Ferrostatin-1 improves BMSC survival by inhibiting ferroptosis
Ferrostatin-1 improves BMSC survival by inhibiting ferroptosis
复制标题
DOI：
10.1016/j.abb.2023.109535
复制
发表时间：
2023-02-04
影响因子：
3.9
通讯作者：
Zhu，Jing
中科院分区：
文献类型：
--
作者：
Xie，Qiumin;Sun，Yanting;Zhu，Jing
目的观察GSDH对骨髓间充质干细胞铁凋亡的影响，探讨其铁抑制素-1的保护性代谢作用。方法用GSDH处理骨髓间充质干细胞，模拟体内损伤微环境，建立细胞损伤模型。用碘化丙啶和CCK8检测死细胞率和细胞活力。DCFH-DA和Amplex Red、FerroOrange和BPDIPY分别用于显示ROS、Fe2+和脂滴的细胞荧光图像。脂质过氧化MDA测定试剂盒定量检测MDA。采用JC-1染色、Mito-Tracker染色和TEM分别检测线粒体的膜电位、形态和超微结构。Western blotting检测凋亡相关蛋白GPX4、FTH1的表达水平。结果gsdh诱导骨髓间充质干细胞铁凋亡的基础是细胞死亡率、细胞中Fe2+、ROS、脂滴和MDA的增加，以及抗氧化系统（如GPX4）蛋白水平的降低，以及脂肪酸合成相关蛋白水平的增加。与空白组相比，GSDH组线粒体膜电位降低，形态受损，超微结构萎缩；Ferr-1在一定程度上挽救了损伤的骨髓间充质干细胞，降低了细胞死亡率、Fe2+、ROS、脂滴、MDA，提高了抗氧化蛋白水平。fer1处理后AMPK被磷酸化活化，其下游脂质过氧化和抗氧化蛋白发生相应变化。抑制AMPK抑制了Ferr-1的疗效。结论铁-1可挽救GSDH条件下铁凋亡诱导的骨髓间充质干细胞损伤，AMPK可能与铁-1的缓解作用有关。
ObjectiveTo investigate the effect of ferroptosis in BMSCs and explore the protective metabolism of ferrostatin-1 under GSDH treatment.MethodsBMSCs were treated with GSDH to simulate the damaged microenvironment in vivo to establish a cell injury model. Propidium iodide and CCK8 were utilized to detect the ratio of dead cells and cell viability. DCFH-DA and Amplex Red, FerroOrange, and BPDIPY were used to visualize the cellular fluorescent images of ROS, Fe2+, and lipid droplets, respectively. The quantified detection of MDA was conducted by a Lipid Peroxidation MDA Assay Kit. JC-1 staining, Mito-Tracker staining, and TEM were implemented to detect the membrane potential, morphology, and ultrastructure of mitochondria, respectively. The expression levels of ferroptosis-related proteins such as GPX4 and FTH1 were measured by Western blotting.ResultsGSDH treatment induced ferroptosis in BMSCs based on an increased ratio of cell death, Fe2+, ROS, lipid droplets, and MDA in cells plus decreased protein levels of antioxidant systems, such as GPX4, and increased protein levels related to fatty acid synthesis. Compared to the blank group, mitochondria in the GSDH group underwent lower membrane potential, damaged morphology, and shrunken ultrastructure; Ferr-1 rescued the injured BMSCs to a certain extent as the declined ratio of cell death, Fe2+, ROS, lipid droplets, MDA, and the increased level antioxidant protein. AMPK was phosphorylated and activated after Ferr-1 treatment, and its downstream lipid peroxidation and antioxidation proteins changed accordingly. Inhibition of AMPK hindered the curative effect of Ferr-1.ConclusionFerr-1 rescued ferroptosis-induced injury to BMSCs under GSDH conditions, and AMPK might have a relationship with the mitigative effect of Ferr-1.