• 대한한방내과학회지
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• 제22권1호
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• pp.87-93
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• 2001

A human hepatoma cell line, Hep G2 cells, is reliable for the study of alcohol-induced hepatotoxicity. The aim of this study is to determine the relationship between TNF-${\alpha}$, IL-$1{\alpha}$ production and EtOH-induced cytotoxicity on Hep G2 cells. The cells were incubated with EtOH in the presence of Artemisia Capillaris Herba(AC) for 24 hours and in the absence of AC for 48 hours. Cytoviability and cytokines release were analyzed by MTT assay and enzyme linked immunosorbent assay (ELISA), respectively. After 24 hours of EtOH exposure, the cytoviability had markedly decreased, and the release of cytokines had increased. The increased amount of cytokines contributed to EtOH-induced cytotoxicity. Anti-TNF-${\alpha}$ and IL-$1{\alpha}$ antibodies almost abolished it. Interestingly, EtOH-induced cytotoxicity and cytokines production were inhibited by AC. Moreover, when AC was used in combination with antibodies, there was a marked inhibition of EtOH-induced cytotoxicity. These results suggest that EtOH-induced cytotoxicity may regulate, by various factors, and AC may prevent the cytotoxicity through partial inhibition of the $TNF-{\alpha}$ and IL-$1{\alpha}$ secretion.

• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.43-50
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• 2002

• Advances in Traditional Medicine
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• 제1권1호
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• pp.89-96
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• 2000

A human hepatoma cell line, Hep G2 cells are a reliable for the study of alcohol-induced hepatotoxicity. In this study, the author investigated the effect of an aqueous extract of Asparagus $cochinchinensis_{MERRIL}$ (Liliaceae) roots (ACAE) on ethanol (EtOH)-induced cytotoxicity in Hep G2 cells. ACAE dose-dependently inhibited the EtOH-induced tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ secretion. ACAE also inhibited the EtOH and $TNF-{\alpha}-induced$ cytotoxicity. Furthermore, the author found that ACAE inhibited the $TNF-{\alpha}-induced$ apoptosis of Hep G2 cells. These results suggest that ACAE may prevent the EtOH-induced cytotoxicity through inhibition of the apoptosis of Hep G2 cells.

• 동의생리병리학회지
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• 제25권4호
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• pp.663-668
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• 2011

Galgeun-tang (GGT) has been a great source for treating cold diseases in the folk medicine recipe. Carbon tetrachloride ($CCl_4$) is one type of hepatotoxin that can eventually cause liver injury. During the experiment, we first studied the protective effects of GGT against $CC_4$-induced hepatotoxicity. GGT was pretreated for 3 h, and 1% $CCl_4$ was added to mouse primary liver cells. After 4 h, ROS generation and expression of antioxidant enzymes (catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx)) were analyzed by FACS and real time PCR. Also, the activities of ALT and LDH were measured using cultured medium. The hepatic levels of TNF-alpha and iNOS, which are related to inflammation and stress response gene, HSP72 and HO-1 were analyzed by PCR or real time PCR. Liver tissues were analyzed by HE stain. From the observation, we discovered that GGT treatment protects $CCl_4$-induced hepatotoxicity, and that GGT pretreatment decreases ROS generation, TNF-alpha and iNOS expression. However, gene expression of CAT, SOD, GPx, HSP72 and HO-1 were increased by GGT. These results lead to the conclusion that GGT has protective effects against $CCl_4$-induced hepatotoxicity.

• 한국식품영양과학회지
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• 제39권3호
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• pp.343-349
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• 2010

녹차 추출물이 에탄올에 의한 조기 간세포에 미치는 영향을 관찰하기 위해 흰쥐에 에탄올과 녹차 추출물을 투여하고 1시간 및 3시간 후 흰쥐의 간 글루타치온과 TNF-$\alpha$량, 간항산화효소 활성도, 지질과산화물량 및 혈청 AST 및 ALT 활성도를 측정하였다. 또한 에탄올에 의한 만성 간 손상에 미치는 영향을 관찰하기 위해서 에탄올과 녹차 추출물을 2일 간격 투여 20일 후 흰쥐의 간 글루타치온과 TNF-$\alpha$량, 간 항산화 효소활성도, 지질과산화물량 및 혈청 AST 및 ALT 활성도를 측정하였다. 간 글루타치온량은 대조군에 비하여 에탄올 투여군에서 1시간, 3시간 및 20일 경과 후에 모두 감소되었고, 에탄올과 녹차추출물 혼합투여로 간 글루타치온량이 에탄올 투여군에 비하여 증가되어 녹차추출물이 에탄올에 의한 간 글루타치온량 감소를 억제시킴을 알 수 있다. 간 TNF-$\alpha$량은 에탄올 투여 1시간, 3시간 및 20일경과 후 증가되었고, 녹차추출물과 에탄올 혼합투여군은 에탄올 투여군에 비하여 1시간, 3시간 및 20일 경과 후 모두 감소되어 녹차추출물 투여로 에탄올 투여에 의한 간조직의 TNF-$\alpha$량 증가가 억제됨을 알 수 있다. 에탄올 투여군의 간조직 TBARS량을 측정한 결과 에탄올을 투여한 후 1시간 경과 및 3시간 경과 후 대조군과 유의한 차이가 없었다. 에탄올 투여군의 간 조직 항산화효소(SOD, catalase, GPx) 활성도를 측정한 결과 에탄올을 투여한 후 1시간 경과 및 3시간 경과 후 대조군과 유의한 차이가 없었다. 에탄올 투여군의 혈청 ALT와 AST 활성도를 측정한 결과 에탄올을 투여한 후 1시간 경과 및 3시간 경과 후 대조군과 유의한 차이가 없었다. 이상의 실험 결과 녹차 추출물은 간에서 에탄올에 의해서 조기에 일어나는 글루타치온량 감소 및 TNF-$\alpha$량 증가를 억제시키는 작용이 있는 것을 알 수 있는데, 녹차 추출물의 이러한 효과는 녹차 추출물이 에탄올에 의한 간 손상을 억제하는데 중요한 작용을 할 수 있을 것으로 기대된다. 녹차 추출물에 함유된 에탄올에 의한 간 손상 방지 물질의 검색과 글루타치온 및 TNF-$\alpha$에 대한 작용 기전 및 특성에 대한 연구가 계속된다면 우수한 간 손상 방지작용을 가진 물질이 개발될 수 있을 것으로 추측된다.

