落新妇苷缓解过氧化氢诱导的氧化应激和细胞凋亡减轻内皮细胞损伤  

Astilbin alleviates hydrogen peroxide-induced oxidative stress and cell apoptosis to reduce endothelial cell injury

作　　者：马芬芬 徐海涛 王语嫣 王彦 MA Fenfen;XU Haitao;WANG Yuyan;WANG Yan(Department of Cardiac Function Testing,the Second People′s Hospital of Lianyungang Affiliated to Kangda College of Nanjing Medical University,Lianyungang,Jiangsu 222000,China;Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening,School of Pharmacy,Jiangsu Ocean University,Lianyungang,Jiangsu 222000;Department of Clinical Laboratory,the Second People′s Hospital of Lianyungang Affiliated to Kangda College of Nanjing Medical University)

机构地区：[1]南京医科大学康达学院附属连云港市第二人民医院心脏功能检查科,江苏连云港222000 [2]江苏海洋大学药学院·江苏海洋生物资源与环境重点实验室,江苏连云港222000 [3]南京医科大学康达学院附属连云港市第二人民医院检验科

出　　处：《徐州医科大学学报》2025年第3期179-185,共7页Journal of Xuzhou Medical University

基　　金：江苏省卫健委医学科研指导性项目(Z2022070);南京医科大学康达学院科研发展基金(KD2023KYJJ062)。

摘　　要：目的探讨落新妇苷对过氧化氢(H_(2)O_(2))诱导的人脐静脉内皮细胞(HUVEC)损伤的保护作用及机制。方法采用CCK8法筛选落新妇苷实验浓度。将HUVEC分为对照组、H_(2)O_(2)组和中/高浓度(50、100μmol/L)落新妇苷组。以H_(2)O_(2)(400μmol/L)诱导建立HUVEC氧化损伤模型,检测细胞活力、丙二醛(MDA)含量、谷胱甘肽过氧化物酶(GSH-Px)、总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、活性细胞内活性氧(ROS)、一氧化氮(NO)的水平;实时荧光定量PCR(RT-qPCR)检测细胞中fas、caspase-8、Bax、Bcl-2、caspase-3的mRNA表达水平;Western blot检测细胞中Bax、Bcl-2、Cyto-C、p53、cleaved-caspase-3的蛋白表达水平。结果CCK8结果显示,落新妇苷在1~100μmol/L浓度范围内,对HUVEC无毒性。与对照组相比,H_(2)O_(2)组细胞存活率显著降低,细胞皱缩,MDA含量升高,GSH-Px、T-AOC、SOD活性减弱,ROS累积、NO释放增加,Cyto-C、Bax、p53、caspase-3转录和蛋白表达水平升高,而Bcl-2转录和蛋白表达呈下降趋势。与H_(2)O_(2)组相比,落新妇苷组HUVEC形态恢复,MDA含量减少,GSH-Px、T-AOC和SOD活性增强,ROS、NO释放减少,Bcl-2蛋白表达呈升高趋势,但Cyto-C、Bax、p53、caspase-3表达水平被抑制。结论落新妇苷对H_(2)O_(2)所致内皮细胞损伤具有明显的保护作用,其机制可能与改善细胞内氧化应激水平和凋亡相关。Objective To investigate the protective effect of astilbin on hydrogen peroxide(H_(2)O_(2))-induced injury in human umbilical vein endothelial cells(HUVEC)and related mechnisms.Methods The experimental concentration of astilbin was screened by CCK8 assay.Endothelial cells were divided into four groups:control group,H_(2)O_(2) group,and medium/high-concentration(50 and 100μmol/L)astilbin groups.An oxidative injury model of endothelial cells was established by H_(2)O_(2) induction at 400μmol/L.Cell viability,malondialdehyde(MDA)content,activities of glutathione peroxidase(GSH-Px),total antioxidant capacity(T-AOC)and superoxide dismutase(SOD),intracellular reactive oxygen species(ROS),and nitric oxide(NO)were measured.The mRNA expression of fas,caspase-8,Bax,Bcl-2,and caspase-3 was detected by real-time fluorescence quantitative PCR(RT-qPCR).The protein expression of Bax,Bcl-2,Cyto-C,p53,and cleaved-caspase-3 was measured by Western blot.Results CCK8 results showed that astilbin exerted no toxicity to HUVEC within the concentration range of 1-100μmol/L.Compared with the control group,the H_(2)O_(2) group showed significantly reduced cell viability,with cell shrinkage,and presented increases in MDA content,decreases in GSH-Px,T-AOC and SOD activities,and increases in ROS accumulation and NO release.The H_(2)O_(2) group also exhibited increased mRNA and protein levels of Cyto-C,Bax,p53,and caspase-3,and decreased mRNA and protein levels of Bcl-2.Compared with the H_(2)O_(2) group,the astilbin group showed improved cell morphology,reduced MDA content,enhanced GSH-Px,T-AOC,and SOD activities,reduced ROS and NO release,and an increasing trend in Bcl-2 protein expression,and inhibited expression of Cyto-C,Bax,p53,and caspase-3.Conclusions Astilbin exert significant protective effect on H_(2)O_(2)-induced endothelial cell injury,which may be related to improved intracellular oxidative stress and apoptosis.

关 键 词：落新妇苷 内皮细胞 氧化应激 细胞凋亡 活性氧 过氧化氢 

分 类 号：R543[医药卫生—心血管疾病]