• The Journal of Korean Medicine
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• v.24 no.1
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• pp.190-201
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• 2003

Object : This study was designed to investigate the effects of Chungganhaeju-tang (Qingganjiejiu-tang) on cytotoxicity, growth inhibition, apoptosis and expression of cytokine in damaged HepG2 cells. Method : Toxicity on HepG2 cell induced by ethanol and acetaldehyde was measured for viability, cell growth, DNA replication and generation of apoptosis and cytokine. The recovery of the cell activity by Chungganhaeju-tang was estimated for the measured parameters using PCR with different cycle numbers, DNA gel-electrophoresis, and densitometric analysis, Results : Chungganhaeju-tang improves the recovery of HepG2 cells damaged by ethanol or acetaldehyde. The suppressed DNA synthesis of the cell damaged by ethanol or acetaldehyde is improved by Chungganhaeju-tang. A liver-protection effect was shown by the reduction of apoptosis and $TNF-{\alpha},{\;}IL-1{\beta}$ expressions that are induced by ethanol or acetaldehyde. Conclusion : The result indicates that Chungganhaeju-tang reduces toxicity induced by ethanol or acetaldehyde and recovers damaged liver function.

• The Journal of Internal Korean Medicine
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• v.25 no.1
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• pp.81-91
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• 2004

Objectives : The aim of this study is to investigate the inhibitory effect of Chungganhaeju-Tang on alcohol induced human hepatic cell apoptosis by synthesis of glutathione. Methods : The amount of glutathione in HepG2 cell was measured with colorimetric glutathione assay kit and glutathione-conjugated CDNB(1-chloro-2,4-dinitrobenzene) at $37^{\circ}C$ and then measured by spectrometry to assess the activity of glutathione S-transferase. Results : The synthesis of glutathione and the activity of glutathione S-transferase in HepG2 cell were promoted by Chungganhaeju-Tang and increased in dose/time-dependent manner. Chungganhaeju-Tang inhibited apoptosis induced by ethanol and acetaldehyde dependent to treatment dosage. In Buthione sulfoximine, a glutathione synthesis inhibitor, treated case, the synthesis of glutathione was inhibited and in Chungganhaeju-Tang treated case, the synthesis of glutathione is promoted with or without Buthione sulfoximine. The present findings suggest that Chungganhaeju-Tang inhibits alcohol induced apoptosis by synthesis of glutathione in HepG2 cell. Conclusions : The result indicates that Chungganhaeju-Tang protects human hepatic cell by glutathione synthesis and made the liver recover from alcohol induced damage.

• The Journal of Internal Korean Medicine
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• v.28 no.1
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• pp.68-79
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• 2007

Objectives : The purpose of our study was to investigate the effects of Chungganhaeju-tang (Qingganjiejiu-tang) on alcoholic liver damage by applying proteomics. Materials and Methods : Sprague-Dawley rats were used in this experiment; the rats were divided into a control group, alcohol group and Chungganhaeju-tang + alcohol group. Ethanol was orally administered twice a day for 4 weeks to the alcohol group. Water without ethanol was administered twice a day for 4 weeks to the control group. Ethanol + Chungganhaeju-tang extract was orally administered twice a day for 4 weeks to the Chungganhaeju-tang + alcohol group. The livers of each group were processed and we investigated histology, OxyBlot, 2-dimensional electrophoresis, and western blot of liver of each group. Results : In the histological findings of the liver, the alcohol group showed portal fibrosis with a few septa or without septa. The Chungganhaeju-tang + alcohol group showed no fibrosis or portalfibrosis without septa. In the OxyBlot finding, Chungganhaeju-tangprevented liver damage by oxidation. In the 2-dimensional electrophoresis finding, formiminotransferase cyclodeaminase (FTCD), glucose regulated protein, 58 kDa (GRP58K), aryl sulfotransferase, sulfotransferase family 1A, member 2, similar to acyl-coenzyme A oxidase-like, and catalase were changed. Conclusion : Chungganhaeju-tangexerts an inhibitory effect against the fibrosis and oxidation induced by alcohol in rat liver cell, and some proteins induced by alcohol were changed by Chungganhaeju-tang.

