• 생약학회지
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• 제30권2호
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• pp.222-225
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• 1999

The protective effects of the water extracts of nine kinds of medicinal herbs, which have been reputed to having the hepatoprotective activity in Chinese herbal medicine, on BNL cl.2 cells using a MTT assay were investigated. Five extracts including Coriolus versica, Curcuma longa, Phellinus linteus, Po-lygonum aviculare, and Salvia miltiorrhiza showed the protective effects on BNL cl.2 cells damaged by $CCl_4$ with $ED_{50}$ values of less than $100\;{\mu}g/ml$. Silymarin had been used as a positive control.

• 대한의생명과학회지
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• 제5권1호
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• pp.85-93
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• 1999

마우스 간 세포주인 BNL CL.2의 시험관내 배양에서 혈청과 IFN-$\gamma$가 세포주의 nitric oxide (NO)생성과 세포 손상에 미치는 영향을 알아보기 위한 실험을 하였다. 혈청이 공급된 배양에서 IFN-$\gamma$에 의한 세포 생존율은 거의 변동이 없었으나, 혈청을 제거한 배양에서는 약 65%의 생존율이 유지되었으며, NO생성 억제제인 N$^{G}$-monomethy-L-arginine (NMA)의 첨가는 농도 의존적으로 세포의 생존율을 감소시켰다. 혈청이 제거된 BNL CL.2 세포주는 IFN-$\gamma$ 단독 처리에서도 NO를 생성할수 있었으며, IFN-$\gamma$와 lipopolysaccharide (LPS)의 복합 처리는 세포주의 NO 생성을 상승적으로 증가시 켰다. 또한 protein tyrosine kinase (PTK) inhibitor인 herbimycin A와 genistein에 의해서 NO 생성이 억제되어 PTK의 활성이 혈청이 고갈된 BNL CL.2세포에서 NO의 생성에 중요한 역할을 담당하고 있기 때문으로 판단된다. IFN-$\gamma$의 독성은 혈청을 제거시킬 때 NO 생성 억제제에 상승적으로 간세포를 손상시키며, 이때 NO가 IFN-$\gamma$에 의해 유도된 손상을 어느 정도 억제시키는 것을 알 수 있었다.

• Natural Product Sciences
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• 제10권3호
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• pp.129-133
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• 2004

Quercetin is a dietary anticancer chemical that is capable of inducing apoptosis in tumor cells. However, little is known about its biological effect in nonmalignant hepatic cells. Using embryonic normal hepatic cell line (BNL CL.2) and its SV40-transformed tumorigenic cell line (BNL SV A.8), we evaluated the effects of quercetin on cell proliferation and apoptosis. As the results, our present study demonstrated that quercetin had a selective growth inhibition in normal versus tumorigenic hepatic cells such that BNL SV A.8 cells were very sensitive to the quercetin-mediated cytotoxicity. In particular, as evidenced by the increased number of positively stained cells in the TUNEL assay, the induction of characteristic nuclear DNA ladders, and the migration of many cells to sub-G1 phase in the BNL SV A.8 cells, quercetin treatment more sensitively induced apoptosis in BNL SV A8 cells than in BNL CL.2 cells. Collectively, our findings suggest that quercetin can be approached as a potential agent that is capable of inducing selective growth inhibition and apoptosis of hepatic cancer cells.

• Natural Product Sciences
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• 제10권6호
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• pp.296-301
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• 2004

Previously, a flavonoid fraction, which consisted mainly of protocatechuic acid, fustin, fisetin, sulfuretin, and butein, here named RCMF $({\underline{R}}VS\;{\underline{c}}hloroform-{\underline{m}}ethanol\;{\underline{f}}raction)$, was prepared from a crude acetone extract of Rhus verniciflua Stokes (RVS) which is traditionally used as a food additive and as an herbal medicine. In this study, we evaluated the effects of RCMF on the antioxidant defense system using embryonic normal hepatic cell line (BNL CL.2) and its SV40-mediated transformed cell line (BNL SV A.8). This study demonstrates that RCMF selectively stimulated the antioxidant defense system of normal cells, as BNL CL.2 cells proved to be more sensitive to RCMF-mediated increases of superoxide dismutase, catalase, glutathione, and glutathione reductase than BNL SV A.8 cells. In particular, RCMF caused a significant increase in the malonaldehyde content of BNL SV A.8 cells, which is believed to be closely associated with cytotoxicity of RCMF and RCMF-mediated growth inhibition. Collectively, our findings suggest that the flavonoid fraction, RCMF, selectively stimulates the antioxidant defense system in normal rather than hepatic tumor cells.

