• 대한한의학회지
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• 제20권3호
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• pp.94-104
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• 1999

Objectives: Hepatoma is a very serious disease in Korea and worldwide. Hepatitis B virus (HBV) has proved the most significant cause of hepatoma. We carried out this study to investigate the effect of Injinchunggan-tang (Yinchenqinggan -tang) on inhibiting cell proliferation and DNA synthesis in HepG2.2.15 cell lines and on inhibiting phosphorilation of oncogene (MAP kinase) in NIT /3T3-HEx cells. Methods: First we confinned the Hepatitis B virus producing ability of HepG2.2.15 cells. To investigate the anti-cancer effect of Injinchunggan-tang (Yinchenqinggan-tang), we did the NTS/PMS assay, [3H]-thymidine incorporation assay and transfection of pcDNA-X. We also measured the gene expression through western blotting. Results: Injinchunggan-tang (Yinchenqing gan tang) showed the suppressing effect of HepG2.2.l5 increase in the MTS/PMS assay and the inhibiting effect of DNA synthesis of HepG2.2.15 in the [3H] thymidine incorporation assay. Injinchunggan-tang (Yinchenqinggan-tang) also showed the inhibiting phosphorilation effect of MAP kinase in HBV -X genes. Conclusions: From the above Injinchunggan-tang (Yinchenqinggan-tang) is thought to have an anti-cancer effect on the hepatoma from HBV. It is suggested that further studies on this prescription would give us a better medicine with an anti-cancer effect.

• 한국환경성돌연변이발암원학회지
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• 제24권4호
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• pp.171-179
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• 2004

Cytochrome P4503A4(CYP3A4) is the most abundnat CYPs in human liver, comparising approximately 30% of the total liver CYPs contents ans is involbed in the metabolism of more than 60% of currently used therapeutic drugs. The expression of CYP3A4 is induced by a variety of structurally unrelated xonobiotics including the antibiotic rifampicin and endogenous hormones, and might be mediated through steroid and xenobiotic receptor(SXR) system. The molecular mechanisms underlying regulation of CYP3A4 gene expression hae not been understood. In order to gain the insight of the molecular mechanism of CYP3A4 gene expression, study has been undertaken to investigate if the histone deacelylation is involved in the regulation of CYP3A4 gene expression by proximal promoter or not. Also SXR was investigated to see if they were involved in the regulation of CYP3A4 proximal promoter activity. HepG2 or Hena-I cells were transfected with a plasmid containing~1kb of the CYP3A4 proximal promoter region (-863 to +64bp) cloned in front of a reporter gene, luciferase, in the presence or absence of SXR or hER. Transfected cells were treated with CYP3A4 inducers such as rifampicin, PCN and RU 486, or with estradiol, in order to exmine to regulation of CYP3A4 gene expression in the presence or absence of trichostatin A (TSA). In HepG2 cells, CYP3A4 inducers and estradiol increased significantly the luciferase activity by CYP3A4 proximal promoter, only when TSA was co-treated after SXR cotransfection. In the case of Hepa-I cells CYP3A4 inducers and estradiol incressed modestly the luciferase activity when TSA was co-treated, but this increment was not enhanced by SXR cotransfection in contrast to HepG2 cells. Taken together, these results indicated that the inhibition of histone deacetylation was required to SXR-mediated increase in CYP3A4 proximal promoter region when rifampicin, or PCN was treated. Futher a trans-activation by SXR may demand other species-specific transcription factors.

