Guo Chao, Yin Ying, Ma Zhongying, Xu Fangqin, Wang Shiquan
Xi'an People's Hospital (Xi'an Fourth Hospital), Xi'an, China.
Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Xi'an, China.
Heliyon. 2024 Sep 3;10(17):e37276. doi: 10.1016/j.heliyon.2024.e37276. eCollection 2024 Sep 15.
The present study aims to evaluate the impact of Astilbin (AST) on cortical neuron survival under conditions of oxygen-glucose deprivation and reoxygenation (OGD/R) and determine the role of NF-E2-related factor 2 (Nrf2) in this process.
Primary neurons were pre-treated with various concentrations of AST for 8 h before OGD induction. Cell viability and lactate dehydrogenase (LDH) leakage were assessed to determine the optimal concentration. Biomarkers related to oxidative stress, antioxidant enzyme activities, and apoptosis were evaluated at 24 h post-OGD/R. To investigate the involvement of Nrf2 in AST-mediated neuroprotection, we conducted molecular docking and microscale thermophoresis analyses, as well as examined the expression levels of Nrf2 and its regulatory genes including heme oxygenase-1(HO-1), (NAD(P)H: quinone oxidoreductase 1 (NQO-1), and peroxiredoxin 1 (Prdx1). Additionally, lentivirus-mediated knockdown of Nrf2 and overexpression of Nrf2 with L-sulforaphane (SFN) were performed, followed by an assessment of cell viability, oxidative stress, antioxidant enzyme activities and apoptosis.
Pre-treatment with AST reduced oxidative stress levels while increasing antioxidant enzyme activities and mitigating neuronal apoptosis. After OGD/R exposure, AST upregulated nuclear Nrf2 expression and increased the expression of HO-1, NQO-1 and Prdx1 in the cytoplasm. However, the knockdown of Nrf2 abolished the antioxidative and protective effects exerted by AST treatment. Conversely, combining AST with the Nrf2 agonist SFN demonstrated an enhancement in the protective effects provided by AST. These results demonstrate that Nrf2-dependent antioxidant responses contribute to AST-induced tolerance against neuronal injury caused by OGD/R injury.
Overall findings support the ability of AST to protect primary neurons from OGD/R-induced damage through activation of Nrf2-dependent antioxidant responses.
本研究旨在评估落新妇苷（AST）在氧糖剥夺及复氧（OGD/R）条件下对皮质神经元存活的影响，并确定核因子E2相关因子2（Nrf2）在此过程中的作用。
原代神经元在诱导OGD前用不同浓度的AST预处理8小时。评估细胞活力和乳酸脱氢酶（LDH）泄漏情况以确定最佳浓度。在OGD/R后24小时评估与氧化应激、抗氧化酶活性和细胞凋亡相关的生物标志物。为研究Nrf2在AST介导的神经保护中的作用，我们进行了分子对接和微量热泳分析，并检测了Nrf2及其调控基因包括血红素加氧酶-1（HO-1）、烟酰胺腺嘌呤二核苷酸磷酸醌氧化还原酶1（NQO-1）和过氧化物还原酶1（Prdx1）的表达水平。此外，进行慢病毒介导的Nrf2敲低和用莱菔硫烷（SFN）过表达Nrf2，随后评估细胞活力、氧化应激、抗氧化酶活性和细胞凋亡。
AST预处理降低了氧化应激水平，同时增加了抗氧化酶活性并减轻了神经元凋亡。在OGD/R暴露后，AST上调了核Nrf2表达，并增加了细胞质中HO-1、NQO-1和Prdx1的表达。然而，Nrf2的敲低消除了AST处理所发挥的抗氧化和保护作用。相反，将AST与Nrf2激动剂SFN联合使用显示AST提供的保护作用增强。这些结果表明，Nrf2依赖性抗氧化反应有助于AST诱导的对OGD/R损伤所致神经元损伤的耐受性。
总体研究结果支持AST通过激活Nrf2依赖性抗氧化反应保护原代神经元免受OGD/R诱导损伤的能力。