• The Journal of Internal Korean Medicine
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• v.25 no.1
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• pp.59-70
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• 2004

Objectives : Previous studies showed that treatment with Chungganhaeju-tang prevents hepatic inflammation and apoptosis in alcoholic liver disease. The purpose of our study is to determine if any relations exsists between the transcription factor $NF{\kappa}B$, an orchestrating expression of a large number of genes and inhibitory effects of Chungganhaeju-tang on ethanol induced apoptosis. Materials and Methods : To assess the role of $NF{\kappa}B$, we blocked NFkB activation by delivering to the kupffer cells $I{\kappa}B{\Delta}N$, a dominant negative $NF{\kappa}B$ inhibitor, and investigated if Chungganhaeju-tang still prevented apoptosis. Results : When $NF{\kappa}B$ activation was blocked, there was no inhibitory effect of Chungganhaeju-tang on ethanol induced apoptosis of kupffer cells. Conclusion : This result suggests that Chungganhaeju-tang protects the liver from ethanol induced apoptosis by activating the $NF{\kappa}B$ that plays a key role in porotecting mechanism and reducing inflammatory cytokine gene expression.

• The Journal of Korean Medicine
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• v.22 no.4
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• pp.107-113
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• 2001

Objectives : The aim of this study was to investigate effects of Chungganhaeju-tang on alcoholic fatty liver. Methods : Thirty patients with alcoholic fatty liver were included in this study. Chungganhaeju-tang was administered to subjects for 1 month. The changes of clinical symptoms and chemical liver function tests were evaluated before and after treatment. Results : Chungganhaeju-tang has effects on the improvement of clinical symptoms such as fatigue, anorexia, dyspepsia, nausea, and RUQ (Right Upper Quadrant) pain. On the chemical liver function test, serum AST, ALT, GGT, and TG level significantly decreased (P<0.05, by paired t-test). Conclusions : These results suggest that Chungganhaeju-tang has significant effects on the improvement of clinical symptoms and chemical liver function test with alcoholic fatty liver.

• The Journal of Korean Medicine
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• v.21 no.1
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• pp.68-76
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• 2000

Objectives: This experiment was conducted to verify the effects of Chungganhaeju-tang(Qingganjiejiu-tang) on the alcohol metabolism and liver functions, by measuring the activity levels of ADH and ALDH, as well as glucose, triglyceride, and BUN. Damage of the liver cells caused by alcohol was determined through the examination of serum levels of AST, ALT, ALP, LDH, and uric acid. Methods: Sprague-Dawley rats were used in this experiment and the rats were divided into control and experiment groups. Chungganhaeju-tang(Qingganjiejiu-tang) extract was orally administered in the experiment group for three weeks. Each group was further classified into two sub-groups, and control group's blood was taken without oral ingestion of alcohol, while the experiment group' s blood was withdrawn after ingestion of alcohol. Evaluation of damage level was done considering the presence of extract and alcohol. Results: In this experiment, Chungganhaeju-tang(Qingganjiejiu-tang) significantly suppressed the activity of ADH which is a precursor enzyme of acetaldehyde, but didn't cause significant changes in the activity of ALDH which is a catabolic enzyme. Decreased glucose level due to alcohol consumption was recovered back to the normal level and increased levels of triglyceride, BUN, AST, ALT, ALP, LDH, and uric acid were significantly reduced. Conclusions: These experiment results suggest that Chungganhaeju-tang(Qingganjiejiu-tang) inhibits the formation of acetaldehyde in the metabolism of alcohol, and affects the recovery of weakened liver functions due to alcohol.

• The Journal of Internal Korean Medicine
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• v.27 no.1
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• pp.27-39
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• 2006

Objectives : Induction of CYP2E1 by ethanol is believed to be one of the major mechanism by which ethanol generate a state of oxidative stress. Previous studies showed that treatment with Chungganhaeju-tang prevents hepatic inflammation and apoptosis in alcoholic liver disease. The purpose of our study is to determine if Chungganhaeju-tang can also protect against alcohol-induced cytotoxicity in CYP2E1-transfected HepG2 cells. Materials and Methods : CYP2E1-transfected HepG2 cells and control vector-transfected HepG2 cells were exposed for isx hours to Chungganhaeju-tang, and then 50 mM of ethanol was added and left for two days. Results : Ethanol significantly decreased cell viability in CYP2E1-transfected HepG2 cells and increased apoptosis. These alterations were attenuated by Chungganhaeju-tang. This was accompanied by an improvement of NF-${\kappa}B$ and Akt activation. Conclusion : These results suggest that Chungganhaeju-tang exerts inhibitory effect against the cytotoxicity induced by alcohol in CYP2E1-transfected HepG2 cells, and that this is a protective action due, at least in part, to an activation of NF-${\kappa}B$ that plays a key role in the protection mechanism, and in reducing hepatotoxic cytokine gene expression.

