摘要
目的:探讨芍药醇(PAE)对叔丁基过氧化氢(TBHP)损伤的血管内皮细胞的保护作用及其机制。方法:体外培养人脐静脉内皮细胞(HUVECs)分为对照组(Control组)、造模组(TBHP组)和PAE处理组(各浓度PAE+THBP组);用CCK8法及乳酸脱氢酶(LDH)检测各组HUVECs细胞增殖抑制情况,Transwell实验检测细胞的迁移能力,流式细胞仪和Cleaved-caspase 3免疫荧光染色检测HUVECs细胞凋亡情况。结果:与Control组比较,TBHP组细胞存活率和细胞迁移数明显降低,细胞凋亡率、LDH释放量和Cleaved-caspase 3蛋白的表达明显升高(均P<0.05)。与TBHP组比较,PAE处理组细胞的存活率和细胞迁移数均随着给药浓度的增加而逐渐升高,细胞凋亡率、LDH释放量和Cleaved-caspase 3蛋白的表达均随着给药浓度的增加而逐渐下降(均P<0.05)。结论:PAE通过抑制TBHP诱导的HUVECs细胞存活率降低、LDH释放量升高、细胞凋亡增多、Cleaved-caspase 3蛋白表达的上调,发挥其对TBHP损伤的血管内皮细胞的保护作用,且呈剂量依赖性。
Objective:To investigate the protective effects and mechanism of paeonol(PAE)on vascular endothelial cells injured by tert-butyl hydroperoxide(TBHP).Methods:Human umbilical vein endothelial cells(HUVECs)were randomly divided into control group(only treated with 1640 medium),injured group(stimulated by TBHP),and PAE treatment group(pretreated with various dose of PAE followed by TBHP stimulation).CCK-8 method and lactate dehydrogenase(LDH)release assay were used to detect the proliferation of HUVECs in each group.Transwell assay was used to detect the migration ability of HUVECs.Flow cytometry and immunofluorescence staining of Cleaved-caspase 3 were used to detect the apoptosis rate of HUVECs in each group.Results:Compared with the control group,the cell survival rate and cell migration number of the model group were significantly decreased,and the apoptosis rate,LDH release rate and the expression of Cleaved-caspase 3 protein were significantly increased(P<0.05).Compared with injured group,the survival rate and cell migration number of the PAE treatment group gradually increased in a dose-dependent manner;moreover,the apoptosis rate,LDH release rate and the expression of Cleaved-caspase 3 protein decreased in a dose-dependent manner(P<0.05).Conclusion:Paeonol can inhibit the TBHP-induced HUVECs viability due to the increase in LDH release and apoptosis,up-regulate the expression of Cleaved-caspase 3,and exert its protective effect on vascular endothelial cells injured by TBHP in a dose-dependent manner.
作者
潘达
胡飞虹
金灿
闻浩
孟伟阳
朱烈烈
陈大庆
PAN Da;HU Feihong;JIN Can;WEN Hao;MENG Weiyang;ZHU Lielie;CHEN Daqing(Department of Emergency Medicine,the Second Affiliated Hospital&Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou 325027,China;Department of Spine Surgery,the Second Affiliated Hospital&Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou 325027,China.)
出处
《温州医科大学学报》
CAS
2019年第11期797-801,806,共6页
Journal of Wenzhou Medical University
基金
浙江省自然科学基金资助项目(LY19H150001)