• Molecular & Cellular Toxicology
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• 제5권1호
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• pp.32-43
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• 2009

Acetaminophen (APAP) overdose is known to cause severe hepatotoxicity mainly through the depletion of glutathione. In this study, we compared the cytotoxic effects of APAP on both a normal murine hepatic cell line, BNL CL.2, and its SV40-transformed cell line, BNL SV A.8. Gene expression profiles for APAP-treated cells were also obtained using microarray and analyzed to identify differences in genes or profiles that may explain the differences of susceptibility to APAP in these cell lines. These two cell lines exhibited different susceptibilities to APAP (0-$5,000{\mu}M$); BNL SV A.8 cells were more susceptible to APAP treatment compared to BNL CL.2 cells. A dose of $625{\mu}M$ APAP, which produced significant differences in cytotoxicity in these cell lines, was tested. Microarray analysis was performed to identify significant differentially expressed genes (DEGs) irrespective of APAP treatment. Genes up-regulated in BNL SV A.8 cells were associated with immune response, defense response, and apoptosis, while down-regulated genes were associated with catalytic activity, cell adhesion and the cytochrome P450 family. Consistent with the cytotoxicity data, no significant DEGs were found in BNL CL.2 cells after treatment with $625{\mu}M$ APAP, while cell cycle arrest and apoptosis-related genes were up-regulated in BNL SV A.8 cells. Based on the significant fold-changes in their expression, a genes were selected and their expressions were confirmed by quantitative real-time RT-PCR; there was a high correlation between them. These results suggest that gene expression profiles may provide a useful method for evaluating drug sensitivity of cell lines and eliciting the underlying molecular mechanism. We further compared the genes identified from our current in vitro studies to the genes previously identified in our lab as regulated by APAP in both C57BL/6 and ICR mice in vivo. We found that a few genes are regulated in a similar pattern both in vivo and in vitro. These genes might be useful to develop as in vitro biomarkers for predicting in vivo hepatotoxicity. Based on our results, we suggest that gene expression profiles may provide useful information for elucidating the underlying molecular mechanisms of drug susceptibility and for evaluating drug sensitivity in vitro for extrapolation to in vivo.

• Toxicological Research
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• 제30권4호
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• pp.261-266
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• 2014

Environmental pollutants such as polycyclic aromatic hydrocarbons (PAHs) have been implicated in cancer development and progression. However, the effects of PAHs on carcinogenesis are still poorly understood. Here, we characterized a mouse cancer cell line BNL 1ME A. 7R.1 (1MEA) derived by transformation of non-tumorigenic liver cell line BNL CL.2 (BNL) using 3-methylcholanthrene (3MC), a carcinogenic PAH. RT-PCR and immunoblot analysis were used to determine the expression level of mRNA and proteins, respectively. To determine functionality, cell motility was assessed in vitro using a transwell migration assay. Both mRNA and protein levels of E-cadherin were significantly decreased in 1MEA cells in comparison with BNL cells. While the expression levels of mesenchymal markers and related transcription factors were enhanced in 1MEA cells, which could lead to increase in cell motility. Indeed, we found that 7-day exposure of BNL cells to 3-MC reduced the level of the adhesion molecule and epithelial marker E-cadherin and increased reciprocally the level of the mesenchymal marker vimentin in a dose-dependent manner. Taken together, these results indicate that the process of epithelial-mesenchymal transition (EMT) may be activated during premalignant transformation induced by 3-MC. A mechanism study to elucidate the relation between 3-MC exposure and EMT is underway in our laboratory.