• 한국환경보건학회지
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• 제30권3호
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• pp.230-238
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• 2004

The adverse health effects of a number of environment pollutions are related to the formation of free radicals. Induction of antioxidant defensive system in the response to an oxidative attack is an essential element of the cell to survive. CYP2E1 is easily induced by organic solvents and induces continuous formation of reactive oxygen species (ROS). ${\gamma}$-Glutamyltranspeptidase (${\gamma}$GT) plays an important role in glutathione metabolism and xenobiotic detoxification. To evaluate the characteristic of oxidative stress which induces GGT expression and to understand human antioxidant defensive response against oxidative stress induced by CYP2E1, we studied regulation of ${\gamma}$GT enzyme expression in response to various oxidative stresses in human HepG2 cells. The ${\gamma}$GT activity was not modified after exposure of acute oxidative stress inducing agents (ferric nitrilotriacetate, cumene hydroperoxide, ADP-Fe, O-tetradecanoylphorbol-13-acetate, tumor necrosis factor-alpha). To induce continuous exposure of cells to ROS, HepG2 cells were transfected by human CYP2E1 gene transiently. The CYP2E1 activity was verified with chlorzoxazone hydroxylation. Transfection of CYP2E1 showed continuous 60% increase in intracellular ROS and 240 % increase in microsomal ROS. CYP2E1 overexpressing cells showed increased ${\gamma}$GT activity (2.5-fold). The observed enhancement of ${\gamma}$GT activity correlated with a significant increase of ${\gamma}$GT mRNA (2.1-fold). Treatment with antioxidant strongly prevented the increase in ${\gamma}$GT activity. The CYP2E1 overexpression did not modify toxicity index and increased glutathione levels. These results show that continuous exposure of cells to ROS produced by CYP2E1 up-regulates ${\gamma}$GT; This may be one of the adaptive antioxidant responses of cells to oxidative insult. Present study also suggests that the induction of ${\gamma}$GT could be used as a marker of oxidative stress induced by exposure to organic solvents.

• Journal of Ginseng Research
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• 제29권4호
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• pp.159-166
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• 2005

Ginsenosides, a group of Panax ginseng saponins, exert the lowering effects of plasma cholesterol levels in animals. We had reported earlier that ginsenoside Rb2 upregulate low-density lipoprotein receptor (LDLR) expression via a mechanism that is dependent of the activation of sterol response element binding protein 2 (SREBP-2) expression. This study was conducted to determine the effects of ginsenoside Rb2 on the expression of the hepatic LDLR expression at cellular levels using HepG2 cells, and to evaluate whether the sterol response element binding protein 1 (SREBP-l) was involved in the regulation of LDLR expression. Incubation of HepG2 cells in serum-free medium supplemented with cholesterol $(10{\mu}g/ml)$ for 8 hours decreased the mRNAs of LDLR mRNA by $12\%$ and SREBP-l mRNA by $35\%$. Ginsenoside Rb2 antagonized the repressive effects of cholesterol and increased both LDLR and SREBP-l mRNA expression to 1.5- and 2-fold, respectively. Furthermore, Western blot and confocal microscopic analyses with SREBP-l polyclonal antibody revealed that ginsenoside Rb2 enhanced the maturation of the SREBP-1 from the inactive precursor form in ER membrane to the active transcription factor form in nucleus. These results suggest that ginsenoside Rb2 upregulates LDLR expression via a mechanism that is dependent of the activation of not only SREBP-2 expression, but also SREBP-1 expression and maturation, and also indicate that the pharmacological value of ginsenoside Rb2 may be distinguished from that of lovastatin which is reported that it upregulate LDLR through SREBP-2 only, not through SREBP-1.

• Advances in Traditional Medicine
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• 제3권3호
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• pp.141-146
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• 2003

