Antioxidant Effects of Gagam-jangwon-hwan(jiajianzhuangyuanwan) on Hydrogen Peroxide-Induced Oxidative Stress in PC 12 Cell Lines

$H_2O_2$로 유도된 산화적 스트레스에 대한 장원환가감방(壯元丸加減方)의 PC 12 cell 에서의 항산화 효과

  • Park, Yong-Hoon (IN AM NeuroScience Research Center, Sanbon Medical Center, Wonkwang University) ;
  • Son, Il-Hong (IN AM NeuroScience Research Center, Sanbon Medical Center, Wonkwang University) ;
  • Lee, Sang-Won (National Institute of Horticultural and Herbal Science) ;
  • Lim, Jung-Hyun (Dept. of neuropsychiatry, college of Oriental Medicine, Wonkwang University) ;
  • Kim, Tae-Heon (Dept. of neuropsychiatry, college of Oriental Medicine, Wonkwang University) ;
  • Lyu, Yeoung-Su (Dept. of neuropsychiatry, college of Oriental Medicine, Wonkwang University) ;
  • Kang, Hyung-Won (Dept. of neuropsychiatry, college of Oriental Medicine, Wonkwang University)
  • 박용훈 (원광대학교 산본병원 인암뇌신경센타) ;
  • 손일홍 (원광대학교 산본병원 인암뇌신경센타) ;
  • 이상원 (국립원예특작과학원) ;
  • 임정현 (원광대학교 한의과대학 한방신경정신과학교실) ;
  • 김태헌 (원광대학교 한의과대학 한방신경정신과학교실) ;
  • 류영수 (원광대학교 한의과대학 한방신경정신과학교실) ;
  • 강형원 (원광대학교 한의과대학 한방신경정신과학교실)
  • Published : 2009.06.30

Abstract

Objectives : Antioxidant effects of Gagam-jangwonhwan(LMK01 and 02) water extract against $H_2O_2$-induced oxidative damage and cell death were investigated in rat pheochromocytoma line PC 12. Methods : The cells were treated with LMK01 and 02 water extract and $H_2O_2$, oxidative damage-inducing materials for 24 h. The cellular viability was assessed by WST-1 assay, oxidative damages of the cells by 8-OHdG quantitation, apoptosis by Hoechst 33342 staining assay and activity of antioxidant enzymes by catalase and glutathione peroxidase assay. Results : 1. LMK01 and LMK02 water extracts improved significantly cell viability in $H_2O_2$-treated groups than $H_2O_2$-alone treated cells 2, LMK02 suppressed significantly oxidative damage in $H_2O_2$-treated groups than $H_2O_2$-alone treated cells but LMK01 didn't. Meanwhile, difference of oxidative damages in conditions treated with LMK01 or LMK02 was not significant, 3. The $H_2O_2$ induced-apoptosis in PC 12 cell lines was inhibited effectively by LMK01 and LMK02, and especially the features of apoptosis were obviously reduced in LMK02-treated cells. 4. LMK01 and LMK02 increased significantly activities of both catalase and glutathione peroxidase than those of $H_2O_2$-alone treated group and moreover, LMK02 showed significantly higher activities than those of LMK01. Conclusions : As shown, LMK01 and LMK02 suppressed $H_2O_2$-induced oxidative damage and cell death in PC 12 cell effectively. And they increased activity of major antioxidant enzymes in PC 12 cell line. Therefore, this study suggests the possibility of clinical usage over oxidative stress-induced neurodegenerative disease such as Alzheimer's disease.

