摘要
【目的】本研究拟探讨人参皂苷Rb1通过调节沉默信息调节因子3/超氧化物歧化酶2(Sirt3/SOD2)通路延缓高糖诱导的人脐静脉内皮细胞(HUVEC)衰老的作用和机制。【方法】建立高糖(40 mmol/L葡萄糖)诱导的HUVEC早熟性衰老模型,根据衰老相关β-半乳糖苷酶(SA-β-Gal)阳性率及Ⅰ型纤溶酶原激活物抑制因子(PAI-1)和P16表达评估HUVEC衰老,并采用Annexin V-FITC/PI排除细胞凋亡的发生。采用Western blot检测各组Sirt3与SOD2表达的变化。同时检测细胞内丙二醛(MDA)及SOD2活性的水平。【结果】40 mmol/L葡萄糖处理24 h可成功诱导HUVEC衰老,并未发生细胞凋亡,早熟性衰老的HUVEC SA-β-Gal阳性率明显增多,PAI-1及P16表达增多,Sirt3及SOD2表达均减少,MDA含量升高,SOD2活性降低(P <0.05)。与高糖处理组比较,40μmol/L人参皂苷Rb1预处理延缓HUVEC早熟性衰老,SA-β-Gal阳性率明显减少,PAI-1及P16表达减少,Sirt3及SOD2表达均增多,MDA含量降低,SOD2活性升高(P <0.05)。与人参皂苷Rb1处理组相比,Sirt3特异性抑制剂3-TYP处理后,人参皂苷Rb1保护作用消失,HUVEC PAI-1及P16表达升高,Sirt3及SOD2表达均减少。【结论】人参皂苷Rb1可通过激活Sirt3/SOD2信号通路延缓高糖诱导的HUVEC早熟性衰老。
【Objective】To investigate the effect and mechanism of ginsenoside Rb1 attenuating human umbilical vein endothelial cells(HUVEC) senescence induced by high glucose through Sirt3/SOD2 pathway.【Methods】The senescence of HUVEC induced by high glucose(40 mmol/L)was assessed by senescence-associated β-galactosidase(SA-β-Gal)staining,and the expression of plasminogen activator inhibitor 1(PAI-1)and P16. Annexin V-FITC/PI was performed to measure apoptotic effect. The expression of sirtuins 3(Sirt3)and superoxide dismutase 2(SOD2)was detected by western blot. Meanwhile,the level of intracellular malondialdehyde(MDA)and the activity of SOD2 were measured.【Results】Treatment of HUVEC with high glucose for 24 hours induced premature senescence instead of apoptosis,as indicated by a larger proportion of the cells stained with SA-β-Gal and the up-regulated expression of PAI-1 and P16. Pretreatment of HUVEC with ginsenoside Rb1(40 μmol/L)could reverse endothelial cell senescence,as indicated by the reduced SA-β-Gal positive cells and the down-regulated expression of PAI-1 and P16. Furthermore,ginsenoside Rb1 pretreatment upregulated the protein expression of Sirt3 and SOD2,and eventually increased the activity of SOD2 and decreased the level of MDA.【Conclusion】Ginsenoside Rb1 could antagonize high glucose-induced premature senescence of HUVEC via Sirt3/SOD2 signaling pathway.
作者
柯世业
石光耀
刘定辉
吴琳
温仁辉
朱洁明
钱孝贤
KE Shi-ye;SHI Guang-yao;LIU Ding-hui;WU Lin;WEN Ren-hui;ZHU Jie-ming;QIAN Xiao-xian(Department of Cardiology,The Third Affiliated Hospital,Sun Yat-sen University,Guangzhou 510630,China;Institute of Integrated Traditional Chinese and WesternMedicine,Sun Yat-sen University,Guangzhou 510630,China)
出处
《中山大学学报（医学版）》
CAS
CSCD
北大核心
2019年第3期329-336,共8页
Journal of Sun Yat-Sen University:Medical Sciences
基金
国家自然科学基金(81370447)
广东省科技计划项目(2016A050502014)
