Objective To investigate the protective effects of Salvianolic acid B(Sal B)on hydrogen peroxide(H2O2)-induced injury in human umbilical vein endothelial cells(HUVECs).Sal B is considered as one of the most active ant...Objective To investigate the protective effects of Salvianolic acid B(Sal B)on hydrogen peroxide(H2O2)-induced injury in human umbilical vein endothelial cells(HUVECs).Sal B is considered as one of the most active anti-oxidant and the major pharmacological component of the herb,Salvia miltiorrhiza.Its beneficial effects include hepatoprotection,elicitation of endothelium-dependent vasodilation,lowering blood pressure in hypertension,inhibition of HIV-1 replication and suppressing inflammatory cytokine-stimulated endothelial adhesiveness to human monocytic cells by its strong antioxidant activities.Methods Treatment with H2O2 significantly decreased the cell viability and increased the lactate dehydrogenase(LDH)leakage that is an apoptotic feature.Pretreatment with Sal B prevented significantly from H2O2-induced cell apoptosis and other damages in a concentration-dependent manner.The mechanism of Sal B protection was studied with two-dimensional gel electrophoresis(2-DE)coupled to hybrid quadrupole time-of-flight mass spectrometry(Q-TOF)mass spectrometer.Results Data base searching implicated glucose-regulated protein 78(GRP78),a central regulator for ER stress,was up-regulated in Sal B-exposed HUVECs.After exposure to Sal B,the level of activating transcription factor 4(ATF4)was raised,with a transient phosphorylation of the α subunit of eukaryotic translation initiation factor(eIF2α).Knock-down of GRP78 by siRNA significantly reduced protective effects of Sal B.Conclusions These results suggest that Sal B-induced GRP78 up-regulation via phosphorylation of eIF2α and resultant translation of ATF4.And up-regulation of ER chaperones induced by Sal B may play an important role in protecting human endothelial cells from oxidative stress-induced cellular damage.展开更多
文摘Objective To investigate the protective effects of Salvianolic acid B(Sal B)on hydrogen peroxide(H2O2)-induced injury in human umbilical vein endothelial cells(HUVECs).Sal B is considered as one of the most active anti-oxidant and the major pharmacological component of the herb,Salvia miltiorrhiza.Its beneficial effects include hepatoprotection,elicitation of endothelium-dependent vasodilation,lowering blood pressure in hypertension,inhibition of HIV-1 replication and suppressing inflammatory cytokine-stimulated endothelial adhesiveness to human monocytic cells by its strong antioxidant activities.Methods Treatment with H2O2 significantly decreased the cell viability and increased the lactate dehydrogenase(LDH)leakage that is an apoptotic feature.Pretreatment with Sal B prevented significantly from H2O2-induced cell apoptosis and other damages in a concentration-dependent manner.The mechanism of Sal B protection was studied with two-dimensional gel electrophoresis(2-DE)coupled to hybrid quadrupole time-of-flight mass spectrometry(Q-TOF)mass spectrometer.Results Data base searching implicated glucose-regulated protein 78(GRP78),a central regulator for ER stress,was up-regulated in Sal B-exposed HUVECs.After exposure to Sal B,the level of activating transcription factor 4(ATF4)was raised,with a transient phosphorylation of the α subunit of eukaryotic translation initiation factor(eIF2α).Knock-down of GRP78 by siRNA significantly reduced protective effects of Sal B.Conclusions These results suggest that Sal B-induced GRP78 up-regulation via phosphorylation of eIF2α and resultant translation of ATF4.And up-regulation of ER chaperones induced by Sal B may play an important role in protecting human endothelial cells from oxidative stress-induced cellular damage.
文摘目的探究丹酚酸B(Sal B)抑制H_2O_2诱导的细胞衰老作用。方法首先使用不同浓度的Sal B(10-50μM)处理HaCaT细胞,检测细胞的存活率。然后再用浓度为100μM至500μM的H_2O_2孵育18小时检测细胞增殖情况。HaCaT细胞经Sal B处理18小时之后,提取RNA,通过实时荧光定量实验检测SOD1、SOD2、VDR和RARG等抗氧化基因的表达量。结果 Sal B对HaCaT细胞的增殖无抑制作用,但可抑制H_2O_2诱导细胞衰老,且呈剂量依赖性,加入Sal B后H_2O_2(500μM)诱导的活细胞比例由17%提高至58%。Sal B可上调SOD1、SOD2、VDR在HaCaT细胞中的表达。结论 Sal B上调SOD1、SOD2、VDR从而抑制H_2O_2诱导的HaCaT细胞衰老。
