• 한국어류학회지
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• 제12권3호
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• pp.180-185
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• 2000

플라스틱의 원재료로 이용되는 bisphenol-A (BPA)가 vitellogenin (VTG)의 합성에 미치는 영향을 넙치의 간세포를 초대 배양하여 조사하였다. 간세포는 2일간 사전 배양을 하였으며, estradiol-$17\beta(E_2)$ 및 BPA를 배양액에 동시 첨가하였다. 간세포는 6일간 배양한 후 배양액을 회수하여 SDS-PAGE로 분석하였다. BPA는 농도 의존적으로 총 단백질에 대한 VTG 비율은 증가하였으며, $10^{-6}M$과 $10^{-5}M$의 농도에서 유의한 차이를 나타내었다(P<0.05). 특히, BPA $10^{-5}M$의 농도에서의 총 단백질에 대한 VTG 비율은 26.36%로 $E_2$만을 첨가한 대조군과 유의한 차이를 나타내지 않았다. $E_2$와 BPA를 단독 또는 동시에 첨가하여 유도된 VTG합성은 배양액에 tamoxifen$(10^{-6}M)$을 첨가함으로서 $E_2$를 첨가한 대조군의 약 80%로 유의하게 억제되었다. 그리고, in vivo에서 $E_2$에 의해 유도된 VTG합성은 in vitro에서 간세포를 6일간 배양함에 따라 $E_2$를 첨가한 대조군의 약 22%까지 억제되었다. 이러한 감소효과는 BPA의 농도 의존적으로 지연되는 결과를 나타내었다. 따라서 넙치에서의 BPA는 $E_2$ receptor와의 반응을 통한 에스트로겐 작용에 의해 VTG합성을 유도하고, 난황형성기에서의 VTG 합성을 장시간 지속시킬 가능성이 있는 것으로 추측된다.

• 한국자원식물학회지
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• 제19권6호
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• pp.649-654
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• 2006

The objective of present study was to investigate the anti oxidative and hepatoprotective effects of tomato extracts. Total antioxidant capacity and total antioxidant response were 5.5 and $19.8{\mu}g$ Trolox equivalent per mg of tomato extract, respectively. DPPH radical scavenging activity of tomato extracts ($10mg\;ml^{-1}$) was 70% as compared to 100% by pyrogallol solution as a reference. The effect of the tomato extracts on lipid peroxidation was examined using rat liver mitochondria induced by iron/ascorbate. Tomato extracts at the concentration of $0.5mg\;ml^{-1}$ significantly decreased TBARS concentration. Tomato extracts prevented lipid peroxidation in a dose-dependent manner. The effect of the tomato extracts on reactive oxygen species (ROS) generation was examined using cell-free system induced by $H_2O_2/FeSO_4$. Addition of $1mg\;ml^{-1}$ of tomato extracts significantly reduced dichlorofluorescein (DCF) fluorescence. Tomato extracts caused concentration-dependent attenuation of the increase in DCF fluorescence, indicating that tomato extracts significantly prevented ROS generation in vitro. The effect of tomato extracts on cell viability and proliferation was examined using hepatocyte culture. Primary cultures of rat hepatocytes were incubated with 1mM tert-butyl hydroperoxide (t-BHP) for 90 min in the presence or absence of tomato extracts. MTT values by addition of tomato extracts at the concentration of 2, 10, and $20mg\;ml^{-1}$ in the presence of t-BHP were 13, 33 and 48%, respectively, compared to 100% as control. Tomato extracts increased cell viability in a dose-dependent manner. These results demonstrate that tomato extracts suppressed lipid peroxidation and t-BHP-induced hepatotoxicity and scavenged ROS generation. Thus antioxidant and hepatoprotective effects of tomato extracts seem to be due to, at least in part, the prevention from free radicals-induced oxidation, followed by inhibition of lipid peroxidation.

