• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.363-364
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• 2003

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2001.05a
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• pp.185-186
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• 2001

Recently, industrial activities have increased atmospheric concentration of sulfur and nitrogen oxides, resulting in acidification in the environments. In addition, acidification accelerates the mobilization of metals that are toxic to fish and increases their concentrations in the aquatic environment. Increased metals may interfere with reproductive physiology in fish. Al and Cd are such metals that impaired the preduction of Vitellogenin (VTG), a egg yolk precursor proteins. (omitted)

• Journal of Yeungnam Medical Science
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• v.6 no.2
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• pp.133-140
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• 1989

To investigate recovery, growth, and activity of hepatocyte in primary culture after cell separation, the authors followed up the marker enzyme activities of golgi complex, mitochondria and biologic membrane. Thiamine pyrophosphatase, the marker enzyme of golgi complex, activity approached the level of long term culture at 4th day. Succinate dehydrogenase, the marker enzyme of mitochondria, activity decreased with time, then it maintained constant level after 4th day. Alkaline phosphatase, the marker enzyme of biological membrane, activity increased from 3rd day, and after 5th day it showed strong reaction. These data suggested that hepatocytes were stabilized and recovered normal activity 4 day after cell separation, but the main secretory function was speculated to be reduced in culture.

• Korean Journal of Environmental Agriculture
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• v.22 no.2
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• pp.94-99
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• 2003

The objective of present study was to investigate the antioxidative and hepatoprotective effects of selenium on cadmium-induced toxicity and lipid peroxidation in rat hepatocyte primary culture. To do this, two separate experiments were conducted. In Experiment 1, primary cultures of rat hepatocytes were incubated for 6 hr in the presence of various concentrations (1, 10, 50, 100, and $500\;{\mu}M$) of cadmium chloride. Cytotoxicity and lipid peroxidation were evaluated using the MTT assay and TBARS assay, respectively. Antioxidative and hepatoprotective effects were determined by measuring the activity of GOT and GSH-Px, respectively. Cell viability was reduced and lipid peroxidation was increased by cadmium in dose-dependent manners. There was significantly negative correlation (r=-0.943, p<0.01) between cell viability and lipid peroxidation GOT activity was increased and GSH-Px activity was decreased by cadmium at the concentration of $50\;{\mu}M$. In Experiment 2, primary cultures of rat hepatocytes were incubated for 6hr in the presence of 100\;{\mu}M$ of cadmium chloride and various concentrations (0.01, 0.1 and 1 ppm) of sodium selenite to assess the effect of selenium on cadmium-induced toxicity and lipid peroxidation. Cell viability and GSH-Px activity were increased by sodium selenite at the concentration of 1 ppm Whereas, lipid peroxidation and GOT activity were reduced by 0.1 ppm of sodium selenite. These results demonstrate that selenium has an antioxidative and hepatoprotective potentials against cadmium.

• Journal of fish pathology
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• v.23 no.2
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• pp.245-254
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• 2010

Effects of heavy metal ions on the gene expression of cytochrome P4501A (CYP1A) were examined in cultured eel hepatocytes. When the expression of CYP1A mRNA was measured by RT-PCR after incubation of eel hepatocytes with benzo[$\alpha$]pyrene (B[$\alpha$]P) at concentrations of 10-8~10-5 M, the CYP1A expression increased with B[$\alpha$]P treatment in a dose dependent manner, showing significant increase at concentrations more than 10-7 M. When the eel hepatocyte was treated with cadmium (10-6 and 10-5 M), the expression of CYP1A was inhibited and especially at higher concentration (10-5 M). The inhibition of CYP1A expression by cadmium was also observed in cells treated with B[$\alpha$]P. In another study, effects of heavy metal ions on the expression of CYP1A were examined in cultured hepatocytes isolated from eel which was treated previously with B[$\alpha$]P in vivo. Hepatocytes isolated from the liver of eel taken at 48 hours after injection of B[$\alpha$]P (10 mg/kg) were cultured for 2 days with cadmium, copper, lead or zinc (10-6 and 10-5 M). The expression of CYP1A was found to be suppressed by the metal ions compared with the control in which CYP1A was induced with previous treatment of B[$\alpha$]P in vivo. The present results may provide an important basic information for studying the effects of heavy metal ions on CYP1A expression in other species of fish and studying toxicological mechanisms of heavy metal ions in aquatic livings.

• BMB Reports
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• v.47 no.8
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• pp.457-462
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• 2014

The pathogenesis of non-alcoholic steatohepatitis (NASH) is not fully understood. In the present study, both in vitro and in vivo vimentin expression and secretion in NASH were investigated. The exposure of palmitate and lipopolysaccharide (LPS) to HepG2 cells enhanced caspase-3 activity and vimentin expression, respectively. The combined effects of both treatments on vimentin expression and caspase-3 activation appeared to be synergic. In contrast, blockade of caspase-3 activity by zVADfmk resulted in a significant reduction of cleaved vimentin and secreted vimentin into the culture supernatant. Similarly, lipid accumulation and inflammation occurred in mice fed a methionine-choline-deficient diet; thus, vimentin expression and serum cleaved vimentin levels were increased. However, vimentin was not significantly upregulated, and no cleavage occurred in mice fed a high-fat diet. It was conclusively determined that lipid accumulation in hepatocytes induces apoptosis through a caspase-3 dependent pathway; whereas, LPS stimulates vimentin expression, leading to its cleavage and secretion. Increased vimentin fragment levels indicated the existence of substantial hepatocellular death via an apoptotic mechanism.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.129-130
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• 1994

