PDF(Pubmed)
Abstract：
Neuronal precursor cells expressed developmentally down-regulated 4 (Nedd4) are believed to play a critical role in promoting the degradation of substrate proteins and are involved in numerous biological processes. However, the role of Nedd4 in intracerebral hemorrhage (ICH) remains unknown. This study aims to investigate the regulatory role of Nedd4 in the ICH model.
Male C57BL/6J mice were induced with ICH. Subsequently, the levels of glutathione peroxidase 4 (GPX4), malondialdehyde (MDA) concentration, iron content, mitochondrial morphology, as well as the expression of divalent metal transporter 1 (DMT1) and Nedd4 were assessed after ICH. Furthermore, the impact of Nedd4 overexpression was evaluated through analyses of hematoma area, ferroptosis, and neurobehavioral function. The mechanism underlying Nedd4-mediated degradation of DMT1 was elecidated using immunoprecipitation (IP) after ICH.
Upon ICH, the level of DMT1 in the brain increased, but decreased when Nedd4 was overexpressed using Lentivirus, suggesting a negative correlation between Nedd4 and DMT1. Additionally, the degradation of DMT1 was inhibited after ICH. Furthermore, it was found that Nedd4 can interact with and ubiquitinate DMT1 at lysine residues 6, 69, and 277, facilitating the degradation of DMT1. Functional analysis indicated that overexpression of Nedd4 can alleviate ferroptosis and promote recovery following ICH.
The results demonstrated that ferroptosis occurs via the Nedd4/DMT1 pathway during ICH, suggesting it potential as a valuable target to inhibit ferroptosis for the treatment of ICH.
Male C57BL/6J mice were induced with ICH. Subsequently, the levels of glutathione peroxidase 4 (GPX4), malondialdehyde (MDA) concentration, iron content, mitochondrial morphology, as well as the expression of divalent metal transporter 1 (DMT1) and Nedd4 were assessed after ICH. Furthermore, the impact of Nedd4 overexpression was evaluated through analyses of hematoma area, ferroptosis, and neurobehavioral function. The mechanism underlying Nedd4-mediated degradation of DMT1 was elecidated using immunoprecipitation (IP) after ICH.
Upon ICH, the level of DMT1 in the brain increased, but decreased when Nedd4 was overexpressed using Lentivirus, suggesting a negative correlation between Nedd4 and DMT1. Additionally, the degradation of DMT1 was inhibited after ICH. Furthermore, it was found that Nedd4 can interact with and ubiquitinate DMT1 at lysine residues 6, 69, and 277, facilitating the degradation of DMT1. Functional analysis indicated that overexpression of Nedd4 can alleviate ferroptosis and promote recovery following ICH.
The results demonstrated that ferroptosis occurs via the Nedd4/DMT1 pathway during ICH, suggesting it potential as a valuable target to inhibit ferroptosis for the treatment of ICH.
摘要：
目的：神经元前体细胞表达下调4（Nedd4）被认为在促进底物蛋白降解中起关键作用，并参与许多生物学过程。然而,Nedd4在脑出血(ICH)中的作用尚不清楚.本研究旨在探讨Nedd4在ICH模型中的调节作用。
方法:用ICH诱导雄性C57BL/6J小鼠。随后，谷胱甘肽过氧化物酶4（GPX4）的水平，丙二醛（MDA）浓度，铁含量,线粒体形态学，以及在ICH后评估二价金属转运蛋白1（DMT1）和Nedd4的表达。此外，Nedd4过表达的影响通过分析血肿面积，铁性凋亡,和神经行为功能。在ICH后使用免疫沉淀（IP）确定了Nedd4介导的DMT1降解的潜在机制。
结果：在ICH时，大脑中的DMT1水平升高，但是当使用慢病毒过度表达Nedd4时，表明Nedd4和DMT1之间呈负相关。此外,ICH后DMT1的降解受到抑制。此外，发现Nedd4可以在赖氨酸残基6、69和277处与DMT1相互作用并使其泛素化，从而促进DMT1的降解。功能分析表明，Nedd4的过表达可以减轻ICH后的铁凋亡并促进其恢复。
结论：结果表明，在ICH期间通过Nedd4/DMT1途径发生铁凋亡，这表明它有可能作为一个有价值的靶点来抑制铁蛋白凋亡治疗ICH。
方法:用ICH诱导雄性C57BL/6J小鼠。随后，谷胱甘肽过氧化物酶4（GPX4）的水平，丙二醛（MDA）浓度，铁含量,线粒体形态学，以及在ICH后评估二价金属转运蛋白1（DMT1）和Nedd4的表达。此外，Nedd4过表达的影响通过分析血肿面积，铁性凋亡,和神经行为功能。在ICH后使用免疫沉淀（IP）确定了Nedd4介导的DMT1降解的潜在机制。
结果：在ICH时，大脑中的DMT1水平升高，但是当使用慢病毒过度表达Nedd4时，表明Nedd4和DMT1之间呈负相关。此外,ICH后DMT1的降解受到抑制。此外，发现Nedd4可以在赖氨酸残基6、69和277处与DMT1相互作用并使其泛素化，从而促进DMT1的降解。功能分析表明，Nedd4的过表达可以减轻ICH后的铁凋亡并促进其恢复。
结论：结果表明，在ICH期间通过Nedd4/DMT1途径发生铁凋亡，这表明它有可能作为一个有价值的靶点来抑制铁蛋白凋亡治疗ICH。