• The Korean Journal of Physiology and Pharmacology
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• 제20권1호
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• pp.15-23
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• 2016

Acetaminophen (APAP) overdose is one of the most common causes of acute liver failure. The study aimed to investigate the protective effect of carnosic acid (CA) on APAP-induced acute hepatotoxicity and its underlying mechanism in mice. To induce hepatotoxicity, APAP solution (400 mg/kg) was administered into mice by intraperitoneal injection. Histological analysis revealed that CA treatment significantly ameliorated APAP-induced hepatic necrosis. The levels of both alanine aminotransferase (ALT) and aspartate transaminase (AST) in serum were reduced by CA treatment. Moreover, CA treatment significantly inhibited APAP-induced hepatocytes necrosis and lactate dehydrogenase (LDH) releasing. Western blot analysis showed that CA abrogated APAP-induced cleaved caspase-3, Bax and phosphorylated JNK protein expression. Further results showed that CA treatment markedly inhibited APAP-induced pro-inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, IL-6 and MCP-1 mRNA expression and the levels of phosphorylated $I{\kappa}B{\alpha}$ and p65 protein in the liver. In addition, CA treatment reduced APAP- induced hepatic malondialdehyde (MDA) contents and reactive oxygen species (ROS) accumulation. Conversely, hepatic glutathione (GSH) level was increased by administration of CA in APAP-treated mice. Mechanistically, CA facilitated Nrf2 translocation into nuclear through blocking the interaction between Nrf2 and Keap1, which, in turn, upregulated anti-oxidant genes mRNA expression. Taken together, our results indicate that CA facilitates Nrf2 nuclear translocation, causing induction of Nrf2-dependent genes, which contributes to protection from acetaminophen hepatotoxicity.

• 약학회지
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• 제54권1호
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• pp.55-61
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• 2010

Gardenia jasminoides is one of the most widely used herbal preparations for the treatment of liver disorders. This study evaluated the potential beneficial effect of G. jasminoides in a mouse model of carbon tetrachloride ($CCl_4$)-induced liver injury. The mice were treated intraperitoneally with $CCl_4$ (10 ${\mu}l$/kg). They received G. jasminoides (30, 100, 300 mg/kg) 48 h, 24 h and 2 h before and 6 h after administering $CCl_4$. The serum activities of aminotransferase and the hepatic level of malondialdehyde were significantly higher 24 h after the $CCl_4$ treatment, while the concentration of reduced glutathione was lower. These changes were attenuated by G. jasminoides. $CCl_4$ increased the level of circulating tumor necrosis factor-$\alpha$ (TNF-$\alpha$) markedly, which was reduced by G. jasminoides. The levels of hepatic inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expression were markedly higher after the $CCl_4$ treatment. G. jasminoides diminished these alterations. $CCl_4$ increased the level of TNF-$\alpha$, iNOS and COX-2 mRNA expressions, and these increases were attenuated by G. jasminoides. These results suggest that G. jasminoides alleviates $CCl_4$-induced liver injury, and this protection is likely due to the reduced oxidative stress and the downregulation of proinflammatory mediators.

