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The Journal of Internal Korean Medicine > Volume 34(2); 2013 > Article
The Journal of Internal Korean Medicine 2013;34(2): 178-191.
CoCl2로 유도된 C6 신경교세포의 사멸에 대한 抑肝散의 보호 효과
조문영1, 신용진1, 하예진2, 우찬1, 김태정1, 유주연1, 최용석1, 최정훈3, 신선호1
1원광대학교 한의과대학 내과학교실
2한국보건산업진흥원 한의약연구지원팀
3함소아 한의원
Protective Effects of Ukgan-san in CoCl2-induced Cell Death of C6 Glial Cells
Mun-young Cho1, Yong-jeen Shin1, Ye-jin Ha2, Chan Woo1, Ta-jung Kim1, Ju-yeon You1, Yong-seok Choi1, Jung-hoon Choi3, Sun-ho Shin1
1Dept. of Internal Medicine, College of Oriental Medicine, Won-Kwang University
2Traditional korean medicine research support team, Korea health industry development institute
3Hamsoa Oriental Medical Clinic
Correspondence  Sun-ho Shin ,Tel: 063-270-1013, Fax: 063-270-1594, Email: shshin@wonkwang.ac.kr
  Published online: June 30, 2013.
ABSTRACT
Objectives:
In this study, we made an effort to investigate the protective mechanism of Ukgan-san (UGS) extracts on hypoxia-induced C6 glial cell death.

Methods :
The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MMT) assay and cell morphological changes were analysed with microscope after staining with crystal violet (CV). Reactive oxygen species (ROS) formation was assessed by flow cytometer after staining with 2'7'-dichlorofluorescein diacetate (DCF-DA). We also analyzed expression of hypoxia-inducible factor-1 alpha (HIF-1α) and p53, processing of procaspase-3 and procyclic acidic repetitive protein (PARP) by western blot method.

Results:
We estimated the elevated cell viability by UGS extract on CoCl2-induced C6 glial cells. UGS attenuated CoCl2-induced ROS formation in C6 glial cells and also showed a protective activity compared to antioxidants and exhibited abrogation of LDH-released by CoCl2. UGS suppressed the typical apoptotic cell death markers, caspase-3 and PARP activation. UGS inhibited CoCl2-induced HIF-1α expression which is known as a major regulator for hypoxia-induced cell death, and suppressed p53 expression.

Conclusions:
These results suggest that UGS extract contains protective constituents for hypoxia-induced C6 glial cell death.
Key words: CoCl2, C6 glial cell, protective effect, gan-san (Yigan-san)
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