• Nutrition Research and Practice
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• 제1권3호
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• pp.180-183
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• 2007

Cytoprotective ability of polysaccharides isolated from different edible mushrooms was investigated on the 7-ketocholesterol-induced damaged cell line. Polysaccharide extracts from six different edible mushrooms-Flammulina velutipes, Peurotus ostreatus, Lentinus edodes, Agrocybe aegerita, Agaricus blazei, and Cordyceps militaris- were prepared by hot water extraction and alcohol precipitation. Cytoprotective ability was evaluated by measuring the viable cells of the normal embryonic liver cell line (BNL CL. 2) in the presence of 7-ketocholesterol. At $80\;{\mu}g/mL$ of 7-ketocholesterol, cytotoxicity was very high with a loss of 98% of viable cells after 20 h of incubation. With the addition of $200\;{\mu}g/mL$ of each polysaccharide isolate to the cell line containing $80\;{\mu}g/mL$ of 7-ketocholesterol, polysaccharide isolates from both Flammulina velutipes and Peurotus ostreatus could significantly inhibit the 7-ketochoelsterol-induced cytotoxicity in the cells. But other polysaccharide isolates were not effective in inhibiting cell damage caused by the oxLDL-induced cytotoxicity.

• Journal of Microbiology and Biotechnology
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• 제25권6호
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• pp.795-802
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• 2015

This study investigates the bioactivity of tannin from amaranth (Amaranthus caudatus L.) extracts. The antioxidant activities of the extracts from amaranth leaves, flowers, and seeds were evaluated. Tannin from leaves of amaranth has been evaluated for superoxide scavenging activity by using DPPH and ABTS⁺ analysis, reducing power, protective effect against H₂O₂-induced oxidative damage in L-132 and BNL-CL2 cells, and inhibition of superoxide radical effects on HL-60 cells. At a concentration of 100 µg/ml, tannin showed protective effects and restored cell survival to 69.2% and 41.8% for L-132 and BNL-CL2 cells, respectively. Furthermore, at the same concentration, tannin inhibited 41% of the activity of the superoxide radical on HL-60 cells and 43.4% of the increase in nitric oxide levels in RAW 264.7 cells. The expression levels of the antioxidant-associated protein SOD-1 were significantly increased in a concentration-dependent manner in RAW 264.7 cells treated with tannin from amaranth leaves. These results suggest that tannin from the leaves of Amaranthus caudatus L. is a promising source of antioxidant component that can be used as a food preservative or nutraceutical.

• 생명과학회지
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• 제27권4호
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• pp.442-449
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• 2017

엉겅퀴(C. japonicum) 부위별 추출물을 기능성 식품소재로 활용가능성을 알아보기 위해서 총 페놀 및 silymarin 화합물의 함량분석, 항산화 및 간 보호효과를 서양엉겅퀴와 비교하였다. 엉겅퀴의 총 페놀함량은 지상부($97.22{\pm}5.51mg/g$)가 지하부($85.32{\pm}3.06mg/g$)보다 많았으며, 서양엉겅퀴의 전초보다 다소 낮은 함량을 보였다. 엉겅퀴의 silymarin 화합물의 총 함량은 서양엉겅퀴의 55.56%이었으며, 지하부($0.47{\pm}0.03mg/g$)는 지상부($0.18{\pm}0.02mg/g$)보다 많이 함유되어 있었다. 또한 지상부에서는 silychristin, silydianin이, 지하부에서는 silychristin, silydianin, silybin B, isosilybin B 등이 검출되었다. 엉겅퀴의 항산화활성은 대체로 서양엉겅퀴보다는 약간 낮았고, 지하부가 지상부보다 높은 활성을 보였다. 엉겅퀴 추출물을 1 mg/ml 농도로 처리하였을 때, DPPH 활성은 지상부와 지하부에서 각각 $83.76{\pm}0.60%$, $88.28{\pm}0.17%$의 활성을 나타났으며, FRAP 활성은 지상부와 지하부에서 각각 $77.63{\pm}0.70$, $82.83{\pm}0.39%$로 나타났다. ABTS 활성도 엉겅퀴 추출물을 0.1 mg/ml 농도로 처리하였을 때, 지하부와 지상부에서 각각 $68.60{\pm}1.24%$와 $63.41{\pm}0.57%$로 나타났다. 엉겅퀴 추출물의 간 보호효과는 지하부에서 지상부보다 다소 높은 활성이 나타났으나 서양엉겅퀴 지상부의 활성보다는 낮았다. t-BHP, $H_2O_2$ 및 Ethanol의 처리에 따른 손상된 간세포에 엉겅퀴 추출물을 0.2 mg/ml씩 처리하였을 때, 세포생존율은 각각 $49.58{\pm}0.34$, $76.87{\pm}1.10$ 및 $71.73{\pm}0.58%$로서 추출물을 처리하지 않았을 때보다 각각 24.78, 61.32 및 38.04%씩 높아졌다. 이와 같은 결과로 볼 때, 본 연구에서는 국내산 엉겅퀴의 간 보호효과를 활용한 기능성 식품의 개발이 가능할 것으로 생각되었다.