• 약학회지
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• 제43권2호
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• pp.198-207
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• 1999

The aim of the present investigation was to examine whether YH439, a hepatoprotective agent, exerts protective effect against hepatotoxicity and reduces the production of cytokines and NO in lipopolysaccharide (LPS)-primed rats with carbon tetrachloride ($CCl_4$). Administration of LPS following a single dose of CCl4 injection resulted in remarkable elevations of the serum $TNF{\alpha},{\;}IL-l{\beta$ and IL-6 level. The serum NO level was moderately elevated and severe liver damage was evidenced by increases in serum alanine aminotransferase (ALT) and sorbitol dehydrogenase (SDH) activities. YH439 decreased the levels of TNF, $IL-l{\beta}$, IL-6, ALT, SDH as well as NO in the serum elevated by CCl4+LPS in a dose-dependent manner. Inducible nitric oxide synthase (iNOS) level was decreased in the liver of rats treated with YH439. The increased iNOS activity induced by LPS and $interferon-{\gamma}$ was significantly decreased in RAW 264.7 cells by YH439 treatment. YH439 increased the GSH level decreased by $CCl_4+LPS$ and suppressed the ratio of GSSG/GSH. The reduction of hepatotoxicity by YH439 may associated with the decrease in the production of cytokines as well as suppression of iNOS protein in conjunction with an increase in the GSH level.

• IMMUNE NETWORK
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• 제1권3호
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• pp.213-220
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• 2001

Background: A previous study has shown that Euonymus alatus (EA) has an antidotic activities against inflammation, suggesting possibility that EA can exert this beneficial effects to liver injury by an initial protection against drug-induced hepatocyte demage. The present study was undertaken to evaluate the protective effect of EA-extract on experimentally induced hepatitis in ICR mice and to investigate some mechanisms responsible for its action. Methods: Water EA extract was used in this experiments. The mice received i.p. a dose of 700 mg/kg galactosamine (GalN) together with $5{\mu}g/kg$ of endotoxin (LPS), or received i.v. 12 mg/kg of concanavalin A (Con A). EA (4 mg/mouse) was administrated on day -2, -1 and 0 before induction of liver injury. Liver injury was assessed by measurement of serum alanin amino-transferase (SGPT) levels on 9 hr after GaIN.LPS, or 8 hr after con A administration. Results: Treatment with either GaIN or LPS alone did not cause hepatitis. However, simultaneous administration of GalN and LPS to mice resulted in LPS-dose dependent fulminant hepatitis. GaLN/LPS-induced liver injury was reduced when mice were given EA for 3 days before induction. This preventive effect of Ea was more prominent when EA was given by intraperitoneal route rather then by oral route. Pretreatment of EA or dexamethasone inhibited significantly $TNF{\alpha}$ production in GalL/LPS-injured mice. However, EA-treatment did not influence $TNF{\alpha}$-induced hepatitis in GalN-sensitized mice, suggesting that $TNF{\alpha}$ is likely to act as one of final mediators of endotoxin action and the protective effect of EA might be manifested chiefly by inhibition of endotoxin-induced $TNF{\alpha}$ production, not by blocking the $TNF{\alpha}$-action. Injection of Con A into mice evoked remarkable liver injury in a dose dependent fashion. This liver damage was reduced by EA-pretreatment. Dexamethasone significantly reduced both GalL/LPS-induced and Con A-induced liver damages, showing synergism with EA. However, indomethacin reduced only GalN/ LPS-induced hepatitis, not for Con A-induced hepatitis. Conclusion: These results led to the conclusion that EA may be able to contribute at least in part to prevent the drug-induced hepatotoxicity, and that its anti-hepatitis effects might be manifested directly by modulation of endogenous mediators, such as leukotriese D4, $TNF{\alpha}$ and free radical, and indirectly by regulation of immune mediated responses. Also these results suggested that EA could be developed as a potential antidotic agent.

• Toxicological Research
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• 제20권3호
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• pp.233-239
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• 2004

The present study was designed to determine the impact of mercury on endotoxin-induced inflammatory cytokine expression and corresponding signal transduction in mouse kidney. Male BALB/c mice were exposed continuously to 0, 0.3, 1.5, 7.5, or 37.5 ppm of mercury in drink-ing water for 14 days and at the end of the treatment period, lipopolysaccharide (LPS, 0.5 mg/kg) was injected intraperitoneally 2 h prior to euthanasia. The doses of mercury and LPS did not cause hepatotoxicity or renal toxicity as indicated by unaltered plasma alanine aminotransferase and aspartate aminotransferase levels, and terminal UTP nucleotide end-labeling assay from kidney, respectively. Mercury decreased kidney glutathione (GSH) and with LPS, it additively decreased GSH. Mercury activated p38 mitogen-activated protein kinase (MAPK) and additively increased LPS-induced p38 MAPK phosphorylation. In contrast, mercury inhibited LPS-induced activation of extra-cellular signal-regulated kinase (ERK) but had no effect alone. Mercury increased the gene expression of tumor necrosis factor $\alpha$ (TN F$\alpha$) and potentiated LPS-induced TNF$\alpha$ expression. Mercury did not affect LPS-induced interleukin-1$\beta$ (IL-1$\beta$) expression but decreased LPS-induced IL-6 expression. These results suggest that low levels of mercury might augment LPS-induced TNF$\alpha$ expression by altering GSH and p38 MAPK. Mercury modulates LPS-induced p38 and ERK activation, and downstream TNF$\alpha$ and IL-6 expression in kidney, respectively.