• The Journal of Internal Korean Medicine
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• v.24 no.1
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• pp.123-133
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• 2003

Objectives : This study was designed to investigate the effects of Chungganhaeju-tang on expression of alcohol metabolizing enzymes, cell viability and alcohol-induced apoptosis. Materials and Methods : For this study, the human hepatoma cell line HepG2 was used. HepG2 cells were treated with ethanol-or acetaldehyde, chungganhaeju-tang, anti-Fas neutralizing antibody and were investigated by using quantitative RT-PCR, MTT and Trypan blue exclusion assays. Results : The results are summarized as follows: 1. Quantitative RT-PCR analysis demonstrated that ethanol-or acetaldehyde-mediated increase of ALDH gene expression was not affected by Chungganhaeju-tang treatment. 2, Ethanol-or acetaldehyde-induced apoptosis was remarkably inhibited by Chungganhaeju-tang in a dose-dependent manner. 3, Ethanol-or acetaldehyde-induced apoptosis was significantly blocked by anti-FasL neutralizing antibody, suggesting apoptosis induced by alcohol might be mediated by FasL/Fas signaling pathway. Conclusions : Taken all together, these results indicate that the FasL/Fas signaling plays a critical role in alcohol-induced apoptosis and Chungganhaeju-tang increases viability of liver cells by suppression of the FasL/Fas-mediated apoptosis-signaling pathway.

• The Journal of Internal Korean Medicine
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• v.26 no.1
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• pp.93-106
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• 2005

Objectives : The aim of this study is to characterize the effect of Chungganhaeju-tang on $TGF-{\beta}l$-induced hepatic fibrosis. Materials and Methods : mRNA and protein expression levels of $TGF-{\beta}l$ in Chungganhaeju-tang treated HepG2 cells were compared to untreated cells using quantitative RT-PCR and ELISA assay, respectively. mRNA expression levels of the $TGF-{\beta}l$ signaling pathway genes $(T{\beta}R-I,\;T{\beta}R-II,\;Smad2,\;Smad3,\;Smad4,\;and\;PAI-1)$ and fibrosis-associated genes (CTGF, fibronectin, and collagen type $l{\alpha}$) were evaluated by quantitative RT-PCR. The effect of Chungganhaeju-tang on cell proliferation of T3891 human fibroblast was evaluated using $[^3H]Thymidine$ Incorporation Assay. Results : Inhibition of $TGF-{\beta}l$ mRNA expression and protein production was observed with treatment of Chungganhaeju-tang and seen to be dose and time dependent. Whereas $TGF-{\beta}l$-mediated induction of PAI-1 was suppressed with treatment of Chungganhaeju-tang, expression of the $TGF-{\beta}l$ signaling pathway genes such as $T{\beta}R-I$, $T{\beta}R-II$, Smad2, Smad3, and Smad4 was not affected. With treatment of Chungganhaeju-tang, inhibition of $TGF-{\beta}l$-induced cell proliferation of T3891 human fibroblast was observed, as well as abrogation of $TGF-{\beta}l$-mediated transcriptional up-regulation of CTGF, fibronectin, and collagen type $I{\alpha}$. Conclusion : This study strongly suggests that the liver cirrhosis-suppressive activity of Chungganhaeju-tang may be derived at least in part from its inhibitory effect on $TGF-{\beta}l$ functions, such as blockade of $TGF-{\beta}l$ stimulation of fibroblast cell proliferation and fibrosis-related gene expression as well as expression of $TGF-{\beta}l$ itself.

• The Journal of Internal Korean Medicine
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• v.32 no.2
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• pp.188-202
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• 2011

Objectives : Oxidative stress seems to play a major role in mechanisms by which ethanol causes liver injury. Previous studies have shown that treatment with Chungganhaeju-tang (Qingganjiejiu-tang, CGHJT) has protective effects on alcoholic liver disease. The aim of this study was to investigate the effects of Chungganhaeju-tang on oxidative stress. Materials and Methods : In vitro, we evaluated the inhibitory activities of CGHJT on DPPH (1,1-diphenyl-2-picryl-hydrazyl), xanthine oxidase, trypsin, and hyaluronidase, and measured cell viability, and proliferation. In the cell culture model, we measured the activities of superoxide dismutase (SOD), and catalase (CAT) after CGHJT treatment in C34 and E47 cell lines, HepG2 cells transfected with/without the cytochrome P450 2E1 (CYP2E1) gene. In vivo, we measured malondialdehyde levels in the liver tissue and alcohol concentration in the blood. Results : CGHJT showed significant free radical scavenging activity against DPPH and xanthine oxidase in the in vitro study, and increased cell viability, proliferation, and activities of superoxide dismutase, catalase in C34 and in E47 cell lines. CGHJT reduced malondialdehyde levels and blood alcohol concentration in vivo, as well. Conclusions : This study suggests that CGHJT has antioxidant effects on oxidative stress by reducing lipid peroxidation and inhibiting the ethanol induced suppression of antioxidant enzyme activities.