College of Pharmacy, Henan University of Chinese Medicine, Zhengzhou, 450046, China; The Engineering and Technology Center for Chinese Medicine Development of Henan Province, Zhengzhou, 450046, China.
College of Pharmacy, Henan University of Chinese Medicine, Zhengzhou, 450046, China.
Arch Biochem Biophys. 2022 Oct 30;729:109377. doi: 10.1016/j.abb.2022.109377. Epub 2022 Aug 20.
Alzheimer's disease (AD) is a progressive disease with a long duration and complicated pathogenesis. Thymidine (Thy) and 2'-deoxyuridine (2'-De) are pyrimidines nucleotides that are associated with nervous system diseases. However, it remains unclear whether Thy and 2'-De exert neuroprotective effects in AD. Therefore, this study was conducted to explore the interventional effects and mechanisms of Thy and 2'-De on the Aβ-induced brain injury. Donepezil (Do, 10 mg/kg/d), Thy (20 mg/kg/d), and 2'-De (20 mg/kg/d) were administered for 4 weeks after the injection of Aβ peptides (200 μM, i.c.v.) to mice. UPLC-MS/MS method was performed to quantify Thy and 2'-De in the hippocampus of mice brain. The cognition ability, neuronal and mitochondria damage, and levels of Aβ/Aβ, p-Tau, Na K-ATPase, apoptosis, oxidative stress, immune cells, and Iba 1 were measured in Aβ-induced mice. The oxygen consumption (OCR) and extracellular acidification rate (ECAR) were measured using a seahorse analyzer in Aβ-induced N9 cells. Moreover, 2-Deoxy-D-glucose (2-DG), a glycolysis inhibitor, was added to explore the mechanisms underlying the effects of Thy and 2'-De on Aβ-induced N9 cells. The expression of Iba 1 and levels of CD11b and reactive oxygen species (ROS) were measured after treatment with Thy (5 μM) and 2'-De (10 μM) against 2-DG (5 mM) in Aβ-induced N9 cells. The results suggested that Do, Thy, and 2'-De improved the cognition ability, attenuated the damage to hippocampus and mitochondria, downregulated the levels of Aβ/Aβ, p-Tau, Na K-ATPase, apoptosis, oxidative stress, and Iba 1, and regulated the immune response induced by Aβ against the brain injury. Furthermore, Do, Thy, and 2'-De increased ATP production and inhibited glycolysis in Aβ-induced N9 cells. Moreover, 2-DG enhanced the effects of drugs, reduced microglial activation, and attenuated oxidative stress to interfere with Aβ-induced N9 cells. In conclusion, Thy and 2'-De reduced microglial activation and improved oxidative stress damage by modulating glycolytic metabolism on the Aβ-induced brain injury.
阿尔茨海默病（AD）是一种具有长期病程和复杂发病机制的进行性疾病。胸苷（Thy）和 2'-脱氧尿苷（2'-De）是与神经系统疾病相关的嘧啶核苷酸。然而，目前尚不清楚 Thy 和 2'-De 是否对 AD 具有神经保护作用。因此，本研究旨在探讨 Thy 和 2'-De 对 Aβ 诱导的脑损伤的干预作用及其机制。在向小鼠脑内注射 Aβ 肽（200μM，脑室内）后，给予多奈哌齐（Do，10mg/kg/d）、Thy（20mg/kg/d）和 2'-De（20mg/kg/d）4 周。采用 UPLC-MS/MS 法检测小鼠脑内 Thy 和 2'-De 的含量。测量 Aβ 诱导的小鼠的认知能力、神经元和线粒体损伤以及 Aβ/Aβ、p-Tau、Na K-ATPase、凋亡、氧化应激、免疫细胞和 Iba 1 的水平。使用 Seahorse 分析仪测量 Aβ 诱导的 N9 细胞的耗氧量（OCR）和细胞外酸化率（ECAR）。此外，加入 2-脱氧-D-葡萄糖（2-DG），一种糖酵解抑制剂，以探讨 Thy 和 2'-De 对 Aβ 诱导的 N9 细胞的作用机制。在 Aβ 诱导的 N9 细胞中，用 Thy（5μM）和 2'-De（10μM）处理后，测量 Iba 1 的表达和 CD11b 和活性氧（ROS）的水平，然后用 2-DG（5mM）处理。结果表明，Do、Thy 和 2'-De 改善了认知能力，减轻了海马和线粒体的损伤，下调了 Aβ/Aβ、p-Tau、Na K-ATPase、凋亡、氧化应激和 Iba 1 的水平，并调节了 Aβ 诱导的对脑损伤的免疫反应。此外，Do、Thy 和 2'-De 增加了 Aβ 诱导的 N9 细胞中的 ATP 产生并抑制了糖酵解。此外，2-DG 增强了药物的作用，减少了小胶质细胞的激活，并减轻了氧化应激，从而干扰了 Aβ 诱导的 N9 细胞。总之，Thy 和 2'-De 通过调节糖酵解代谢减轻了 Aβ 诱导的脑损伤中小胶质细胞的激活和改善了氧化应激损伤。
