• The Journal of Internal Korean Medicine
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• v.25 no.1
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• pp.28-45
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• 2004

Objectives : The main purpose of this study is to evaluate the effect of Injinchunggan-tang on $TNF-{\alpha}$ signal transmission system. Materials and Methods : We analyzed the following with quantitative RT-PCR method; the effect of Injinchunggan-tang on secretion of $TNF-\alpha$ mRNA/protein and stability, the effect on gene revelation that consists of signal transmission system (TRAIL, NIK, A20, TRADD, RAIDD, RIP TNFR-I, TNFR-II, TRAF1, TRAF2, FADD), the one on activation of p38, Erk1/2 MAPK and the rate of nuclear $NF-{\kappa}B/cytosolic\;NF-{\kappa}B$ in HepG2 cell. We also analyzed the inhibitory effect of Injinchunggan-tang on the apoptosis of HepG2 cell that $TNF-{\alpha}$ induces and the $NF-{\kappa}B$ restraint effected by transfection of $I{\kappa}B{\Delta}N$ through tryphan blue exclusion assay. Results : Injinchunggan-tang prohibits revelation of $TNF-{\alpha}$ mRNA in HepG2 cell and the creation of protein. However, it has no effect on the stability of $TNF-{\alpha}$ mRNA. While it did not have any effect on the generation of TRAIL, NIK, A20, TRADD, RAIDD and RIP genes, Injinchunggan-tang reduces the revelation of TNFR-I, TNFR-II, TRAF1, TRAF2 and FADD genes. It has been confirmed that Injinchunggan-tang restraints the revelation of $TNF-{\alpha}$ mRNA that is promoted by ethanol, acetaldehyde, lipopolysaccharide, in proportion to the treatment density and time. It activated $NF-{\kappa}B$ of HepG2 cell and promoted activation of $NF-{\kappa}B$ that is occurred by $TNF-{\alpha}$. It has been observed that the restraint effect against the $TNF-{\alpha}$ inducing apoptosis is lost when it is intercepted the function of $NF-{\kappa}B$ in HepG2 cell. Conclusion: It has been confirmed that Injinchunggan-tang has restraining effect against the revelation of $TNF-{\alpha}$ and mRNA that is constituent element of TNF-a signal transmission system. It also has been revealed that it restraints the activation of p38, Erk1/2 by $TNF-{\alpha}$. Through this prohibiting effect, it is inferred that it restraints signal transmission among various cells that are related to inflammation reaction. Meanwhile, Injinchunggan-tang protects liver cell from apoptosis that is caused by $TNF-{\alpha}$, by maintaining the activating function for $NF-{\kappa}B$.

• Journal of Life Science
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• v.29 no.10
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• pp.1144-1151
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• 2019

Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease associated with various metabolic syndromes, such as obesity, dyslipidemia, hypertension, and diabetes. Cudrania tricuspidata is a medicinal plant distributed widely in Asia and has been used in clinical practice to treat various diseases. The aim of this study is to determine the lipid-lowering effects of C. tricuspidata fruit extract (CTE) using a cell model induced by free fatty acids (FFAs). HepG2 cells were exposed to 1mM FFAs (palmitic acid:oleic acid = 2:1) for 24 hr to simulate the conditions of NAFLD in vitro. CTE attenuated the increases of lipid accumulation, intracellular triglyceride, and cholesterol content and inhibited 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) activity in the HepG2 cells in a dose-dependent manner. Also, CTE inhibited the protein expression of lipogenesis-related genes, such as sterol regulatory element-binding protein-1/-2 (SREBP-1/-2), fatty acid synthase (FAS), and stearoyl CoA desaturase-1 (SCD-1) in FFAs-induced lipid accumulation in HepG2 cells. In addition, CTE-induced adenosine monophosphate-activated protein kinase (AMPK) phosphorylation in HepG2 cells. These results suggest that CTE attenuates hepatic lipid accumulation by inhibiting lipogenesis through the modulation of the AMPK signaling pathway on FFAs-induced lipogenesis in HepG2 cells and may potentially prevent NAFLD.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.2
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• pp.427-430
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• 2004

This study was to investigate the cell cycle arrest effect and its mechanism of Radix Aconiti (RA) extract in HepG2 human hepatoma cells. We used the Flow Cytometer to investigate the effects on cell cycle arrest in RA extract-treated HepG2 cells. And protein levels involved in cell cycle progression such as p53, p21, and p21 are detected by Western blotting method. RA extract induced cell cycle arrest as confirmed by increase of G0/G1 cell population, and the mechanisms were related with up-regulation of p53, p21, p27 protein expressin in HepG2 cells. These results suggest that RA may be a valuable agent for the therapeutic intervention of human hepatomas.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.6
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• pp.2619-2623
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• 2014

