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 verni...
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.
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.
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문제 정의
This study demonstrates that RCMF selectively triggers the stimulation of the antioxidant defense system in normal hepatic cells. However, further detailed in vivo and in vitro studies are needed to determine the details of the mechanism involved in the RCMF-mediated selective stimulation of the antioxidant defense system in hepatocytes.
가설 설정
1). Based on these results, we hypothesized that the significant increase in the MDA content in the BNL SV A.8 cells is closely related to the RCMF-mediated cytotoxicity in the cells.
제안 방법
To evaluate the hypothesis, we examined the growth inhibitory response to RCMF by determining the level of tritium incorporated by BNL CL.2 or BNL SV A.8 cells (Fig. 2). Tritim uptake assay revealed that RCMF treatment actively inhibited tritium incorporation by BNL SV A.
이론/모형
GR activity was determined by measuring the decrease in the NADPH concentration, and is expressed as nmol of NADPH/min/mg protein. Cellular GSH content was determined using the method described by Dringen and Hamprecht (1996).
Determination of cytotoxicity - Cellular cytotoxicity induced by RCMF treatment was determined by trypan blue exclusion assay. Briefly, the hepatic cells were cultured in DMEM supplemented with 10% FBS inpresence of 100)ig/ml RCMF for 12 h and 24 h.
후속연구
This study demonstrates that RCMF selectively triggers the stimulation of the antioxidant defense system in normal hepatic cells. However, further detailed in vivo and in vitro studies are needed to determine the details of the mechanism involved in the RCMF-mediated selective stimulation of the antioxidant defense system in hepatocytes.
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