Translational Medicine Research Center, Shanxi Medical University, Taiyuan, Shanxi, PR China.
National Key Disciplines, Key Laboratory for Cellular Physiology of Ministry of Education, Shanxi Medical University, Taiyuan, Shanxi, PR China.
J Biochem Mol Toxicol. 2022 Jan;36(1):e22939. doi: 10.1002/jbt.22939. Epub 2021 Oct 31.
The anti-senescence function of genistein is related to inhibiting oxidative stress, however, the mechanism has not been clarified. The present study aimed to explore the effects of genistein on oxidized low-density lipoprotein (ox-LDL)-induced endothelial senescence and the role of the sirtuin-1 (SIRT1)-66-kDa Src homology 2 domain-containing protein (p66Shc)-forkhead box protein O3 (Foxo3a) pathways in the process. In this paper, human umbilical vein endothelial cells were pretreated with 1000 nM genistein for 30 min and then incubated with 50 mg/L ox-LDL for another 12 h; meanwhile, the functions of adenovirus-mediated overexpression of p66shc and small interfering RNA-mediated silencing of SIRT1 were investigated. Results showed that genistein pretreatment alleviated ox-LDL-induced mitochondrial reactive oxygen species, the levels of oxidatively modified DNA (8-OHdG) and pai-1, and the activity of SA-β-gal, which was associated with mitigating p66shc. Further studies indicated the inhibitory effect of genistein on p66shc was correlated with suppressing the acetylation and phosphorylation of p66shc, and ameliorating its mitochondrial translocation by activating SIRT1. Moreover, the inactivated p66shc could enhance the activity of Foxo3a via restraining the phosphorylation and triggering nucleus accumulation. The study demonstrates genistein could prevent ox-LDL-induced mitochondrial oxidative stress and senescence through the SIRT1-p66shc-Foxo3a pathways.
染料木黄酮的抗衰老功能与抑制氧化应激有关，但具体机制尚未阐明。本研究旨在探讨染料木黄酮对氧化型低密度脂蛋白（ox-LDL）诱导的内皮细胞衰老的影响，以及在这一过程中，沉默信息调节因子 1（SIRT1）-66 kDaSrc 同源 2 结构域蛋白（p66Shc）-叉头框蛋白 O3（Foxo3a）通路的作用。本文中，人脐静脉内皮细胞先用 1000 nM 染料木黄酮预处理 30 min，再用 50 mg/L ox-LDL 孵育 12 h；同时，还研究了过表达 p66Shc 的腺病毒和沉默 SIRT1 的小干扰 RNA 的功能。结果表明，染料木黄酮预处理可减轻 ox-LDL 诱导的线粒体活性氧、氧化修饰的 DNA（8-OHdG）和 pai-1 的水平，以及 SA-β-半乳糖苷酶的活性，这与减轻 p66Shc 有关。进一步的研究表明，染料木黄酮对 p66Shc 的抑制作用与抑制 p66Shc 的乙酰化和磷酸化以及通过激活 SIRT1 改善其线粒体易位有关。此外，失活的 p66Shc 可以通过抑制磷酸化和触发核内积累来增强 Foxo3a 的活性。本研究表明，染料木黄酮可以通过 SIRT1-p66Shc-Foxo3a 通路预防 ox-LDL 诱导的线粒体氧化应激和衰老。
