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CoCl2로 유도된 C6 신경교세포의 사멸에 대한 抑肝散의 보호 효과

Protective Effects of Ukgan-san in CoCl2-induced Cell Death of C6 Glial Cells

유주연|최용석|신용진|김태정|최정훈|신선호|하예진|우찬|조문영
대한한방내과학회 (The Society Of Korean Oriental Internal Medicine)|2013.06|대한한방내과학회지 (The Journal of Korean Oriental Internal Medicine)|제34권 제2호|pp. 178-191

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참고문헌

키워드

CoCl2protective effectC6 glial cellUkgan-san (Yigan-san)
주제분류한의학

초록

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.

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.

참고문헌 (34)

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