. https://doi.org/10.1016/0031-9422(88)80254-1
  14. Maeura, Y., Weisburger, J. H. and Williams, G. 1984. Dosedependent reduction of N-2-fluorenylacetamide-induced liver cancer and enhancement of bladder cancer in rats by butylated hydroxytoluene. Cancer Res 44, 1604-1610.
  15. Martinez, G. R., Loureiro, A. P. M., Marques, S. A., Miyamoto, S., Yamaguchi, L. F., Onuki, J., Almeida, E. A., Garcia, C. C. M., Barbosa, L. F., Medeiros, M. H. G. and Di Mascio, P. 2003. Oxidative and alkylating damage in DNA. Mutat Res 544, 115-127. https://doi.org/10.1016/j.mrrev.2003.05.005
  16. Piantadosi, C. A., Withers, C. M., Bartz, R. R., MacGarvey, N. C., Fu, P., Sweeney, T. E., Welty-Wolf, K. E. and Suliman, H. B. 2011. Heme oxygenase-1 couples activation of mitochondrial biogenesis to anti-inflammatory cytokine expression. J Biol Chem 286, 16374-16385. https://doi.org/10.1074/jbc.M110.207738
  17. Pitchumoni, S. S. and Doraiswamy, P. M. 1998. Current status of antioxidant therapy for alzheimers disease. J Am Geriatr Soc 46, 1566-1572.
  18. Rehman, Z., Habbib, F. and Shah, W. H. 2004. Utilization of potatopeels extract as a natural anti-oxidant in bean oil. Food Chem 85, 215-220. https://doi.org/10.1016/j.foodchem.2003.06.015
  19. Shin, J. H. and Yoo, S. K. 2012. Antioxidant properties in microbial fermentation products of Lonicera japonica Thunb. Extract J East Asian Soc Dietary Life 22, 95-102.
  20. Szabo, S. J., Kim, S. T., Costa, G. L., Zhang, X., Fathman, C. G. and Glimcher, L. H. 2000. A novel transcription factor, T-bet, direct Th1 lineage commitment. Cell 100, 655-669. https://doi.org/10.1016/S0092-8674(00)80702-3
  21. Valko, M., Leibfritz, D., Moncol, J., Cronin, M. T., Mazur, M. and Telser, J. 2007. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 39, 44-84. https://doi.org/10.1016/j.biocel.2006.07.001
  22. Valko, M., Rhodes, C. J., Moncol, J., Izakovic, M. and Mazur, M. 2006. Free radicals, metals and antioxidants in oxidative stress induced cancer. Chem Biol Interact 160, 1-40. https://doi.org/10.1016/j.cbi.2005.12.009
  23. Yun, K. J. and Lee, E. Y. 2012. Effects of hot aqueous and ethanol extract from Lonicera japonica Flos on NO and PGE2 in macrophage. J Korean Acupunct Moxa Soc 29, 67-74.

Cited by

  1. Antioxidant and Protective Effects of Leonurus sibiricus L. Extract on Ultraviolet B (UVB)-induced Damage in Human Keratinocytes vol.29, pp.1, 2016, https://doi.org/10.7732/kjpr.2016.29.1.011
  2. Analgesic Effects of ChondroT in Collagenase-induced Osteoarthritis Rat Model vol.26, pp.3, 2016, https://doi.org/10.18325/jkmr.2016.26.3.17
  3. The antioxidative and cytoprotective effect of Lonicerae japonicae Flos water extracts on the ultraviolet(UV)B-induced human HaCaT keratinocytes vol.29, pp.6, 2014, https://doi.org/10.6116/kjh.2014.29.6.63.