The methanol extracts of the aerial parts, leaves and stem of Hypericum hookerianum were tested for in vitro cytotoxicity on selected normal and cancer cell lines and anti tumor activity using DLA cells. Cell viability and morphological changes were assessed. Among the three extracts tested, the stem extract of Hypericum hookerianum showed potent cytotoxicity against HEp-2 and RD cell lines. The $CTC_{50}$(concentration required to reduce viability by 50%) of this extract was found to be $2.02\;{\mu}g/ml$ for RD cell line, $10.25\;{\mu}g/ml$ for HEp-2 cell line and $100.06\;{\mu}g/ml$ for Vero cell line. In the clonogenic assay, no colony formation was observed up to a concentration of $100\;{\mu}g/ml$. In the short term cytotoxicity studies using DLA cells, 50% viability was observed in the concentration range of $50-100\;{\mu}g/ml$ for aerial parts, $100-200\;{\mu}g/ml$ for stem and more than $200\;{\mu}g/ml$ for leaf extracts of Hypericum hookerianum. In the long-term activity using HEp-2 cell line, no colony formation was observed over a concentration of 200 mg/ml for the stem extract. Hypericum hookerianum stem extract was fractionated into petroleum ether, chloroform, ethyl acetate and methanol soluble fractions. The petroleum ether and chloroform soluble fractions showed higher cytotoxic activity against HEp-2 cell line when compared to the other two fractions. The methanol stem extract of Hypericum hookerianum has the potential for further investigation in animal models to determine its anti-tumor activity and to identify its active principles.

• 한국자원식물학회지
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• 제25권5호
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• pp.647-651
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• 2012

참외 종자의 기능성을 규명하는 연구의 일환으로 항암활성을 조사하고자 하였다. 추출용매에 따른 항암활성을 조사하고자 헥산, 에탄올, 물 등 다양한 추출용매를 사용하여 참외 종자 추출물을 제조하였고, 가공에 따른 활성 증진효과를 알아보기 위해 볶음 처리하였다. 볶음 처리한 참외 종자는 볶음 처리하지 않은 종자보다 활성이 2~4배까지 증가하는 경향을 보여주었다. 추출용매에 따른 활성은 에탄올 추출물, 헥산 추출물, 물 추출물 순으로 항암활성이 나타났다. 인체 유래 5종(MCF-7, A549, AGS, HT-29, HepG2) 암 세포주를 이용하여 항암 활성의 조직 특이성을 알아보고자 하였다. 참외 볶음 종자 에탄올 추출물이 간암 세포주인 HepG2세포와 유방암 세포주인 MCF-7 세포에서 우수한 항암활성을 보여주었다. 용매분획법을 실시하여 제조한 분획물의 경우 에탄올 추출물보다 활성이 증가하였고, 비극성 분획물인 에칠아세테이트층이 극성 분획물인 부탄올층 보다 강한 항암활성을 보여주었다. 이상의 연구결과, 참외 볶음종자가 간암과 유방암 예방 및 치료 소재로서 개발될 가치가 있는 것으로 사료된다.

• 동의생리병리학회지
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• 제20권4호
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• pp.914-918
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• 2006

It was examined that the effect of fermented traditional wine made by using mycelium of Phellinus linteus on the expression of inflammation-related proteins in HepG2 cells. HepG2 cells were incubated with or without ertract of traditional wine (ETMP), then analyzed by microscopic observation, reverse transcription polymerase chain reaction (RT-PCR) and Western blot. The results of RT-PCR and Western blot analyses showed that the level of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and tumor necrosis factor $(TNF)-{\alpha}$ was induced by LPS, Dut the treatment of ETMP inhibited the expression of these proteins and its mRNAs. Besides, the results of Western blot analyses showed that the expression of nuclear $factor-{\kappa}Bp65$ and $inhibitory-{\kappa}B{\alpha}$ were also slightly affected by ETMP treatment. These results suggest that ETM P alleviate the expression of inflammation-related protein expressions and thus may be used as a functional alcoholic beverage.

• Journal of Ginseng Research
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• 제37권4호
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• pp.442-450
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• 2013

The aim of this study was to determine and compare the preventive effect of Korean White Ginseng and Red Ginseng on oxidative stress in $H_2O_2$-treated HepG2 cells. The roots of ginseng were extracted with 70% methanol and partitioned with butanol to obtain saponin fractions, which have been known as bioactive constituents of ginseng. 2',7'-Dichlorofluorescein diacetate (DCF-DA) assay and malondialdehyde (MDA) content were measured for evaluating intracellular reactive oxygen species (ROS) generation. Also, mRNA expressions and activities of antioxidant enzymes were analyzed to determine the antioxidant activity of saponin or non-saponin fractions of ginsengs. According to DCF-DA assay, $H_2O_2$-induced MDA release and ROS generation were significantly reduced by treatment with saponin fractions of white and red ginseng roots. Also, saponin fractions increased effectively intracellular antioxidant enzyme activities including catalase, glutathione peroxidase and superoxide dismutase in $H_2O_2$-treated HepG2 hepatoma cells. In general, red ginseng was more effective than white ginseng for reducing oxidative stress. These results indicate that administration of red ginseng may certainly contribute relatively stronger than white ginseng to prevent from damaging liver function by oxidative stress.

