摘要
研究红景天苷(SAL)对同型半胱氨酸(Hcy)诱导体外培养的人脐静脉内皮细胞(HUVECs)氧化应激损伤的保护作用及可能机制.选择3-8代生长状态良好的HUVECs用于实验,采用SAL预孵育HUVECs 2h后,再加入Hcy(1 mmol/L)共孵育12 h诱导内皮细胞氧化应激损伤.采用MTT和LDH法分析细胞损伤,荧光探针法DCFH-DA分析细胞内活性氧(ROS)水平,酶联免疫吸附法分析细胞内丙二醛(MDA),超氧化物歧化酶(SOD)的含量,qRT-PCR技术检测Nrf2、HO-1、NQO1的基因表达.结果显示,Hcy显著引起血管内皮细胞损伤,而10、50μmol/L的红景天苷干预给药后,能够提高Hcy诱导的细胞存活率,明显降低LDH、MDA的水平,增加胞内SOD活性.与Hcy处理组相比,SAL干预给药能够降低Hcy所致细胞内ROS的增高,上调Nrf2、抗氧化酶基因HO-1、醌氧化还原酶NQO1的mRNA表达.本研究表明,红景天苷有效抑制Hcy诱导的HUVECs氧化应激损伤,其作用与红景天苷增强细胞清除自由基能力,调控Nrf2信号有关.
To analyze the protective effect of salidroside against oxidative stress of HUVECs induced by homocysteine and the mechanism involved,HUVECs was selected for the experiment which grew well for 3 to 8 generations,and the model of oxidative stress damage of endothelial cells was established by Hcy.The experiment was divided into normal control group,model group(Hcy,1 mmol/L),salidroside high dose group(SAL,50μmol/L)and salidroside low dose group(SAL,10μmol/L).The cell model of oxidative stress damage was established by incubating with Hcy(1 mmol/L)for 12 h after SAL intervention for 2 h in each group.Cell injury was analyzed by MTT and LDH,intracellular reactive oxygen species(ROS)level was analyzed by fluorescence probe DCFH-DA,malondialdehyde(MDA)and superoxide dismutase(SOD)activity in cells was analyzed by enzyme-linked immunosorbent assay,and the gene expression of Nrf2,HO-1 and NQO1 was detected by RT-PCR.The results showed that Hcy significantly induced vascular endothelial cell injury,but 10 and 50μmol/L salidroside could increase the cell survival rate while the levels of LDH and MDA were significantly decreased and the activity of intracellular SOD was increased.Compared with Hcy treatment group,SAL intervention could reduce the increase of intracellular ROS induced by Hcy and up-regulate the mRNA expression of Nrf2,antioxidant enzyme gene HO-1 and NQO1.These results suggested salidroside has a certain protective effect on HUVECs oxidative stress induced by Hcy,which may be related to its enhancement of free radical scavenging ability and regulation of Nrf2 signal.
作者
张彦燕
何丽
黄梅
姜丰
付凌云
李佳川
ZHANG Yan-yan;HE Li;HUANG Mei;JIANG Feng;FU Ling-yun;LI Jia-chuan(School of Pharmacy,Guizhou Medical University,Guiyang 550025,China;Key Laboratory of Optimal Utilization of Natural Medicine Resource,Guizhou Medical University,Guiyang 550025,China;School of Pharmacy,Southwest Minzu University,Chengdu 610041,China)
出处
《西南民族大学学报（自然科学版）》
CAS
2020年第4期349-353,共5页
Journal of Southwest Minzu University(Natural Science Edition)
基金
国家自然科学基金项目(U1812403-4-4)
贵州省科学技术基金计划(黔科合基础[2016]1128)
中央高校基本科研业务费专项基金项目(2017NZYQN35)。
