The Journal of Internal Korean Medicine (대한한방내과학회지)

The Society of Internal Korean Medicine (대한한방내과학회)
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Protective Effect of Jinmu-tang on $H_2O_2$-induced Cell Death in C6 Glial Cells

진무탕(眞武湯)이 $H_2O_2$로 유도된 C6 Glial 세포사에 미치는 영향

  • Choi, Jung-Hoon (Hamsoa Oriental Medical Clinic) ;
  • Shin, Yong-Jeen (Dept. of Internal Medicine, College of Oriental Medicine, Won-Kwang University) ;
  • Ha, Ye-Jin (Dept. of Internal Medicine, College of Oriental Medicine, Won-Kwang University) ;
  • Cho, Mun-Young (Dept. of Internal Medicine, College of Oriental Medicine, Won-Kwang University) ;
  • You, Ju-Yeon (Dept. of Internal Medicine, College of Oriental Medicine, Won-Kwang University) ;
  • Lee, Soong-In (Sangsang Oriental Medical Clinic) ;
  • Shin, Sun-Ho (Dept. of Internal Medicine, College of Oriental Medicine, Won-Kwang University)
  • 최정훈 (함소아한의원) ;
  • 신용진 (원광대학교 한의과대학 내과학교실) ;
  • 하예진 (원광대학교 한의과대학 내과학교실) ;
  • 조문영 (원광대학교 한의과대학 내과학교실) ;
  • 유주연 (원광대학교 한의과대학 내과학교실) ;
  • 이숭인 (생생한의원) ;
  • 신선호 (원광대학교 한의과대학 내과학교실)
  • Published : 2012.09.30
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Abstract

Objectives : The purpose of this study was to investigate the mechanism of protective effect of Jinmu-tang (JMT, Zhenwu-tang) extract on $H_2O_2$-induced cell death in C6 glial cells. Methods : Cultured C6 glial cells of white mice were pretreated with JMT extract and exposed to $H_2O_2$ for inducing cell death. We measure the cell viability by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and investigate the cell morphology using a light microscope after crystal violet (CV) staining. Reactive oxygen species (ROS) formation was analyzed using a flow cytometer and a fluorescent microscope after staining with 2'7'-dichlorofluorescein diacetate (DCF-DA). DNA fragmentation was analyzed using a flow cytometer after propidium iodide (PI) staining and nuclei morphology was investigated using a fluorescent microscope after 2-[4-amidinophenyl]-6-indo-lecarbamidine dihydrochloride (DAPI) staining. We analyzed expression of Bax, processing of procaspase-3 and poly (ADP-ribose) polymerase (PARP), and activation of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) by western blot method. Tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) secretion was analyzed using Quantikine kit. Results : We determined the elevated cell viability by JMT extract on $H_2O_2$-induced C6 glial cell death. ROS formation, DNA fragmentation, $I{\kappa}B{\alpha}$ phosphorylation, NF-${\kappa}B$ activation, and secretion of TNF-${\alpha}$ induced by $H_2O_2$ are inhibited by JMT extract pre-treatment. JMT extract inhibits Bax expression, processing of caspase-3 and PARP that are critical biochemical markers of apoptotic cell death. Conclusions : These results suggest that JMT extract has a protective effect on $H_2O_2$-induced C6 glial cell death in various pathways.

Keywords

References

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