• 동아시아식생활학회지
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• 제4권2호
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• pp.59-67
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• 1994

The present study was conducted to evaluate the hepatoprotective effect of puerariae Radix methanol extract on benzo(a) pyrene(B(a)P) - induced liver injuries in rats. In vitro experiment, primary cultured hepatocytes (5X105 cells/$m\ell$) were cultured for 20~24 hours after adding puerariae Radix mehtanol extract(32$\mu\textrm{g}$/$m\ell$) and B(a)P(50 uM). In vivo experiment, Puerariae Radix methanol extract(0.25 g/kg/day, per os) was administered for 7 days and B(a)P(0.1 mg/kg/day, intraperitoneally) was given after the last administration of extract. And then the hepatoprotective effect of Puerariae Radix methanol extract was investigated biochemically through in vitro and in vivo experiments. Namely, activities of enzymes (GOT, GPT and LDH) were measured and 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide(MTT) assay were carried out in vitro cell culture study and GOT, GPT, LDH and ALP activities and HDL-cholesterol, total cholesterol and triglyceride contents were performed in vivo study. In vitro experiment, as a result of enzyme activity measurement(GOT, GPT and LDH) and MTT assay, GOT,GPT and LDH activities changed by B(a)P were recovered to normal levels and hepatocytes impaired by B(a)P were recovered to normal. In vivo experiment, Puerariae Radix methanol extract significantly decreased the enzyme activities(GOT, GPT, ALP and LDH in serum and GPT and ALP in tissue) and lipid contents in comparison to B(a)P-treated group.

• Animal Bioscience
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• 제34권2호
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• pp.312-319
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• 2021

Objective: Stress-induced cytotoxicity caused by xenobiotics and endogenous metabolites induces the production of reactive oxygen species and often results in damage to cellular components such as DNA, proteins, and lipids. The cytochrome P450 (CYP) family of enzymes are most abundant in hepatocytes, where they play key roles in regulating cellular stress responses. We aimed to determine the effects of the antioxidant compound, methylsulfonylmethane (MSM), on oxidative stress response, and study the cytochrome P450 family 3 subfamily A (CYP3A) gene expression in fetal horse hepatocytes. Methods: The expression of hepatocyte markers and CYP3A family genes (CYP3A89, CYP3A93, CYP3A94, CYP3A95, CYP3A96, and CYP3A97) were assessed in different organ tissues of the horse and fetal horse liver-derived cells (FHLCs) using quantitative reverse transcription polymerase chain reaction. To elucidate the antioxidant effects of MSM on FHLCs, cell viability, levels of oxidative markers, and gene expression of CYP3A were investigated in H2O2-induced oxidative stress in the presence and absence of MSM. Results: FHLCs exhibited features of liver cells and simultaneously maintained the typical genetic characteristics of normal liver tissue; however, the expression profiles of some liver markers and CYP3A genes, except that of CYP3A93, were different. The expression of CYP3A93 specifically increased after the addition of H2O2 to the culture medium. MSM treatment reduced oxidative stress as well as the expression of CYP3A93 and heme oxygenase 1, an oxidative marker in FHLCs. Conclusion: MSM could reduce oxidative stress and hepatotoxicity in FHLCs by altering CYP3A93 expression and related signaling pathways.

• 한국간담췌외과학회지
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• 제27권4호
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• pp.342-349
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• 2023

Backgrounds/Aims: Liver organoids have emerged as a powerful tool for studying liver biology and disease and for developing new therapies and regenerative medicine approaches. For organoid culture, Matrigel, a type of extracellular matrix, is the most commonly used material. However, Matrigel cannot be used for clinical applications due to the presence of unknown proteins that can cause immune rejection, batch-to-batch variability, and angiogenesis. Methods: To obtain human primary hepatocytes (hPHs), we performed 2 steps collagenase liver perfusion protocol. We treated three small molecules cocktails (A83-01, CHIR99021, and HGF) for reprogramming the hPHs into human chemically derived hepatic progenitors (hCdHs) and used hCdHs to generate liver organoids. Results: In this study, we report the generation of liver organoids in a collagen scaffold using hCdHs. In comparison with adult liver (or primary hepatocyte)-derived organoids with collagen scaffold (hALO_C), hCdH-derived organoids in a collagen scaffold (hCdHO_C) showed a 10-fold increase in organoid generation efficiency with higher expression of liver- or liver progenitor-specific markers. Moreover, we demonstrated that hCdHO_C could differentiate into hepatic organoids (hCdHO_C_DM), indicating the potential of these organoids as a platform for drug screening. Conclusions: Overall, our study highlights the potential of hCdHO_C as a tool for liver research and presents a new approach for generating liver organoids using hCdHs with a collagen scaffold.