• Journal of Nutrition and Health
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• 제52권2호
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• pp.176-184
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• 2019

Purpose: Protein overloading in the endoplasmic reticulum (ER) leads to endoplasmic reticulum stress, which exacerbates various disease conditions. Emodin, an anthraquinone compound, is known to have several health benefits. The effect of emodin against palmitic acid (PA) - induced ER stress in HepG2 cells was investigated. Methods: HepG2 cells were treated with varying concentrations of palmitic acid to determine the working concentration that induced ER stress. ER stress associated genes such as ATF4, XBP1s, CHOP and GRP78 were checked using RT- PCR. In addition, the expression levels of unfolded protein response (UPR) associated proteins such as $IRE1{\alpha}$, $eIF2{\alpha}$ and CHOP were checked using immunoblotting to confirm the induction of ER stress. The effect of emodin on ER stress was analyzed by treating HepG2 cells with $750{\mu}M$ palmitic acid and varying concentrations of emodin, then analyzing the expression of UPR associated genes. Results: It was evident from the mRNA and protein expression results that palmitic acid significantly increased the expression of UPR associated genes and thereby induced ER stress. Subsequent treatment with emodin reduced the mRNA expression of ATF4, GRP78, and XBP1s. Furthermore, the protein levels of $p-IRE1{\alpha}$, $p-eIF2{\alpha}$ and CHOP were also reduced by the treatment of emodin. Analysis of sirtuin mRNA expression showed that emodin increased the levels of SIRT4 and SIRT7, indicating a possible role in decreasing the expression of UPR-related genes. Conclusion: Altogether, the results suggest that emodin could exert a protective effect against fatty acid-induced ER stress and could be an agent for the management of various ER stress related diseases.

• 대한약침학회지
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• 제22권1호
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• pp.41-48
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• 2019

Objective: The aim of the study was to evaluate the hepatoprotective activity of the bark extract (Ethanol: Water) in the ratio of (3:1) of Rhizophora mucronata (BERM) by intoxicating the $HepG_2$ cell lines with different toxicants viz, $CCL_4$, Ethanol and Paracetamol with different concentrations of the extract were used. The $HepG_2$ cell lines were subjected to MTT Assay for studying the cytotoxicity. Methods: $HepG_2$ cells were plated using 96 well plate in 10% bovine serum, exposed to different toxicants viz, 2% $CCl_4$, 60% Ethanol and 14 mM Paracetamol respectively. The various test concentrations (18.85, 37.5, 75, 150 and $300{\mu}g/ml$) of bark extract of Rhizophora mucronata was added and incubated for 24 hours. Medium was removed after incubation period and 0.5 mg/ml MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) was added and again incubated for 4 hours at 37oC. Then MTT was removed the crystals was dissolved in DMSO and absorbance was measured at 570 nm. Results: The result showed that dose dependent increase in percentage of viability at the doses of 18.85, 37.5, 75, 150, $300{\mu}g/ml$. Te results for the $CCl_4$ intoxicated, at $300{\mu}g/ml$ of the concentration of the extract, the % of viable cells was found out to be 99.6%, for Ethanol intoxicated, 97.67%, and Paracetamol induced, 75.37%, IC50 was $21.53{\mu}g/ml$, $12.61{\mu}g/ml$ and $21.42{\mu}g/ml$ respectively. Conclusion: Thus, we conclude that, the extract possesses defensive effect against different toxicants and can be used as an alternate drug for hepatotoxicity.