冯 笑，杨 阳，熊红燕，范崇熙，狄守印，胡 伟，段维勋，梁洪亮，金振晓，俞世强



SIRT1信号通路介导紫檀芪抗内皮细胞抗缺血再灌注损伤的机制研究

冯 笑，杨 阳，熊红燕，范崇熙，狄守印，胡 伟，段维勋，梁洪亮，金振晓，俞世强

［摘要］：目的 研究不同浓度紫檀芪(PTE)对人脐静脉内皮细胞(HUVECs)缺血再灌注损伤的保护作用及其可能的机制。方法 HUVECs经1 μM/L和2 μM/L的PTE处理后,接受模拟缺氧复氧(SIR)损伤(缺氧70 min,复氧4 h),采用CCK-8法检测细胞活力,使用酶活性测定法检测细胞培养液中丙二醛(MDA)的释放量和超氧化物歧化酶(SOD)的水平,使用western blot法检测SIRT1、Ac-FOXO1、Bax和Bcl-2的表达情况,使用SIRT1阻断剂EX527观察SIRT信号通路在这一过程中发挥的作用。结果 SIR处理可以抑制HUVECs活性,抑制SIRT1表达,上调Ac-FOXO1表达,下调Bcl-2/Bax比例(与Control组比较,P＜0.05)。细胞培养基中MDA含量上升,SOD水平下降(与Control组比较,P＜0.05)。1 μM/L和2 μM/L的PTE处理均可以发挥细胞保护作用,提高细胞活力,降低了MDA水平,提高了SOD水平(与SIR组比较,P＜0.05)。此外,PTE处理上调了SIRT1表达,抑制了Ac-FOXO1表达,并且提高Bcl-2/Bax比例(与SIR组比较,P＜0.05)。EX527处理后细胞活力下降,凋亡增加(与SIR＋PTE组比较,P＜0.05)。结论 PTE可以保护HUVECs抵抗缺血再灌注损伤,其机制是激活SIRT1/FOXO1保护性信号通路,抑制HUVECs凋亡。

［关键词］：紫檀芪；人脐静脉内皮细胞；缺血再灌注损伤；沉默信息调控因子1

Pterostilbene alleviates HUVECs ischemia reperfusion injury via activating SIRT1 and suppressing cell apoptosis

Feng Xiao,Yang-Yang,Xiong Hong-yan,Fan Chong-xi,Di Shou-yin,Hu Wei,Duan Wei-xun,Liang Hong-liang,Jin Zhen-xiao,Yu Shi-qiang
Department of Cardiovascular Surgery,Xijing Hospital,Fourth Military Medical University,Shaan＇xi Xi＇an 710032,China
Corresponding author:Jin Zhen-xiao,Email:13571921011@ 163.com

［Abstract］：Objective To explore the effect of pterostilbene on simulated ischemia reperfusion(SIR) injury of HUVECs and the roles of SIRT1 in this process.Methods The cultured HUVECs,pretreated with PTE(1 or 2 μM/L) for 2h,were subjected to SIR.Then cell counting kit-8 was used to detect cell viability,and specialized kits were used to detect the concentration of malondialdehyde(MDA) and superoxide dismutase(SOD) in the cell culture medium.The expression of Silent information regulator 1(SIRT1),Acetylated fork head transcription factor 1(Ac-FOXO1),Bcl-2 and Bax were quantified using western blotting.The specific inhibitor of SIRT1 EX527 was used to investigate the roles of SIRT1 in this process.Results SIR treatment reduced cell viability significantly(from 1.219±0.021 to 0.593±0.036),down-regulated SIRT1 expression,up-regulated Ac-FOXO1 expression and reduced the Bcl-2/Bax ratio(vs the control group,P＜0.05).Both 1μM/Land 2 μM/L PTE increased the cell viability(to 0.762±0.029和0.860±0.019),down-regulated the MDA level(to 14.16±1.62 and 11.23±1.35) and up-regulated the SOD level(to 64.16±3.97 and 86.06±5.48 )(vs the SIR group,P＜0.05).Western blot analysis showed that SIRT1 expression increased and Ac-FOXO1 expression decreased.The Bcl-2/Bax ratio was increased(vs the SIR group,P＜0.05).EX527 treatment reduced the cell viability(from 0.743±0.031 to 0.6140±0.025)(vs the SIR＋PTE group,P＜0.05).Conclusion PTE treatment may activate SIR injury through activating SIRT1-Ac-FOXO1 pathway and suppressing cell apoptosis.

［Key words］： Pterostilbene；HUVECs；Ischemia reperfusion injury；Silent information regulation 1

心血管疾病是当今社会危害人类生命健康的首要杀手,心肌缺血再灌注损伤是重要的损伤类型,内皮细胞损伤在这个病理过程中发挥着重要的作用[1-3]。内皮细胞构成了血管和周围组织的屏障,心肌缺血再灌注损伤可以促使内皮细胞发生凋亡,最终损伤向心肌重构和心衰的方向发展[3-4]。……