• 대한기계학회:학술대회논문집
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• 대한기계학회 2008년도 추계학술대회A
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• pp.1419-1426
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• 2008

Fulminant hepatic failure is a clinical syndrome associated with a high mortality rate. Orthotopic liver transplantation is the only clinically proven effective treatment for patients with end-stage liver disease who do not respond to medical management. A major limitation of this treatment modality is the scarcity of donor organs available, resulting in patients dying while waiting for a donor liver. An extracorporeal bioartificial liver (BAL) device containing viable hepatocytes has the potential to provide temporary hepatic support to liver failure patients, serving as a bridge to transplantation while awaiting a suitable donor. In some patients, providing temporary hepatic support may be sufficient to allow adequate regeneration of the host liver, thereby eliminating the need for a liver transplant. Although the BAL device is a promising technology for the treatment of liver failure, there are several technical challenges that must be overcome in order to develop systems with sufficient processing capacity and of manageable size. In this overview, the authors describe the critical issues involved in developing a BAL device. They also discuss their experiences in hepatocyte culture optimization within the context of a microchannel flat-plate BAL device.

• 한국축산식품학회지
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• 제18권4호
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• pp.348-357
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• 1998

This study was conducted to investigate the effects of mugwort extracts on the blood ethanol concentration and liver function in rats. Sprague-Dawley rats were used, the rats administered with 25% ethanol (5g/kg$.$B.W.) were devided into three groups (CON-E ; 25% ethanol administered to the CON-E) according to the administered ethanol concentration and the levels of administered mugwonts. Mugwont roots extracts were administered via the caudal vein. Ethanol concentration was measured at the time of 0, 1, 2 and 3hr by gas chromatography. GOT(Glutamic Oxaloacetic Transaminase) and GPT(Glutamic Pyruvic Transaminase) were measured at the time of 0 and 5hr. Components of each extracts were analyzed by using high performance liquid chromatography. Cell number, GOT and GPT were investigated by using rat hepatocyte culture. Megwort extracts were added at the levels of 1% or 2%. Hepatocyte culture were into five groups according to the addition levels. The results were summarized as follows ; 1. Catechin contents of 8∼10mg/100g and the contents of (-)-epigallocatechin was high in the water extracts. 2. Ethanol degradation efficiency declines in the following order : MDW-E＞MOH-E＞CON-E. 3. The numbers of rat hepatocytes declines in the following order : 2% MDW-L＞1%MDW-L＞1%MOH-L＞CON-L＞2%MOH-L. These results suggest that crude catechin of mugwort extracts may play important roles to degrade ethanol and recover liver function in rats.

• 한국양식학회지
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• 제15권1호
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• pp.31-37
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• 2002

이 연구는 무지개송어, O. mykiss 간세포 배양을 이용해 합성화학물질인 2,4-Dichlorophenoxy acetic acid (2,4-D)가 vitellogenin (VTG) 합성과 estrogen (E$_2$) - estrogen receptor (ER) binding affinity에 미치는 영향을 조사하였다. 간세포는 2일간 전 배양한 후에 E, (2$\times$10$\^$-6/ M) 또는 2,4-D (10$\^$-9/∼10$\^$-6/ M)를 배지에 첨가해 5일간 배양한 후, VTG과 E$_2$-ER binding affinity를 SDS-PAGE와 ELISA를 이용해 각각 조사하였다. 사용된 2,4-D의 농도는 세포의 형태, 생존력 및 DNA 함량에는 영향을 미치지 않았다. 또한, 2,4-D는 VTG의 합성에도 어떠한 영향도 미치지 않았으나 E$_2$-ER binding affinity를 억제하였다. 이 binding affinity는 2,4-D의 농도가 증가할수록 증가하는 경향을 나타내어 10$\^$-7/과 10$\^$-6/ M에서 25%와 30%를 각각 억제하였다. 이 결과들은 비록 2,4-D가 VTG의 합성에는 영향을 미치지 않지만, ER에서 xeno-estrogenic contaminant로서 행동하는 것을 제시하였다.

