• Korean Journal of Pharmacognosy
• /
• v.30 no.2
• /
• pp.222-225
• /
• 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.

• Biomedical Science Letters
• /
• v.5 no.1
• /
• pp.85-93
• /
• 1999

Nitric oxide (NO) plays an important role in immunologic defense, and influences upon the functioning of secretory tissues and cells. It also exhibits cytotoxic/cytostatic activity as one of major operating effectors of the cellular immunity system. We investigated the effects of serum on the cell damages and NO production in the mouse liver cell line BNL CL.2 to establish the role of NO. We observed that, when BNL CL.2 cells were cultured in serum-free medium, they were induced to cell damage by the stimulation of IFN-$\gamma$ alone or IFN-$\gamma$ plus LPS. Serum-starved cells showed large amount of nitrite accumulation and NO synthase (NOS) expression in response to IFN-$\gamma$ alone in dose- and time- dependent manners, but serum-supplied cells did not The production of NO was blocked by protein tyrosine kinase (PTK) inhibitors, genistein and herbimycin. These results suggest that the deprivation of serum in the BNL CL.2 cell culture medium might primed with the cells to produce NO when the cells are triggered by IFN-$\gamma$ and the involvement of PTK signal transduction pathway in the expression of NOS gene in murine hepatocytes.

• Natural Product Sciences
• /
• v.10 no.3
• /
• pp.129-133
• /
• 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
• /
• v.10 no.6
• /
• pp.296-301
• /
• 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
• /
• v.5 no.1
• /
• pp.32-43
• /
• 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
• /
• v.30 no.4
• /
• pp.261-266
• /
• 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
• /
• v.1 no.3
• /
• pp.180-183
• /
• 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
• /
• v.25 no.6
• /
• pp.795-802
• /
• 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.

• Journal of Life Science
• /
• v.27 no.4
• /
• pp.442-449
• /
• 2017

We analyzed the content of total phenolic and silymarin compounds of Cirsium japonicum (CJ), and its antioxidant activities and Liver protective effects were compared with those of Silybum marianum (SM). The total phenolic content in the aerial part ($97.22{\pm}5.51mg/g$) of CJ is higher than that in the underground part ($85.32{\pm}3.06mg/g$). The total silymarin content of CJ was 55.56% of SM, with the underground part ($0.47{\pm}0.03mg/g$) having higher content than the aerial part ($0.18{\pm}0.02mg/g$). The antioxidant activity of CJ was generally slightly lower than that of milk thistle, and the underground part of CJ generally had higher activity compared to the aerial part. When CJ extracts were processed at 1 mg/ml, DPPH activities were $83.76{\pm}0.60$ and $88.28{\pm}0.17%$, and FRAP activities were $77.63{\pm}0.70$ and $82.83{\pm}0.39%$ for extracts from aerial part and underground part, respectively. ABTS activities were $68.60{\pm}1.24$ and $63.41{\pm}0.57%$ for underground and aerial part respectively when extracts were processed at 0.1 mg/ml. The Liver protective effects of CJ were higher in the extracts from underground part compared to the aerial part, Liver cells were damaged by treating them with t-BHP, $H_2O_2$ and Ethanol, and then they were treated with 0.2 mg/ml CJ extracts. The survival rates of the damaged liver cells were $49.58{\pm}0.34$, $76.87{\pm}1.10$ and $71.73{\pm}0.58%$ respectively, which were higher than the cells not treated with extracts.

• Journal of Life Science
• /
• v.27 no.8
• /
• pp.909-918
• /
• 2017

Ice plant (Mesembryanthemum crystallinum L.) was fermented in brine in the form of mulkimchi (IPMB), and its contents of organic acid and cyclitols and biological activities were compared with those before fermentation. The pH of the IPMB continuously decreased until the sixth day of fermentation. The lactic acid yield was greatest on the fourth day. D-pinitol in ice plant mulkimchi solids (IPMS) decreased during fermentation. However, myo-inositol and D-chiro-inositol increased. The radical scavenging activities of ABTS and DPPH, in addition to the activity of FRAP, of the IPMS extract were generally higher after fermentation, with the activities highest on the fifth ($79.09{\pm}0.69%$), fourth ($87.55{\pm}1.21%$), and sixth ($78.72{\pm}0.99%$) days of fermentation, respectively, when treated with 1 mg/ml of the extract. As shown by a lipid/MA assay, antioxidant activity was generally higher after fermentation. The viability of BNL CL.2 cells damaged by t-BHP, $H_2O_2$, and ethanol was $14.19{\pm}0.98$, $13.80{\pm}2.25$, and $25.89{\pm}2.90%$, respectively. When treated with $200{\mu}g/ml$ of IPMS extract, the cell viability was $57.06{\pm}4.52%$ on the first day, and $66.06{\pm}1.36%$ on the fourth day, and $50.07{\pm}04.85%$ on the sixth day of fermentation. Hepatocyte protective effects did not increase significantly after fermentation. ${\alpha}-glucosidase$ inhibitory activity was quite high, with a range of $83.52{\pm}2.69$ to $92.79{\pm}2.16%$, and the activity increased gradually in all the groups over the fermentation period. There was no clear correlation between ${\alpha}-amylase$ inhibitory activity and fermentation.