Journal of Sasang Constitutional Medicine (사상체질의학회지)

The Society Of Sasang Constitutional Medicine (사상체질의학회)
권호 표지

Protection effect of New-Yeolda-Hanso tang against $\beta$-Amyloid Induced Cytotoxicity in NGF-differentiated PC12 Cells

$\beta$-Amyloid로 유도된 신경독성에 대한 열다한소탕(熱多寒少湯) 가감방(加感方)의 항(抗)치매효과

  • Bae, Na-Young (Dept. of Sasang Constitution Medicine, College of Oriental Medicine, Daejeon Univ.) ;
  • Yang, Hyun-Ok (Natural Product Research Center, Korea Institute of Science and Technology) ;
  • Ahn, Taek-Won (Dept. of Sasang Constitution Medicine, College of Oriental Medicine, Daejeon Univ.)
  • 배나영 (대전대학교 한의과대학 사상체질과) ;
  • 양현옥 (한국과학기술 연구원 강릉분원 천연물 소재 연구센터) ;
  • 안택원 (대전대학교 한의과대학 사상체질과)
  • Received : 2009.10.24
  • Accepted : 2009.12.02
  • Published : 2009.12.31
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Abstract

1. Objectives: Yeolda-Hanso tang (YH) has long been used as traditional herbal formula in Korea as various diseases. Now we modified Yeolda-Hanso tang (YH) for neurodegenerative diseases treatment and named New-Yeolda-Hanso tang (NYH). We investigated neuroprotective effects of NYH on NGF-differentiated PC12 cells cytotoxicity induced by $\beta$-Amyloid peptide (A$\beta$25-35) and evaluated the ability of NYH to prevent and treat for neurodegenerative diseases via autophagy enhancement. 2. Methods and Results: 1) Protective effect of NYH on PC12 cells cytotoxity induced by A$\beta$25-35. PC12 cells survival was measured by MTT and lactate dehydrogenase (LDH) assay. $20{\mu}M$ $\beta$-Amyloid peptide (A$\beta$25-35) induced cytotoxicity on NGF-differentiated PC12 cells. NYH attenuated the cytotoxic effects of A$\beta$25-35 in a dose-dependent manner. 2) Pharmacological induction of Autophagy by NYH in PC12 cells Autophagy induction and activation was measured by immunoblot assay. Marker of autophagy, LC3 II expression and the ratio of LC3-II/I was slightly increased in the protein treated with YH, and significantly augmented in the protein treated with NYH. NYH-induced increase of LC3-II protein level was inhibited by 3MA. 3) Induction of Autophagy by NYH on A$\beta$25-35-induced injury in PC12 cells In MTT assay, $100{\mu}g/ml$ re-treated NYH attenuated $20{\mu}M$ A$\beta$25-35-induced cytotoxicity in PC12 cells. Protection effect of NYH was blocked by autophagy inhibitor 3MA. In immunoblot assay, $1200{\mu}g/ml$ pre-treated NYH activated autophagy in $20{\mu}M$ A$\beta$25-35-induced cytotoxicity in PC12 cells. The observed effect was partially blocked by 3MA. 3. Conclusions: All the results indicated that NYH possesses neuroprotective potential partially mediated by autophagy enhancement and NYH may be considered to be a promising new herbal formula to prevent and treat for neurodegenerative diseases including Alzheimer's disease (AD).

