ABSTRACT
<p><b>OBJECTIVE</b>There are several reports of cellular-aging-dependent alterations in the antioxidant capacity of human fibroblasts. Fibroblasts show slower the growth rate at late passages (referred to hereafter as old cells) than at early passages (referred to hereafter as young cells). Antioxidants may control cellular growth by modulating reactive oxygen species (ROS). Methanolic extracts from broad beans (MEBB) contain phenolic compounds and have ROS-scavenging activities. In this study, we investigated the effects of MEBB on cellular growth and antioxidant levels in normal human lung fibroblasts.</p><p><b>METHODS</b>To determine cytosolic superoxide dismutase (SOD) activities, cytosolic glutathione peroxidase (GSH-Px) activities, catalase activities, reduced glutathione (GSH) concentrations, and growth rate, MEBB treatments were performed on young and old cells.</p><p><b>RESULTS</b>In young and old cells treated with 120 μg/ml MEBB, the growth rates increased by 28.1 and 15.2%, respectively, compared with controls. The MEBB treatment of young cells caused a 62.5% increase in SOD activity, but the treatment of old cells caused a 39.5% decrease. The catalase activities of the young and old cells treated with MEBB were equal to those of control cells. Young and old cells treated with MEBB were equal to the control cells in terms of GSH-Px activity. The GSH concentrations in the young and old cells treated with 120 μg/ml MEBB increased by 22.1 and 45.9%, respectively.</p><p><b>CONCLUSION</b>These studies elucidated a new cellular growth mechanism whereby human lung fibroblasts modulate intracellular GSH levels via the action of MEBB.</p>
ABSTRACT
<p><b>OBJECTIVE</b>Human diploid cells are more susceptible to oxidative stress at late passage than at early passage, presumably because of the decrease in cellular-reduced glutathione (GSH) concentration. Water-soluble protein (WSP) from broad beans scavenges free radicals. The effects of WSP on the glutathione system were examined in PDL 20 (early passage) and PDL 50 (late passage) human lung fibroblasts (TIG-1).</p><p><b>METHODS</b>To determine cytosolic glutathione peroxidase (GSH-Px) activities, glutathione reductase (GR) activities, oxidized glutathione (GSSG) concentrations, and GSSG/reduced glutathione (GSH) ratios, WSP and hydrocortisone (HC) treatments of TIG-1 cells (PDL 20→50 and PDL 50→75) were performed for 40 days. We also investigated the GSSG concentrations and GR activities in PDL 20 cells that were continuously treated with WSP until PDL 39 and 55.</p><p><b>RESULTS</b>GSSG concentrations decreased in WSP- and HC-treated PDL 50→75 cells. The GSSG/GSH ratios in PDL 50→75 cells became low after the treatments. Increases in GR activities were observed in treated PDL 50→75 cells. The decline in the GSSG concentration of PDL 50→75 cells correlated with the increase in GR activity. The GSSG levels in control cells were higher following cellular age, whereas the levels in treated cells were lower than those in the control. The studies on cellular age-related changes indicated that greater increases in GR activity were found in treated cells than in the control.</p><p><b>CONCLUSION</b>These results indicated that WSP influences the GSSG concentration that is associated with cellular aging, but the mechanism of GSSG reduction by WSP remains unknown. The enhancement of glutathione status following WSP treatment may be related to the delay in the cellular aging.</p>
ABSTRACT
Objective: Human diploid cells are more susceptible to oxidative stress at late passage than at early passage, presumably because of the decrease in cellular-reduced glutathione (GSH) concentration. Water-soluble protein (WSP) from broad beans scavenges free radicals. The effects of WSP on the glutathione system were examined in PDL 20 (early passage) and PDL 50 (late passage) human lung fibroblasts (TIG-1). Methods: To determine cytosolic glutathione peroxidase (GSH-Px) activities, glutathione reductase (GR) activities, oxidized glutathione (GSSG) concentrations, and GSSG/reduced glutathione (GSH) ratios, WSP and hydrocortisone (HC) treatments of TIG-1 cells (PDL 20→50 and PDL 50→75) were performed for 40 days. We also investigated the GSSG concentrations and GR activities in PDL 20 cells that were continuously treated with WSP until PDL 39 and 55. Results: GSSG concentrations decreased in WSP- and HC-treated PDL 50→75 cells. The GSSG/GSH ratios in PDL 50→75 cells became low after the treatments. Increases in GR activities were observed in treated PDL 50→75 cells. The decline in the GSSG concentration of PDL 50→75 cells correlated with the increase in GR activity. The GSSG levels in control cells were higher following cellular age, whereas the levels in treated cells were lower than those in the control. The studies on cellular age-related changes indicated that greater increases in GR activity were found in treated cells than in the control. Conclusion: These results indicated that WSP influences the GSSG concentration that is associated with cellular aging, but the mechanism of GSSG reduction by WSP remains unknown. The enhancement of glutathione status following WSP treatment may be related to the delay in the cellular aging.
Subject(s)
Humans , Glutathione Disulfide , Cell Biology , GlutathioneABSTRACT
Objective: There are several reports of cellular-aging-dependent alterations in the antioxidant capacity of human fibroblasts. Fibroblasts show slower the growth rate at late passages (referred to hereafter as old cells) than at early passages (referred to hereafter as young cells). Antioxidants may control cellular growth by modulating reactive oxygen species (ROS). Methanolic extracts from broad beans (MEBB) contain phenolic compounds and have ROS-scavenging activities. In this study, we investigated the effects of MEBB on cellular growth and antioxidant levels in normal human lung fibroblasts. Methods: To determine cytosolic superoxide dismutase (SOD) activities, cytosolic glutathione peroxidase (GSH-Px) activities, catalase activities, reduced glutathione (GSH) concentrations, and growth rate, MEBB treatments were performed on young and old cells. Results: In young and old cells treated with 120 μg/ml MEBB, the growth rates increased by 28.1 and 15.2%, respectively, compared with controls. The MEBB treatment of young cells caused a 62.5% increase in SOD activity, but the treatment of old cells caused a 39.5% decrease. The catalase activities of the young and old cells treated with MEBB were equal to those of control cells. Young and old cells treated with MEBB were equal to the control cells in terms of GSH-Px activity. The GSH concentrations in the young and old cells treated with 120 μg/ml MEBB increased by 22.1 and 45.9%, respectively. Conclusion: These studies elucidated a new cellular growth mechanism whereby human lung fibroblasts modulate intracellular GSH levels via the action of MEBB.