The science of toxicology is the understanding of the mechanisms by which exogenous agents produce deleterious effects in biological systems. The actions of chemicals such as drugs are ultimately exerted at the cellular and gene levels. Over the past decade. several in vitro alternative methods such as cultured cells for assessing the toxicity of various xenobiotics have been proposed to reduce the use of animals. In this workshop three advanced methods will be presented. These methods are novel important models for toxicologic studies. Dr. Tabuchis group has establishcd two immortalized gastric surface mucosa cell lines from the pminary cultore of gastric fundic mucosal cells of adult transgenic mice harboring a temperature sensitive simian virus 40 large T-anugen gene. As the immortalized cell lines of various tissues possess unique characteristics to maintain their normal functions for several months, these cell lines are extremely useful for not only toxicity testing but also pharmacological screening in new drug development. Professor Funatsu have studied the formation of spherical multicelluar aggregates of adult rat hepatocytes(spheroid) having tissue like structure. The sphcroid shown thre is a prototype module of an artificial liver support system. Thus, the urea synthesis activity of the artificial liver was maintained at least to days in 100％ rat blood plasma. Dr. Takezawa and his coworkers have developed a novel culture system of multicellular spheroids considered 〃organoids〃 by utilizing a thermo-responsive polymer as a substratum of anchorage dependent cells. His final goal is to reconstitute the organoids of various normal organs, e.g., liver, skin etc. and also abnormal deseased organs such as tumor.

• BMB Reports
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• v.40 no.6
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• pp.1002-1008
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• 2007

PreS domain of Hepatitis B virus (HBV) surface antigen is a good candidate for an effective vaccine as it activates both B and T cells besides binding to hepatocytes. This report deals with overexpression and purification of adr subtype of surface antigen that is more prevalent in Pakistan. PreS region, comprising 119 aa preS1 region plus a 55 aa preS2 region plus 11 aa from the N-terminal S region, was inserted in pET21a+ vector, cloned in E. coli $DH5\alpha$ cells and expressed in E. coli BL21 codon+ cells. The conditions for over expression were optimized using different concentrations of IPTG (0.01-5 mM), and incubating the cells at different temperatures (23-$41^{\circ}C$) for different durations (0-6 h). The cells were grown under the given optimized conditions (0.5 mM IPTG concentration at $37^{\circ}C$ for 4 h), lysed by sonication and the protein was purified by ion exchange chromatography. On the average, 24.5 mg of recombinant protein was purified per liter of culture. The purified protein was later lyophilized and stored at $-80^{\circ}C$.

• Journal of the East Asian Society of Dietary Life
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• v.5 no.1
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• pp.21-27
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• 1995

The protective effect of omija methanol extract on benzo(a)pyrene induce liver injury was studied in rats in vitro and in vivo. In vitro experiment, primary cultured hepatocytes(5${\times}$105cells/$m\ell$) were cultured for 20∼24 hours after adding omija methanol extract(5.1$\mu\textrm{g}$/$m\ell$) and B(a)P(50$\mu\textrm{m}$) in culture medium. In vivo experiment, omija methanol extract(0.1g/kg/day, per os) was administered for 7days and B(a)P(0.1mg/kg body weight, intraperitoneally) was given to the rats after the last administration of extract. Omija methanol extract significantly recovered serum enzyme activities(AST, ALT and LDH) and lipid contents(total cholesterol, triglyceride and HDL-cholesterol) changed by benzo(a)pyrene (B(a)P) to normal levels in vivo. In vitro experiment, as a result of 3-(4, 5-dimethlythiazol-2-yl)-2, 5-diphenyl tetrazolium bromide(MTT) assay, omija methanol extract showed a little hepatotoxicity compared with group I (normal) but significantly recovered enzyme activities(AST, ALT and LDH) changed by B(a)P in comparison to group IIadministered B(a)P only. It was suggested that omija methanol extract has a protective effect on liver injury induced by B(a)P.

• Biomolecules & Therapeutics
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• v.29 no.2
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• pp.205-210
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• 2021

Over 30 million prescriptions of NSAIDs (non-steroidal anti-inflammatory drugs) are issued every year. Considering that these drugs are available without a prescription as over the counter (OTC) drugs, their use will be astronomical. With the increasing use of NSAIDs, their adverse effects are drawing attention. Especially, stomach bleeding, kidney toxicity, liver toxicity, and neurological toxicity are reported as common. Ibuprofen, one of the extensively used NSAIDs along with aspirin, can also induce liver toxicity, but few studies are addressing this point. Here we examined the liver toxicity of ibuprofen and investigated whether co-exposure to ethanol can manifest synergistic effects. We employed 2D and 3D cultured human hepatoma cells, HepG2 to examine the synergistic hepatotoxicity of ibuprofen and alcohol concerning cell viability, morphology, and histology of 3D spheroids. As a result, ibuprofen and alcohol provoked synergistic hepatotoxicity against hepatocytes, and their toxicity increased prominently in 3D culture upon extended exposure. Oxidative stress appeared to be the mechanisms underlying the synergistic toxicity of ibuprofen and alcohol as evidenced by increased production of ROS and expression of the endogenous antioxidant system. Collectively, this study has demonstrated that ibuprofen and EtOH can induce synergistic hepatotoxicity, providing a line of evidence for caution against the use of ibuprofen in combination with alcohol.