摘要
Objective: To investigate the protective effect and underlying mechanism(s) of icariin(ICA) in preventing hydrogen peroxide(H_2O_2)-induced vascular endothelial cell injury via endoplasmic reticulum stress(ERS).Methods: To study the effects of ICA on H_2O_2-induced damage, we used the cell counting kit-8 assay to detect cell viability and the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay to determine cell adhesion and apoptosis, respectively. Spectrophotometry and enzyme-linked immunosorbent assay were used to measure the expression levels of superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px). Subsequently, glucose-regulated protein 78(GRP78), activating transcription factor-4(ATF4) and eukaryotic initiation factor-2 a(eIF2 a) were detected using Western blotting.Results: In human umbilical vein endothelial cells, different concentrations of ICA exhibited multiple effects, including reduced H_2O_2 damage, improved cell viability and adhesion, reduced cell apoptosis and increased SOD and GSH-Px activity. Among the ICA concentrations used, only the H_2O_2+ 100 lmol/L ICA group had significant differences compared to the H_2O_2 group. ERS activators H_2O_2 and DL-dithiothreitol(DTT) significantly increased GRP78, ATF4 and eIF2 a expressions, decreased cell activity and reduced SOD and GSH-Px activity. In contrast, the H_2O_2+ 100 lmol/L ICA and H_2O_2+ 100 lmol/L ICA + DTT groups had significant inhibitory effects on the expressions of GRP78,ATF4 and eIF2 a proteins, showing enhanced cell viability and SOD and GSH-Px activity.Conclusion: The results showed the dose-dependent effects of ICA against H_2O_2-induced injury in vascular endothelial cells. The inhibition of GRP78, ATF4 and eIF2 a protein expressions in the ERS, and the subsequent alleviation of oxidative stress damage, might be the molecular mechanism.
Objective: To investigate the protective effect and underlying mechanism(s) of icariin(ICA) in preventing hydrogen peroxide(H_2O_2)-induced vascular endothelial cell injury via endoplasmic reticulum stress(ERS).Methods: To study the effects of ICA on H_2O_2-induced damage, we used the cell counting kit-8 assay to detect cell viability and the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay to determine cell adhesion and apoptosis, respectively. Spectrophotometry and enzyme-linked immunosorbent assay were used to measure the expression levels of superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px). Subsequently, glucose-regulated protein 78(GRP78), activating transcription factor-4(ATF4) and eukaryotic initiation factor-2 a(eIF2 a) were detected using Western blotting.Results: In human umbilical vein endothelial cells, different concentrations of ICA exhibited multiple effects, including reduced H_2O_2 damage, improved cell viability and adhesion, reduced cell apoptosis and increased SOD and GSH-Px activity. Among the ICA concentrations used, only the H_2O_2+ 100 lmol/L ICA group had significant differences compared to the H_2O_2 group. ERS activators H_2O_2 and DL-dithiothreitol(DTT) significantly increased GRP78, ATF4 and eIF2 a expressions, decreased cell activity and reduced SOD and GSH-Px activity. In contrast, the H_2O_2+ 100 lmol/L ICA and H_2O_2+ 100 lmol/L ICA + DTT groups had significant inhibitory effects on the expressions of GRP78,ATF4 and eIF2 a proteins, showing enhanced cell viability and SOD and GSH-Px activity.Conclusion: The results showed the dose-dependent effects of ICA against H_2O_2-induced injury in vascular endothelial cells. The inhibition of GRP78, ATF4 and eIF2 a protein expressions in the ERS, and the subsequent alleviation of oxidative stress damage, might be the molecular mechanism.
引文
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