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KMID : 0363320120330030272
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Journal of Korean Oriental Internal Medicine
2012 Volume.33 No. 3 p.272 ~ p.283
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Protective Effect of Jinmu-tang on -induced Cell Death in C6 Glial Cells
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Choi Jung-Hoon
Shin Yong-Jeen
Ha Ye-Jin
Cho Mun-Young
You Ju-Yeon
Lee Soong-In
Shin Sun-Ho
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Abstract
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Objectives: The purpose of this study was to investigate the mechanism of protective effect of Jinmu-tang (JMT, Zhenwu-tang) extract on -induced cell death in C6 glial cells.
Methods: Cultured C6 glial cells of white mice were pretreated with JMT extract and exposed to 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- (NF-) by western blot method. Tumor necrosis factor- (TNF-) secretion was analyzed using Quantikine kit.
Results: We determined the elevated cell viability by JMT extract on -induced C6 glial cell death. ROS formation, DNA fragmentation, phosphorylation, NF- activation, and secretion of TNF- induced by 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 -induced C6 glial cell death in various pathways.
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KEYWORD
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C6 glial cell, Protective effect, Jinmu-tang (Zhenwu-tang)
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