Aim: To investigate signaling pathways for reversal of EGF-mediated multi-drug resistance (MDR) in hepatocellular carcinoma (HCC) models. Materials and Methods: HCC MDR cell strain HepG2/adriamycin (ADM) and SMMC7721/ADM models were established using a method of exposure to medium with ADM between low and high concentration with gradually increasing concentration. Drug sensitivity and reversal of multi-drug resistance by EGF were determined and the cell cycle distribution and apoptosis were analyzed by flow cytometry. Phosphorylation of ERK1, ERK2, ERK5 and expression of Bim were detected by Western blotting. Results: The results showed that HepG2/ADM and SMMC7721/ADM cells were resistant not only to ADM, but also to multiple anticancer drugs. When used alone, EGF had no anti-tumor activity in HepG2/ADM and SMMC7721/ADM cells in vitro, while it increased the cytotoxicity of ADM. EGF induced cell apoptosis and G0/G1 phase cell cycle arrest in HepG2/ADM And SMMC7721/ADM cells, while enhancing activity of p-ERKs and up-regulated expression of BimEL. Conclusions: EGF might enhance the chemosensitivity of HepG2/ADM and SMMC7721/ADM cells via up-regulating p-ERKs and BimEL protein.

• BMB Reports
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• v.29 no.1
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• pp.73-80
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• 1996

The effect of albumin on phosphatidylcholine (PC) metabolism in Hep-G2, 3T3-H.ras, and 3T3 cells pre-labelled with [Me-$^3H$]choline was studied. The [$^3H$]choline was more efficiently taken up and incorporated into cellular phospholipids in 3T3-H.ras cells than in Hep-G2 and 3T3 cells. In each of the three cell lines, most of the [$^3H$]choline metabolized into the phospholipids was incorporated into PC and only minor was incorporated into lysophosphatidylcholine (LPC). Bovine serum albumin stimulated the release of [$^3H$]LPC and [$^3H$]PC from each of the three cell lines pre-labelled with [$^3H$]choline. [$^3H$]PC was also released in the absence of albumin but [$^3H$]LPC was not. The efficiency of LPC secretion represented as the proportion of medium [$^3H$]LPC to cellular [$^3H$]choline lipid during a chase period is approximately 9 to 14 times greater in 3T3 cells compared with the transformed 3T3-H.ras and Hep-G2 cells. A similar comparison of published data for rat hepatocytes with Hep-G2 shows secretion to be 35~75 times greater from the rat hepatocytes than from Hep-G2. Also, PC secretion from 3T3 cells was 1.6 times more effective than from 3T3-H.ras, whereas rat hepatocytes secrete PC 2.8~3.8 times more effectively than does Hep-G2. The measurement of specific radioactivity of cellular PC in pre-labelled 3T3 cells showed it to be similar to that of the secreted PC. However, the specific radioactivity of secreted LPC was markedly lower than that of the cellular PC, which suggests that LPC is being secreted from a PC pool distinct from that used for PC secretion.

• The Journal of Internal Korean Medicine
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• v.38 no.3
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• pp.367-375
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• 2017

Objective: To investigate the effect of Artemisiae Iwayomogii Herba, Curcumae Radix, and Aurantii Fructus Immaturus complex extract (ACA) on dyslipidemia-related factor expression and anti-oxidation in HepG2 cells. Method: After treatment with ACA in the HepG2 cells, DPPH, ABTS radical scavenging activity, ROS production, and glutathione (GSH) production were measured. The free fatty acid, lipid peroxidation (MDA), ACAT1, and HMG-CoA reductase mRNA expression were measured in the HepG2 cells after treatment with ACA. Results: 1. DPPH, ABTS radical scavenging activity increased in an ACA concentration-dependent manner. 2. ACA significantly decreased ROS production in comparison to the control group. 3. ACA significantly increased glutathione production. 4. ACA significantly decreased free fatty acid and lipid peroxidation (MDA) in the HepG2 cells. 5. ACA decreased the mRNA expression of ACAT1 and HMG-CoA reductase. Conclusion: These results suggest that Artemisiae Iwayomogii Herba, Curcumae Radix, and Aurantii Fructus Immaturus complex extract (ACA) inhibits dyslipidemia-related factor expression and that it is effective in anti-oxidation. A future in vivo experiment with ACA is needed to investigate the effect on anti-dyslipidemia. It is expected that ACA is effective in anti-dyslipidemia and applied to cardiovascular disease, ischemic heart disease, stroke, etc.

• Herbal Formula Science
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• v.13 no.2
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• pp.111-123
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• 2005

The purpose of this study was to investigate the anticancer effects of Caesalpiniae Lignum (Somok) on HepG2 cells, a human liver cancer cell line. To study the cytotoxic effect of Caesalpiniae Lignum methanol extract (CL-MeOH) on HepG2 cells, the cells were treated with various concentrations of CL-MeOH and then cell viability was determined by XTT reduction method and trypan blue exclusion assay. CL-MeOH reduced proliferation of HepG2 cells in a dose-dependent manner. To confirm the induction of apoptosis, HepG2 cells were treated with various concentrations of CL-MeOH. The activation of caspase 3 and the cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, was examined by western blot analysis. CL-MeOH decreased procaspase 3 level in a dose-dependent manner and induced the clevage of PARP at concentration> $200{\mu}/ml$. Mitogen-activated protein (MAP) kinase signaling cascades are multi-functional signaling networks that influence cell growth, differentiation, apoptosis, and cellular responses to stress. CL-MeOH-induced MAPK activation was examined by Western blot for phosphorylated ERK, p38 and JNK. CL-MeOH significantly increased p38 phosphorylation and JNK phosphorylation in a dose-dependent manner. Inhibition of p38 function using the selective inhibitor SB20358O results in inhibition of apoptosis by CL-MeOH. These results suggest that CL-MeOH-induced apoptosis is MAP kinase-dependent apoptoric pathway. These results suggest that CL-MeOH is potentially useful as a chemotherapeutic agent in human liver cancer.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.45 no.6
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• pp.553-560
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• 2012