Keywords

Acknowledgement

Supported by : 한국과학기술원

References

  1. 김지혁 외. 동의 정신의학. 현대의학서적사, 서울, 1992:256-271, 327-330.
  2. 이가옥. 노인생활실태분석및정책과제. 한국보건사회연구원, 1994:114-132.
  3. Butler R.N. geriatric psychiatry. In Kalplan HI, Sadock BJ, eds. Comprehensive testbook of psychiatry V. Baltimore Williams and Wilkins, 1989:2014-2019.
  4. Price D.L, Sisodia S.S. Mutant genes in familial Alzheimer's diseases and transgenic models. Annu. Rev. Neurosci. 1998;21:479-505. https://doi.org/10.1146/annurev.neuro.21.1.479
  5. Selkoe D.J. Alzheimer's diseases: Mechanistic understanding predics novel therapies. Ann. Intern. Med. 2004;140:627-638. https://doi.org/10.7326/0003-4819-140-8-200404200-00010
  6. Lansbury P.T, Lashuel H.A. A century-ole debate on protein aggregation and neurodegeneration enters the clinic. Nature. 2006;443:774-779. https://doi.org/10.1038/nature05290
  7. Olanow C.W, McNaught K.S. Ubiquitin-proteasome system and Parkinson's disease. Mov. Disord. 2006;21:1806-1823. https://doi.org/10.1002/mds.21013
  8. Bandhyopadhyay U, Cuervo AM. Chaperone-mediated autophagy in aging and neurodegeneration: lessons from alpha-synuclein. Exp Gerontol. 2007;42:120-8. https://doi.org/10.1016/j.exger.2006.05.019
  9. Martinez-Vicente M. Cuervo A.M, Autophagy and neurodegeneration: when the cleaning crew goes on strike. Lancet Neurol. 2007;6:352-361. https://doi.org/10.1016/S1474-4422(07)70076-5
  10. Ravikumar B, Duden R, Rubinsztein D.C. Aggregateprone proteins with polyglutamine and polyalanine expansions are degraded by autophagy. Hum Mol Genet. 2002;11:1107-1117. https://doi.org/10.1093/hmg/11.9.1107
  11. 전국한의과대학사상의학교실. 四象醫學. 서울, 집문당, 2004:419-421.
  12. 송일병. 사상의학적중풍관리의임상적연구. 四象體質醫學會紙. 1996;8(2):117-130.
  13. 박수정외6명. 熱多寒少湯이低酸素性大膈神經細胞손상에 미치는 영향. 四象體質醫學會紙. 2003;15(1):72-89.
  14. 이재홍박혜선, 김경요, 고기덕, 김일환. 熱多寒少湯煎湯液이 Hydrogen Peroxide에 의해 損傷된 培養海馬神經細胞에미치는영향. 四象體質醫學會紙. 2002;14(1):79-89.
  15. 최지숙, 김경요, 김형민, 주종천. 人間腦星狀細胞에서熱多寒少湯에의한細胞活性物質生成調節에 關한 硏究. 四象體質醫學會紙. 2001;13(1):61-69.
  16. 이성률, 강형원, 김상태, 류영수. 遠志와石菖蒲混合抽出液의pCT105로유도된신경세포암세포주에 대한 항치매 효과. 東醫生理病理學會紙. 2003;17(4):1037-1049.
  17. 임준모, 이인구,윤종민, 이인, 문병순. 산화적손상에 의한Neuro 2A치매모델에서石菖蒲遠志散의방어효과. 大韓韓醫學會紙. 2005;26(1):161 -173.
  18. 강미선, 장규태, 김장현. 일시적국소뇌허혈흰쥐 모델에서遠志石菖蒲散의 뇌손상및 인지기능 보호효과.東醫生理病理學會紙. 2004;18(6):1777-1783.
  19. 원지상. 東醫四象新編. 정담, 서울 2002.
  20. Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 2000;19:5720-5728. https://doi.org/10.1093/emboj/19.21.5720
  21. Whitehouse PJ, Price DL, Stuble RG, Clark AW, Coyle JT, Delon MR. Alzheimer's disease and senile dementia ; loss of neurona in the basal forebrain. Science. 1982;215(4537):1237-1239. https://doi.org/10.1126/science.7058341
  22. Gridley KE, Green PS, Simpkin JW. Low concentration of estradiol reduce beta-amyloid25-35-induced toxicity, lipid peroxidation and glucose utilization in human SK-N-SH neuroblastoma cells. Brain Res. 1997;778(1):158-165. https://doi.org/10.1016/S0006-8993(97)01056-1
  23. Soto C, Estrada L.D. Protein misfolding and neurodegeneration. Arch Neurol. 2008;65:184-189. https://doi.org/10.1001/archneurol.2007.56
  24. Ciechanover A. Proteolysis: from the lysosome to ubiquitin and the proteasome. Nat Rev. 2005;6:79-86. https://doi.org/10.1038/nrm1552
  25. Dice J.F. Peptide sequences that target cytosolic proteins for lysosomal proteolysis. Trends in Biological Sciences. 1990;15:305-309. https://doi.org/10.1016/0968-0004(90)90019-8
  26. Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 2000;19:5720-5728. https://doi.org/10.1093/emboj/19.21.5720
  27. Seglen P.O, Gordon P.B. 3-methyladenine; specific inhibitor of autophagic/lysosomal protein degradation in isolated rat hepatocytes. Proceeding of the a National Academy of Sciences of U.S.A. 1982;79:1889-1892. https://doi.org/10.1073/pnas.79.6.1889