人参皂苷Rb1通过抑制NF-κB p65介导的炎症和氧化应激改善内皮细胞复制性衰老  

Ginsenoside Rb1 Ameliorates the Replicative Senescence of Endothelial Cells by Suppressing Inflammation and Oxidative Stress Via NF-κB p65

作　　者：周彬[1] 吴琳[1] 凌叶盛 余舒杰[1] 刘定辉[1] 柯世业 刘勇[1] 郝宝顺[1] 钱孝贤[1,2] ZHOU Bin;WU Lin;LING Ye-sheng;YU Shu-jie;LIU Ding-hui;KE Shi-ye;LIU Yong;HAO Bao-shun;QIAN Xiao-xian(Department of Cardiology,The Third Affiliated Hospital,Guangzhou 510630,China;Institute of Integrated Traditional Chinese and Western Medicine,Sun Yat-sen University,Guangzhou 510630,China)

机构地区：[1]中山大学附属第三医院心血管内科,广东广州510630 [2]中山大学中西医结合研究所,广东广州510630

出　　处：《中山大学学报（医学版）》2018年第6期835-843,共9页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81370447);广东省医学科研基金(A2017014);广东省科技计划项目(2016A050502014)

摘　　要：【目的】本研究旨在探讨人参皂苷Rb1对内皮细胞复制性衰老的作用和机制。【方法】建立人原代脐静脉内皮细胞(HUVEC)复制性衰老模型,根据细胞形态的变化、衰老相关β-半乳糖苷酶(SA-β-Gal)染色阳性率和纤溶酶原激活物抑制剂1(PAI-1)的表达水平评估HUVEC衰老;将复制性衰老模型细胞分为0、20、40、80、100μmol/L Rb1组,不同浓度人参皂苷Rb1处理衰老细胞48 h后观察衰老指标SA-β-Gal染色和PAI-1蛋白变化,每组3个复孔;采用Western blot方法检测对照组(CPDL 2的HUVEC)、模型组(衰老细胞组)及人参皂苷Rb1组核因子κB p65(NF-κB p65)的活性,每组3个复孔;检测对照组、模型组及人参皂苷Rb1组细胞培养液中氧化应激和炎症指标,每组3个复孔。【结果】累计细胞群体倍增值(CPDL)16的HUVEC可作为复制性衰老模型;与0μmol/L Rb1组相比,80μmol/L Rb1组SA-β-Gal染色阳性细胞数和PAI-1蛋白表达均明显降低(P <0.001);复制性衰老细胞中超氧化物歧化酶(SOD)的活力明显低于对照组(P <0.001),而丙二醛(MDA)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)的含量及NF-κB p65的活性均明显高于对照组(P <0.001),80μmol/L人参皂苷Rb1处理后SOD活力明显增加(P <0.05),MDA、IL-6、TNF-α的产量明显减少(P <0.05),NF-κB p65活性降低(P <0.001)。【结论】人参皂苷Rb1可通过抑制NF-κB p65介导的氧化应激和炎症反应延缓复制性内皮细胞衰老。【Objective】To explore the effect and mechanism of ginsenoside Rb1 on replicative senescence of endothelial cells.【Methods】Establish the replicative senescence model of primary human umbilical vein endothelial cells(HU-VEC):the morphological change of the cells,the proportion of senescence-associatedβ-galactosidase(SA-β-Gal)positive cells and the plasminogen activator inhibitor 1(PAI-1)expression were detected to assess the senescence model;replicative senescence model cells were divided into 0,20,40,80 and 100μmol/L Rb1 group,SA-β-Gal staining positive cells and PAI-1 protein level were measured after treatment of senescent cells with different concentrations of Ginsenoside Rb1 for 48 h,3 multiple pores in each group.NF-κB p65 and p-NF-κB p65 protein expression in normal cell group(CPDL 2 HUVEC),model group(senescent cell group)and ginsenoside Rb1 group were detected by RT-PCR,3 multiple pores in each group;the markers of oxidative stress and inflammation in cell culture solution were measured,3 multiple pores in each group.【Results】HUVEC with CPDL 16 have been chosen as the replicative senescence model in this research.Compared with the 0μmol/L Rb1 group,the SA-β-Gal positive cells and protein expression level of PAI-1 were significantly decreased(P<0.001)in 80μmol/L Rb1 group.Compared with the control group,the production of MDA,IL-6 and TNF-αand NF-κB p65 activity were significantly increased(P<0.001)in model group,but the activity of SOD was significantly decreased(P<0.001).However,Compared with the model group,the activity of SOD was increased(P<0.05),the production of MDA,IL-6 and TNF-αwere decreased(P<0.05)and the activity of NF-κB p65 was decreased(P<0.001)in 80μmol/L Rb1 group.【Conclusions】Ginsenoside Rb1 ameliorates the replicative senescence of endothelial cells by suppressing inflammation and oxidative stress via NF-κB p65.

关 键 词：人参皂苷RB1 炎症 氧化应激 NF-ΚBP65 复制性细胞衰老 

分 类 号：R339.38[医药卫生—人体生理学]