• Advances in Traditional Medicine
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• v.6 no.1
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• pp.45-52
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• 2006

The methanol extracts of the roots, root bark and stem of Berberis tinctoria, were investigated for their hepatoprotective activity against carbon tetrachloride $(CCl_4)$ induced toxicity in freshly isolated rat hepatocytes, HEp-G2 cells and animal models. The methanol extracts were able to significantly normalise the levels of aspartate amino transferase, alanine aminotransferase, alkaline phosphatase, triglycerides, total proteins, albumin, total bilirubin and direct bilirubin, which were altered due to $CCl_4$ intoxication in freshly isolated rat hepatocytes and also in animal models. The anti-hepatotoxic effect of the methanol extracts in vitro were observed at $600\;-\;1,000\;{\mu}g/ml$ concentrations. A dose dependent increase in the percentage viability was observed when $CCl_4$ exposed HEp-G2 cells were treated with different concentrations of the methanol extracts. The highest percentage viability of HEp-G2 was observed at a concentration of $1,000\;{\mu}g/ml$. The results from the present investigations also indicate good correlation between the in vivo and in vitro studies.

• Parasites, Hosts and Diseases
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• v.61 no.4
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• pp.388-396
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• 2023

Entamoeba histolytica is an enteric tissue-invasive protozoan parasite causing amoebic colitis and liver abscesses in humans. Amoebic contact with host cells activates intracellular signaling pathways that lead to host cell death via generation of caspase-3, calpain, Ca²⁺ elevation, and reactive oxygen species (ROS). We previously reported that various NADPH oxidases (NOXs) are responsible for ROS-dependent death of various host cells induced by amoeba. In the present study, we investigated the specific NOX isoform involved in ROS-dependent death of hepatocytes induced by amoebas. Co-incubation of hepatoma HepG2 cells with live amoebic trophozoites resulted in remarkably increased DNA fragmentation compared to cells incubated with medium alone. HepG2 cells that adhered to amoebic trophozoites showed strong dichlorodihydrofluorescein diacetate (DCF-DA) fluorescence, suggesting intracellular ROS accumulation within host cells stimulated by amoebic trophozoites. Pretreatment of HepG2 cells with the general NOX inhibitor DPI or NOX2-specific inhibitor GSK 2795039 reduced Entamoeba-induced ROS generation. Similarly, Entamoeba-induced LDH release from HepG2 cells was effectively inhibited by pretreatment with DPI or GSK 2795039. In NOX2-silenced HepG2 cells, Entamoeba-induced LDH release was also significantly inhibited compared with controls. Taken together, the results support an important role of NOX2-derived ROS in hepatocyte death induced by E. histolytica.

• 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.

• KSBB Journal
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• v.23 no.5
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• pp.381-386
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• 2008

Lipophilic ammonia is toxic gas and can easily diffuse across cell membranes. Excess ammonia is implicated in the pathogenesis of several metabolic disorders including hepatic encephalopathy and may result in the death. The purpose of this study was to clarify the inhibition effect of metronidazole on liver cell damage due to ammonia in human Hep G2 cell and rat hepatocytes. The effects of metronidazole were studied in ammonium chloride treated human Hep G2 cell (75 mM) and rat hepatocyte (100 mM) following $0.1{\mu}M$ metronidazole treatment. In MTZ+AC group, cell viabilities increased prominently and LDH activities decreased over 25% than AC group. Furthermore, ammonia level according to ammonium chloride treatment reduced over 30% and lipid peroxidation as an index of cell membrane damage decreased more than twice. By comparison with control, catalase activity showed more than 30% reduction in AC group while less than 10% reduction in MTZ+AC group, respectively. In addition, MTZ+AC group showed the similar cell structure as control in cell morphology study by using light microscope, and represented fluorescent intensity decrement compared with AC group in fluorescent microscopic study with avidin-TRITC fluorescent dye. And cleaved PARP expression due to ammonia reduced twofold or more in MTZ+AC group. As the results suggest, metronidazole may protect the liver cell by inhibiting cell damages due to ammonia and be used for an effective antagonist of ammonia in hyperammonemia.

• BMB Reports
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• v.30 no.4
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• pp.269-273
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• 1997

Heptocvte-specific expression induced by Hepatitis B virus (HBV) enhancer 2-core gene promoter was examined in various hepatocyte and non-hepatocyte cell lines. using non-viral and retroviral vector systems in which chloramphenicol acetyltransferase (CAT) is used as a reporter. The non-viral plasmid containing the HBV enhancer 2-core promoter exhibited 22 and 66% of CAT activities in hepatoma cell lines. HepG2 and Hep3B, respectively when compared with CAT activity expressed by CMV promoter. The CAT activities, however. were found to be marginal in other tested hepatoma cell lines as well as mouse primary hepatocytes and non-hepatocytes. The HBV enhancer 2 located upstream the CMV promoter did not affect the CMV promoter activity nor provided hepatocyte-specific expression. Transfection of retroviral plasmid DNA containing the HBV enhancer 2-core promoter as an internal promoter exhibited high and specific CAT expression in HepG2 and Hep3B cell lines but the activity value was 5 to 10 fold lower than the non-viral plasmid with identical promoter. These results suggest that the usage of HBV enhancer 2-core promoter for liver specific expression is limited to certain vectors and hepatocyte cell lines.