• Toxicological Research
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• 제22권1호
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• pp.23-30
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• 2006

Among poly aromatic hydrocarbons, dioxin and PCBs are the most controversial environmental pollutants in our modern life. These pollutants are known as human carcinogens, and liver is the most sensitive target in animal cancer models. Specific aims of the study were focused on the mechanism of carcinogenesis in hepatocytes and the structure-activity relation among these diverse environmental chemicals. Because key mechanisms of dioxin-induced carcinogenesis in human epithelial cell model are the alteration of signal transduction pathway and PKC isoforms, the alteration of the signal transduction pathways and other factors associated with carcinogenesis were studied. Rat hepatocytes cultured under the sandwich protocols were exposed with the various concentration of dioxins and PCBs, and signal transduction pathway, protein kinase C isoforms, oxidant stress, and apoptotic nuclei were evaluated. Since it is important to understand the structure-activity relation among these chemicals to properly assess the carcinogenic potentials, the study analyzed the parameters associated with carcinogenic processes, based on their structural characteristics. In addition, signal transduction pathways and PKC isoforms involved in inhibition of UV-induced apoptosis were also analyzed to elaborate the tumor promotion mechanism of these chemicals. Induction of apoptosis by UV irradiation was optimal at $60\;J/m^2$ in primary hepatocyte in culture. Compared to non coplanar PCBs such as PCB 114 and PCB 153, coplanar PCBs such as PCB 77 and PCB126 showed a stronger inhibition of apoptosis induced by UV irradiation. Production of reactive oxygen species (ROS) was more stimulated by non-coplanar PCBs than coplanar PCBs with the most potent induction of ROS by chlorinated non-coplanar PCB. As compared to the level of induction by PCB126, non-coplanar PCB153 showed a higher increase of intracellular concentrations. Besides the alteration of intracellular calcium concentration, translocation of PKC from cytosolic fraction to membrane fraction was clearly observed upon the exposure of non-coplanar PCB. Taken together, the present study demonstrated that there is a potent structure-activity relationship among PCB congeners and the mechanism of PAH-induced carcinogenesis is structure-specific. The study suggested that more diverse pathways of PAH-induced carcinogenesis should be taken into account beyond the boundary of Ah receptor dogma to assess the health impact of PAH with more accuracy.

• 한국식품위생안전성학회지
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• 제28권2호
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• pp.89-94
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• 2013

마늘 숙성 중 생성되는 S-allylmercaptocysteine의 콜레스테롤 생합성 억제 효과에 대하여 Hepatocytes를 이용하여 조사하였다. HepG2 cells을 Dulbecco's modified Eagle's medium (DMEM)에 배양하여 S-allylmercaptocysteine의 농도를 20, 40, 60, 80 및 100 mL 씩 각각 첨가하여 cell viability를 살펴본 결과 20~40 ${\mu}g/mL$에서는 높았으며, 60 ${\mu}g/mL$ 농도에서 약 50%가 유지되었다. S-allylmercaptocysteine을 5, 10, 15 및 20 ${\mu}g/mL$ 농도로 [$^{14}C$]-acetatecholesterol에서 처리하였을 경우 15 ${\mu}g/mL$ 농도에서 cholesterol 생합성이 79%로 억제되었다. Fatty acid synthase의 활성은 0.95 nmol에서 19%의 억제효과를 나타내었으나, Glucose 6-phosphate dehydrogenase (G6PDH)의 활성에는 거의 영향을 미치지 않았다. S-allylmercaptocysteine의 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase의 활성은 용량 의존형으로 감소하였다. 이상의 결과로 보아 마늘 숙성 과정에서 생성되는 주요 성분인 S-allylmercaptocysteine은 간 세포에서 cholesterol의 생합성을 억제하는데 기여하는 것으로 나타났다.