Department of Gerontology, the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310002, China.
Department of Cardiology, the Affiliated Hangzhou Hospital, Nanjing Medical University, Hangzhou 310006, China.
Exp Gerontol. 2020 Oct 1;139:111053. doi: 10.1016/j.exger.2020.111053. Epub 2020 Aug 5.
Caloric restriction (CR) is an important means to delay senescence, and glucose restriction is one of the measures to achieve CR. On the basis of our previous work and bioinformatics analysis, we hypothesized that glucose restriction can up-regulate autophagy, inhibit senescence and promote proliferation via the AMPK/SIRT1-FOXA3-Beclin1 pathway in human umbilical vein endothelial cells (HUVECs). We found that compared with 5.5 mmol/L and 25 mmol/L glucose, 2.5 mmol/L glucose restriction significantly reduced senescence, enhanced autophagy, increased migration speed, relieved G/G phase arrest and enhanced proliferation of HUVECs. Furthermore, glucose restriction up-regulated AMPKα1, SIRT1, FOXA3 and Beclin1 expression in HUVECs. Additionally, we demonstrated that AMPKα1 phosphorylated FOXA3 at S170 and S305 in the cytoplasm and promoted FOXA3 nuclear translocation under glucose restriction. FOXA3 in the nucleus was deacetylated by SIRT1 at K214 and K221. Deacetylated FOXA3 specifically bound to +109 C in the Beclin1 transcriptional regulatory region, and significantly enhanced Beclin1 transcription and expression. siRNA knock down of AMPKα1, SIRT1, FOXA3 or Beclin1 expression impaired the glucose restriction-induced inhibition of senescence, enhanced autophagy, increased migration, and induced proliferation of HUVECs. This study confirmed that glucose restriction can enhance autophagy, inhibit senescence, and enhance proliferation of HUVECs through the AMPK/SIRT1-FOXA3-Beclin1 pathway.
热量限制（CR）是延缓衰老的重要手段，而葡萄糖限制是实现 CR 的措施之一。基于我们之前的工作和生物信息学分析，我们假设葡萄糖限制可以通过 AMPK/SIRT1-FOXA3-Beclin1 通路上调自噬、抑制衰老并促进人脐静脉内皮细胞（HUVEC）的增殖。我们发现，与 5.5mmol/L 和 25mmol/L 葡萄糖相比，2.5mmol/L 葡萄糖限制显著减少了衰老，增强了自噬，增加了迁移速度，缓解了 G/G 期停滞，并增强了 HUVEC 的增殖。此外，葡萄糖限制上调了 HUVEC 中 AMPKα1、SIRT1、FOXA3 和 Beclin1 的表达。此外，我们证明了 AMPKα1 在葡萄糖限制下在细胞质中磷酸化 FOXA3 的 S170 和 S305 并促进 FOXA3 核易位。SIRT1 在 K214 和 K221 处使核内 FOXA3 去乙酰化。去乙酰化的 FOXA3 特异性结合 Beclin1 转录调控区的+109 C，显著增强 Beclin1 转录和表达。siRNA 敲低 AMPKα1、SIRT1、FOXA3 或 Beclin1 表达会损害葡萄糖限制诱导的衰老抑制、增强自噬、增加迁移和诱导 HUVEC 增殖。这项研究证实，葡萄糖限制可以通过 AMPK/SIRT1-FOXA3-Beclin1 通路增强 HUVEC 的自噬、抑制衰老和促进增殖。