• The Korea Journal of Herbology
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• v.23 no.3
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• pp.149-154
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• 2008

Objectives : The purpose of this study is to investigate the biological Effect of multi-herbal drug 'KOCO-P1' on human hepatocyte HepG2 cells. Methods : Multi-herbal drug 'KOCO-P1' was composed of Ginseng Radix, Astragali Radix, Polygonati Rhizoma, Liriopis Tuber, and Scrophulariae Radix. Cytotoxicity and cytoprotective activity of KOCO-P1 was verificated by MTT assay. And antioxidative effect of KOCO-P1 against EtOH, Nicotine was inspected by Hydroperoxide assay. Results : KOCO-P1 showed no cytotoxicity on HepG2 cells for 24, 48 hours. KOCO-P1 at 50 ${\mu}g/mL$ reduced the production of H2O2 in HepG2 cells by EtOH. KOCO-P1 at 50 ${\mu}g/mL$ reduced the production of $H_2O_2$ in $HepG_2$ cells by Nicotine. Conclusions : KOCO-P1 at the low concentration could be supposed to have antioxidative effect on human hepatocyte with no cytotoxicity.

• Korean Journal of Environmental Agriculture
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• v.28 no.4
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• pp.462-467
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• 2009

Exposure of formaldehyde (FA), one of the major compounds in pesticides and in the onset of sick house syndrome, has been implicated in the development of diverse diseases. Liver is a very important organ to body metabolism and drug detoxification. Apotosis of hepatocytes is associated with the onset of liver diseases such as hepatitis. However, the apoptotic effect of FA in hepatocytes is not clear. Therefore, this study was conducted to investigate the effect of FA on the apoptosis in HepG2 cells, a human hepatocyte cell line. As a result, FA (> $500\;{\mu}M$) decreased cell viability and increased lactate dehydrogenase activity in HepG2 cells, which was blocked by the treatment of vitamin E and N-acetylcysteine (NAC). In addition, FA decreased glutathione (GSH) contents and Bcl-2 levels, while increasing lipid peroxide formation and Bax levels. It also cleaved caspase-3 form, which was blocked by the treatment of vitamin E and NAC. It is insisted that FA induced apoptosis via oxidative stress in human hepatocytes.

• Applied Biological Chemistry
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• v.40 no.6
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• pp.577-582
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• 1997

Effects of four citrus flavonoids, hesperidin, naringin and their aglycones on phosphatidate phosphohydrolase(PAP, EC 3.1.3.3) activity were examined using isolated rat microsomes as an enzyme source. In addition, these flavonoids were tested to see whether they exert any influence on the proliferation and growth in cultured human hepatocytes HepG2 cells. Flavonoids at concentration up to $10{-4}M$ had no significant effect on the number of cells and cell proliferation by MTT cell growth assay method, whereas aglycone flavonoids, hesperetin and narigenin, at concentration of $10{-3}M$ significantly inhibited cell proliferation. Hesperetin inhibited PAP activity in a dose-dependent manner starting at concentration of $10{-5}M$. Narigenin at concentration of $10{-2}M$ inhibited PAP activity markedly, while the other flavonoids did not show any significant effect. The present study, therefore, demonstrated that aglycone flavonoids exerted portent effects on PAP activity and on cell proliferation.

• Journal of the East Asian Society of Dietary Life
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• v.25 no.1
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• pp.99-110
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• 2015

The purpose of this study was to evaluate the inhibitory effects of commonly consumed vegetables in Korea on lipid accumulation and production of pro-inflammatory cytokines related to obesity/metabolic syndrome. Using KNHANES (Korea National Health and Nutrition Examination Survey) raw data ($1^{st}$; 1998, $5^{th}$; 2010, 2011) and a literature search, we selected vegetables for study. Edible portions of samples were prepared, ethanol-extracted, and then freeze-dried. 3T3-L1 adipocytes and HepG2 hepatocytes cells were used as in vitro models. Lipid accumulation determined by Oil-red O staining showed that all samples except bracken had inhibitory effects on lipid accumulation in 3T3-L1 adipocytes. Especially, crown daisy and mugwort effectively reduced accumulation of lipids, and their inhibition rates were more than 60% of the control group. Young pumpkin, leeks, crown daisy, and mugwort showed significantly decreased MCP-1 levels compared to the control group. However, adiponectin protein level did not increase in the vegetables experimental group. In HepG2 hepatocytes, all samples showed inhibitory effects on lipid accumulation at one of the two concentrations. Although adiponectin protein levels did not increase, MCP-1 protein levels decreased in adipocytes. Further, lipid accumulation in adipocytes and hepatocytes decreased. In conclusion, all samples showed one or more improved obesity/metabolic syndrome indicators. Among them, young pumpkin, leeks, crown daisy, and mugwort were selected as the most effective portions of vegetables based on improvement of obesity/metabolic syndrome-related indicators.

• Archives of Pharmacal Research
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• v.27 no.9
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• pp.944-946
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• 2004

Phytochemical investigation of the aqueous extract of ~he roots of Agrimania pilosa Ledeb. (Rosaceae), as guided by hepatoprotective activity in vitro, furnished two isocoumarins, agri-monolide (1) and agrimonolide 6-O-$\beta$-D-glucoside (3), and (＋)-catechin (2). Compound 1 showed hepatoprotective effects on both tacrine-induced cytotoxicity in human liver-derived Hep G2 cells and tert-butyl hydroperoxide-induced cytotoxicity in rat primary hepatocytes with EC$_{50}$ values of 88.2$\pm$2.8 and 37.7$\pm$1.6 $\mu$M, respectively.y.