Hizikia fusiformis, a brown alga that is widely consumed in Korea, Japan, and China, possesses a number of potentially beneficial compounds, including antioxidants and anticoagulants. However, the molecular mechanisms of H. fusiformis in hepatoma cells have not been elucidated. This study investigated the antiproliferative effect and mechanism of action of a glycoprotein from H. fusiformis (HFGP) in HepG2 human hepatoma cells. In an MTS assay, 25 ${\mu}g/mL$ HFGP inhibited the proliferation of HepG2 cells by $52.36{\pm}2.37%$. HFGP caused the dose-dependent growth inhibition of HepG2 cells by inducing apoptosis and a sub-G1 phase arrest. The antiproliferative activity of HFGP was confirmed based on the expression of several apoptosis-related proteins, which was assessed by Western blot analysis. The expressions of Fas, Fas-associated death domain protein, Bax, and Bad was significantly up-regulated in HFGP-treated cells, and HFGP induced the translocation of Bax to mitochondria and the release of cytochrome c into the cytosol. Therefore, HFGP might be useful in the treatment of liver cancer.

• Korean Journal of Food Science and Technology
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• v.54 no.4
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• pp.398-402
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• 2022

Black chokeberry (Aronia melanocarpa), a rich source of polyphenols, exerts hypocholesterolemic effects. However, little is known about its effects on the regulation of the hepatic cholesterol metabolism and the underlying mechanisms. In the present study, the effects of polyphenol-rich black chokeberry extract (CBE) on hepatic cholesterol metabolism were investigated by measuring the expression of genes involved in the absorption, de novo synthesis, and efflux of cholesterol in HepG2 cells. There was a significant reduction in the expression levels of genes involved in cholesterol metabolism, the low-density lipoprotein receptor, 3-hydroxy-3-methylglutaryl coenzyme A reductase, and sterol regulatory element-binding protein 2, in CBE-treated HepG2 cells. Meanwhile, CBE increased the expression levels of genes involved in cholesterol and bile acid efflux. The expression levels of mitochondrial fatty acid oxidation genes increased, whereas those of lipogenic genes decreased following CBE treatment. These data suggest that the consumption of black chokeberry may be beneficial for the prevention of hypercholesterolemia.

• The Journal of Internal Korean Medicine
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• v.26 no.2
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• pp.320-332
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• 2005

Objectives: The aim of the experiment is to identify any anti-tumor effects of Duchesnea indica(Andr.) Focke on stomach, liver, urinary bladder, prostate and kidney cancer cells. Materials & Methods: For cancer cells, AGS stomach, Hep3B and Hep3G2 liver, HT-1197, HT-1376 urinary bladder, PC3 prostate, and A-704 kidney cancer cells, all obtained from Korean Ce 11 Line Bank, were used. The boiled extract of Duchesnea indica(Andr.) Focke (10 and 20 microliters) was injected into cultures, and the cultures were observed at 0, 6 and 12 hours, and from then on at 12 hours intervals up to 72 hours. The destruction of stomach, liver, urinary bladder, prostate and kidney cancer cells were measured through Trypan blue exclusion testing. The suppresion on viability of stomach, liver, urinary bladder, prostate and kidney cancer cells was measured via MTT assay. Anti-cancer mechanisms were assessed by analyzing the cell cycle. Results: In morphologic change, AGS, Hep3B, HepG2 showed the withdrawn and floating appearance that is typical in cellular impairment. The destruction of AGS, HT-1197, HT-1376, A-704, PC-3, Hep3B and HepG2 cancer cells in each test group was greater than that in the control group to a statistically significant degree. The suppression on viability of AGS, HT-1197 and Hep3G in each test group was greater than that in the control group to a statistically significant degree. Analysis of the cell cycle after injection of D... Focke showed inhibition of cell division in all test groups(AGS, Hep3B, HepG2, HT-1197, HT-1376, PC3, A-704). Conclusions: The results of this experiment suggest that Duchesnea indica(Andr.) Focke has statistically significant anti-tumor effects on stomach, urinary bladder, kidney, prostate and liver cancer, of which stomach and liver cancer are prominently significant. This in vitro experiment supports a role for Duchesnea indica(Andr.) Focke as a potential cancer treatment, but progressive research on Duchesnea indica(Andr.) Focke and its anti-tumor effects is needed to develop a practical application for it in cancer treatment.