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灵芝多糖肽对人脐静脉内皮细胞氧化损伤的保护作用
Protective effects of ganoderma lucidum polysaccharides peptide on human umbilical vein endothelial cells injury by reactive oxygen species
【摘要】 目的研究灵芝多糖肽对人脐静脉内皮细胞(HU-VECs)氧化损伤的保护作用。方法原代培养人脐静脉内皮细胞,CD31免疫荧光法鉴定细胞。以叔丁基氢过氧化物(tBOOH)为氧化剂损伤细胞,造成氧化损伤模型,培养液中加入不同浓度灵芝多糖(6.125、12.5、25、50、100mg·mL-1),以CCK-8法检测细胞存活率;Hoechst333258检测细胞氧化损伤引起的凋亡;比色法检测Caspase-3活性变化;电镜检测细胞及细胞器形态学改变。结果灵芝多糖(6.125、12.5、25、50、100mg·mL-1)可减轻叔丁基氢过氧化物对HUVECs的氧化损伤,CCK-8检测灵芝多糖给药组,HUVEC细胞存活率增加。Hoechst33258检测给药组细胞凋亡数量较损伤组减少。比色法检测损伤组Caspase-3活性明显增高,给药组较损伤组减少。电镜可见灵芝多糖给药组减轻细胞器的氧化损伤,减少细胞凋亡。结论灵芝多糖肽(GLPP)对人脐静脉内皮细胞氧化损伤具有保护作用。
【Abstract】 Aim To study the protective effects of Ganoderma lucidum polysaccharides peptide(GLPP)on the human umbilical vein endothelial cells(HUVECs)injured by reactive oxygen species(ROS),derived from tert-butylhydroperoxide(t-BOOH).Methods Vascular endothelial cells were obtained from infant umbilical cord for primary cultivation,identified by endothelial cell adhesion molecule-1(CD31)by fluorescence microscope.HUVECs were injured by ROS,derived from t-BOOH.The survival rate of cells was measured by Cell Counting Kit-8 assay.Hoechst 33258 nucleus staining was used to observe the early apoptosis injury by ROS.And the activation of Caspase-3 was detected by spectrophotometry.Electron microscopy was used to show the morphological changes in HUVECs.Results GLPP(6.125,12.5,25,50,100 mg·L-1)could inhibit the apoptosis of HUVECs by ROS.The survival rate of HUVECs was increased by GLPP and the percentage of cell apoptosis was decreased by it too.GLPP decreased the activation of Caspase-3 of HUVECs up-regulated by ROS.Electron microscope showed that the organelle such as mitochondria injury by ROS could be relieved by GLPP.Conclusion GLPP can prevent human umbilical vein endothelial cells from oxidative injury.
【Key words】 ganoderma lucidum polysaccharides peptide; human umbilical vein endothelial cells; oxidative injury; antioxidants; tertbutylhydroperoxide(t-BOOH); Hoechst33258; Caspase-3;
- 【文献出处】 中国药理学通报 ,Chinese Pharmacological Bulletin , 编辑部邮箱 ,2010年05期
- 【分类号】R285
- 【被引频次】39
- 【下载频次】855