Zhang Pengpeng, Zhang Jie, Ma Chuan, Ma Huiping, Jing Linlin
Department of Pharmacy, the First Affiliated Hospital of Xi'an Jiaotong University, NO.277 Yanta West Road, Yanta District, Xi'an, 710061, Shaanxi, People's Republic of China.
Department of Pharmacy, the 940th Hospital of Joint Logistics Support force of PLA, NO.333 Binhe South Road, Qilihe District, Lanzhou, 730050, Gansu, People's Republic of China.
Sci Rep. 2025 Jan 6;15(1):875. doi: 10.1038/s41598-025-85286-7.
4',5,6,7-tetrahydoxyisoflavone (6-hydroxygenistein, 6-OHG) is a hydroxylated derivative of genistein with excellent antioxidant activity, but whether 6-OHG can protect hypoxia-induced damage is unclear. The objective of current study was to evaluate the protective effect and underling mechanism of 6-OHG against hypoxia-induced injury via network pharmacology and cellular experiments. 6-OHG-related and hypoxia injury-related targets were screened by public databases. The intersected targets were used for constructing PPI network and performing GO and KEGG functional analysis. We induced injury in PC12 cells under hypoxia conditions and observed the effects and molecular mechanisms of 6-OHG on cellular damage. Network pharmacological analysis predicted that 6-OHG delayed hypoxia injury by mitigating oxidative stress, inflammatory response and apoptosis. Cellular experiments suggested that 6-OHG treatment mitigated cell damage, enhanced cell viability, reduced ROS production and MDA level, increased SOD and CAT activities and elevated GSH level in PC12 cell exposed to hypoxia. Additionally, 6-OHG treatment reduced the TNF-α and IL-6 levels and elevated the IL-10 content, while downregulated the NF-κB and TNF-α expressions. 6-OHG also inhibited the caspase-3 and - 9 activation and the Bax and cleaved caspase-3 expressions, and elevated the Bcl-2 expression. Moreover, 6-OHG remarkably enhanced Nrf2 nuclear translocation and increased HO-1 expression. Molecular docking also proved the strong binding affinities of 6-OHG with Nrf2 and HO-1. Furthermore, ML385, a specific Nrf2 inhibitor, eliminated the beneficial effects of 6-OHG. In summary, 6-OHG can alleviate hypoxia-induced injury in PC12 cells through activating Nrf2/HO-1 signaling pathway and may be developed as candidate for preventing neuro-damage induced by hypoxia.
4',5,6,7-四羟基异黄酮（6-羟基金雀异黄素，6-OHG）是金雀异黄素的一种羟基化衍生物，具有出色的抗氧化活性，但6-OHG是否能保护缺氧诱导的损伤尚不清楚。本研究的目的是通过网络药理学和细胞实验评估6-OHG对缺氧诱导损伤的保护作用及潜在机制。通过公共数据库筛选与6-OHG相关和与缺氧损伤相关的靶点。将交集靶点用于构建蛋白质-蛋白质相互作用（PPI）网络并进行基因本体论（GO）和京都基因与基因组百科全书（KEGG）功能分析。我们在缺氧条件下诱导PC12细胞损伤，并观察6-OHG对细胞损伤的影响及分子机制。网络药理学分析预测，6-OHG通过减轻氧化应激、炎症反应和细胞凋亡来延缓缺氧损伤。细胞实验表明，6-OHG处理减轻了缺氧暴露的PC12细胞的损伤，提高了细胞活力，降低了活性氧（ROS）生成和丙二醛（MDA）水平，增加了超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性，并提高了谷胱甘肽（GSH）水平。此外，6-OHG处理降低了肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）水平，提高了白细胞介素-10（IL-10）含量，同时下调了核因子-κB（NF-κB）和TNF-α表达。6-OHG还抑制了半胱天冬酶-3（caspase-3）和半胱天冬酶-9（caspase-9）的激活以及Bax和切割的caspase-3表达，并提高了Bcl-2表达。此外，6-OHG显著增强了核因子E2相关因子2（Nrf2）的核转位并增加了血红素加氧酶-1（HO-1）表达。分子对接也证明了6-OHG与Nrf2和HO-1具有很强的结合亲和力。此外，特异性Nrf2抑制剂ML385消除了6-OHG的有益作用。综上所述，6-OHG可通过激活Nrf2/HO-1信号通路减轻PC12细胞缺氧诱导的损伤，可能被开发为预防缺氧诱导神经损伤的候选药物。