Keywords

References

  1. 김안근, 차은정. 산화적 스트레스상태에서 B16F10 Murine Melanoma 세포의 항산화효소 활성에 대한 흑축 추출물의 효과. 대한약학회지. 2004;48(1):93-8.
  2. Branen, A. S. Toxicology and biochemistry of butylated hydroxyanisole and butylated hydroxytoluene. Journal of the American Oil Chemists' Society. 1975;52(2):59-63. https://doi.org/10.1007/BF02901825
  3. Finkel T, Holbrook NJ. Oxidants, oxidative stress and the biology of ageing. Nature. 2000;408:239-47. https://doi.org/10.1038/35041687
  4. PF Good, P Werner, A Hsu, CW Olanow, DP Perl. Evidence of neuronal oxidative damage in Alzheimer's disease. Am. J. Patho. 1996:149,21-28.
  5. Smith, Mark A. Hirai, Keisuke. Hsiao, Karen. Pappolla, Miguel A. Harris, Peggy L. R. Siedlak, Sandra L. Tabaton, Massimo. Perry, George. Amyloid-[beta] Deposition in Alzheimer Transgenic Mice Is Associated with Oxidative Stress. Journal of Neurochemistry. 1998;70(5):2212-5. https://doi.org/10.1046/j.1471-4159.1998.70052212.x
  6. 윤미영, 김주영, 황지환, 차미란, 이미라, 조경진, 박해룡. 석곡 MeOH 추출물이 H2O2에 의한 신경세포 보호효과에 미치는 영향. 한국응용생명화학회. 2007;50(1):63-7.
  7. 성금수, 전철, 권용훈, 김경현, 장재철. 홍삼추출물 투여가 생쥐간에서 항산화 효소 활성과 지질과산화에 미치는 효과. 고려인삼학회지. 2000;24(1):29-34.
  8. 강우석, 김정한, 박은주, 윤광로. 울금 에탄올 추출물의 항산화 활성 비교. 한국식품과학회지. 1998;30(2):266-71.
  9. 강형원, 박진성, 유영수. 釣鉤藤이 산소자유기에 의하여 손상된 培養脊髓感覺神經節細胞에 미치는 영향에 관한 연구. 동의신경정신과학회지. 2000;11(1):1-18.
  10. 박종운, 한상혁, 김도환, 문병순. H2O2에 의한 배양심근세포고사에 미치는 四物湯의 방어효과. 대한한의학회지. 2001;22(4):58-68.
  11. 임준모. 이민구, 윤종민, 이인, 문병순. 산화적 손상에 의한 Neuro 2A 치매모델에서 石 菖蒲遠志散의 방어효과. 대한한의학회지. 2005;26(1):161-73.
  12. 許俊. 東醫寶鑑. 서울:法仁文化社. 1999:189-91.
  13. 강형원, 김상태, 손형진, 한평림, 조형권, 이영재, 류영수. LMK02의 품질규격화와 $A\beta$ 올리고머에 의해 유도된 흰쥐해마 H19-7세 포주에 미치는 항치매효과. 동의생리병리학회지. 2009;23(2):397-404.
  14. 김상태, 강형원, 한평림, 조형권, 김태헌, 류영수, 손형진. 壯元丸加減方인 LMK02가 아밀로이드 前驅蛋白質으로 形質轉換된 초파리에 미치는 효과. 동의신경정신과학회지. 2008;19(2):151-63.
  15. Ames BN. Shigenaga MK, Hagen TM. Oxidants, antioxidants, and the degenerative diseases of aging. Proc Natl Acad Sci U S A. 1993;90(17):7915-22. https://doi.org/10.1073/pnas.90.17.7915
  16. Ames BN. Dietary carcinogens and anticarcinogens. Oxygen radicals and degenerative diseases. Science. 1983;221:1256-64. https://doi.org/10.1126/science.6351251
  17. MF Beal. Mechanisms of excitotoxicity in neurologic diseases. The FASEB J. 1992;6:3338-44.
  18. Choi DW. Excitotoxic cell death. J. Neurobiol. 1992;23:1261-76 . https://doi.org/10.1002/neu.480230915
  19. Coyle JT, Puttfarcken P. Oxidative stress. glutamete. and neurodegenerative disoders. Science. 1993;262:689-95. https://doi.org/10.1126/science.7901908
  20. Markesbery WR. Oxidative stress hypothesis in Alzheimer's disease. Free Radic Biol Med. 1997;23:134-47. https://doi.org/10.1016/S0891-5849(96)00629-6
  21. Schippling S, Kontush A, Arlt S, Buhmann C, Sturenbug HJ, Mann U, Muller-Thomsen T, Beisiegel U. Increased lipoprotein oxidation in Alzheimer's disease. Free Radic Biol Med. 2000;28:351-60. https://doi.org/10.1016/S0891-5849(99)00247-6
  22. 김건진, 정대규. 장원단이 CT105와 $A\beta$로 유발된 치매병태모델에 미치는 영향. 동의신경정신과학회지. 2006;17(2):91-122.
  23. 강형원, 김상태, 이종화, 김태헌, 손형진, 한평림, 류영수. 壯元丸加減方 水抽出物이 아밀로이드 전구단백질으로 유도된 생쥐의 신경아세포주에서의 항치매 효과. 동의신경정신과학회지. 2007;18(2):13-24.
  24. 龔廷賢. 增補萬病回春. 서울:一中社. 1994:229-30.