• 생명과학회지
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• 제27권8호
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• pp.909-918
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• 2017

본 연구는 아이스플랜트의 생리활성을 향상시키고, 식생활에서의 활용가능성을 높이기 위하여 물김치 형태로 발효시켰으며, 아이스플랜트의 성분 및 몇 가지 생리활성을 발효 전과 비교하였다. 물김치 침지액의 pH는 발효 6일째까지 계속해서 낮아졌으며, lactic acid의 생성량은 발효 4일째에 가장 많았다. 아이스플랜트 물김치 건더기 중의 D-pinitol은 발효 전 $4.196{\pm}0.096mg/g$에서 6일째에는 $3.843{\pm}0.116mg/g$으로 감소하였다. 그러나 myo-inositol과 D-chiro-inositol은 각각 발효 전 $0.453{\pm}0.022$와 $2.815{\pm}0.008mg/g$에서 6일째에 $0.630{\pm}0.029$와 $4.117{\pm}0.096mg/g$으로 증가하였다. 아이스플랜트 추출물의 DPPH, ABTS radical 소거활성 및 FRAP 활성은 대체로 발효 후에 높게 나타나 유사한 경향이었으며, 1 mg/ml의 농도로 처리하였을 때 각각 발효 5일째($79.09{\pm}0.69%$), 4일째($87.55{\pm}1.21%$) 및 6일째($78.72{\pm}0.99%$)에 가장 높았다. Lipid/MA assay에 의한 항산화 활성도 대체로 발효 후에 높게 나타나 유사한 경향이었으며, 추출물을 $500{\mu}g/ml$ 처리하였을 때, 발효 4일째에 가장 높은 활성($64.65{\pm}1.63%$)을 나타났으며, 발효 5, 6일째에는 낮아지는 경향이었다. 아이스플랜트 추출물의 t-BHP, $H_2O_2$ 및 ethanol에 의하여 손상된 BNL CL.2 세포의 생존율은 각각 $14.19{\pm}0.98$, $13.80{\pm}2.25$ 및 $25.89{\pm}2.90%$이었으나, 추출물을 $200{\mu}g/ml$의 농도로 처리하였을 경우, 각각 발효 6일째에 $50.07{\pm}4.85%$, 2일째에 57.06% 및 4일째에 $66.06{\pm}1.36%$의 세포생존율을 나타내어 간세포가 회복되고 있음을 알 수 있었다. 그러나 발효 후에 간세포 보호효과가 뚜렷이 증가하지는 않았다. ${\alpha}-Glucosidase$ 저해활성은 모든 실험군에서 $83.52{\pm}2.69{\sim}92.79{\pm}2.16%$로 상당히 높게 나타났으며, 발효기간이 경과함에 따라 조금씩 증가하였다. 한편 ${\alpha}-Amylase$ 저해활성은 전체적으로 $52.64{\pm}3.26{\sim}57.56{\pm}1.21%$로 나타났으며 발효 전후에 뚜렷한 차이가 없었다. 이와 같은 결과로 볼 때, 아이스플랜트의 물김치 발효기간은 4~5일이 적정할 것으로 생각되었다. 한편 발효에 의한 기능성은 항산화효과를 제외한 간세포보호 및 ${\alpha}-Glucosidase$, ${\alpha}-Amylase$ 저해활성은 전체적으로 소폭 향상에 그쳤다. 따라서 이러한 결과를 바탕으로 발효 등 다양한 가공방법이 개발되어, 향후 아이스플랜트의 소비촉진의 계기가 되기를 기대한다.