Wen Huizhen, Tu Xingxing, Luo Fulan, Zeng Chunyuan, Xia Chuang, Zhao Qian, Xian Zihong, Zhou Zhongzhen, Xu Jiangping, Wang Haitao
NMPA Key Laboratory for Research and Evaluation of Drug Metabolism & Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, 510515, China.
NMPA Key Laboratory for Research and Evaluation of Drug Metabolism & Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, 510515, China; Key Laboratory of Mental Health of the Ministry of Education, Southern Medical University, Guangzhou, 510515, China; Center for Brain Science and Brain-Inspired Intelligence, Guangdong-Hong Kong-Macao Greater Bay Area, Guangzhou, 510515, China.
Chem Biol Interact. 2025 Feb 25;408:111405. doi: 10.1016/j.cbi.2025.111405. Epub 2025 Jan 29.
Dynamin-related protein 1 (Drp1) regulates mitochondrial fission and participates in neuronal apoptosis during the pathology of cerebral ischemia. We have previously shown that inhibition of phosphodiesterase-4 (PDE4) protects against neuronal apoptosis in models of ischemic stroke. However, it remains unclear whether PDE4 inhibition affects Drp1-mediated mitochondrial dysfunction and apoptosis under cerebral ischemia conditions. This study aimed to determine whether ZX21011, a novel PDE4 inhibitor synthesized in our laboratory, can act on Drp1 to counteract ischemic brain injury and to elucidate its mechanism of action. We demonstrated that ZX21011 effectively reduced neuronal apoptosis caused by oxygen-glucose deprivation/reoxygenation (OGD/R) in HT22 cells and ameliorated neurological deficits caused by middle cerebral artery occlusion/reperfusion (MCAO/R) in rats. ZX21011 enhanced glycogen synthase kinase-3β (GSK3β) phosphorylation (Ser9), GSK3β(S9A) mutation blocked the protective effects of ZX21011. Simultaneously, ZX21011 reduced the levels of reactive oxygen species (ROS), restored the morphology of mitochondria, and inhibited the phosphorylation of Drp1(Ser616). The Drp1(S616E) mutation blocked the protective effects of ZX21011 on ROS production and mitochondrial morphology function after cerebral ischemia. What's more, co-immunoprecipitation analysis revealed that ZX21011 decreased the binding of GSK3β to Drp1, and GSK3β(S9A) mutation reversed the effects of ZX21011 on Drp1 phosphorylation and cell viability. Moreover, ZX21011 decreased Drp1(Ser616) phosphorylation within the ischemic penumbra of rats following cerebral ischemia/reperfusion. In summary, ZX21011 counteracts ischemic stroke-induced oxidative stress and neuronal death, and its action is related to decreased Drp1 phosphorylation at Ser616. Thus, ZX21011 is a potential compound for the intervention of stroke.
动力相关蛋白1（Drp1）在脑缺血病理过程中调节线粒体分裂并参与神经元凋亡。我们之前已经表明，在缺血性中风模型中，抑制磷酸二酯酶4（PDE4）可防止神经元凋亡。然而，尚不清楚在脑缺血条件下，PDE4抑制是否会影响Drp1介导的线粒体功能障碍和凋亡。本研究旨在确定我们实验室合成的新型PDE4抑制剂ZX21011是否能作用于Drp1以对抗缺血性脑损伤并阐明其作用机制。我们证明，ZX21011有效减少了HT22细胞中氧糖剥夺/复氧（OGD/R）引起的神经元凋亡，并改善了大鼠大脑中动脉闭塞/再灌注（MCAO/R）引起的神经功能缺损。ZX21011增强了糖原合酶激酶-3β（GSK3β）的磷酸化（Ser9），GSK3β（S9A）突变阻断了ZX21011的保护作用。同时，ZX21011降低了活性氧（ROS）水平，恢复了线粒体形态，并抑制了Drp1（Ser616）的磷酸化。Drp1（S616E）突变阻断了ZX21011对脑缺血后ROS产生和线粒体形态功能的保护作用。此外，免疫共沉淀分析表明，ZX21011降低了GSK3β与Drp1的结合，GSK3β（S9A）突变逆转了ZX21011对Drp1磷酸化和细胞活力的影响。此外，ZX21011降低了脑缺血/再灌注后大鼠缺血半暗带内Drp1（Ser616）的磷酸化。总之，ZX21011可对抗缺血性中风诱导的氧化应激和神经元死亡，其作用与降低Ser616处的Drp1磷酸化有关。因此，ZX21011是一种潜在的中风干预化合物。
