Ferrostatin-1 improves BMSC survival by inhibiting ferroptosis

Qiumin Xie; Yanting Sun; Hao Xu; Tangtian Chen; Han Xiang; Huiwen Liu; Rui Wang; Bin Tan; Qin Yi; Jie Tian; Jing Zhu

doi:10.1016/j.abb.2023.109535

Ferrostatin-1 improves BMSC survival by inhibiting ferroptosis

Arch Biochem Biophys. 2023 Mar 1:736:109535. doi: 10.1016/j.abb.2023.109535. Epub 2023 Jan 25.

Authors

Qiumin Xie¹, Yanting Sun¹, Hao Xu², Tangtian Chen¹, Han Xiang¹, Huiwen Liu¹, Rui Wang¹, Bin Tan¹, Qin Yi¹, Jie Tian³, Jing Zhu⁴

Affiliations

¹ Department of Pediatric Research Institute, Children's Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Key Laboratory of Pediatrics, Chongqing, 400014, China.
² Department of Clinical Laboratory, Children's Hospital of Chongqing Medical University, Chongqing, 400014, China.
³ Department of Cardiovascular (Internal Medicine), Children's Hospital of Chongqing Medical University, Chongqing, 400014, China.
⁴ Department of Pediatric Research Institute, Children's Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Key Laboratory of Pediatrics, Chongqing, 400014, China. Electronic address: jingzhu@cqmu.edu.cn.

PMID: 36708941
DOI: 10.1016/j.abb.2023.109535

Abstract

Objective: To investigate the effect of ferroptosis in BMSCs and explore the protective metabolism of ferrostatin-1 under GSDH treatment.

Methods: BMSCs 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.

Results: GSDH 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.

Conclusion: Ferr-1 rescued ferroptosis-induced injury to BMSCs under GSDH conditions, and AMPK might have a relationship with the mitigative effect of Ferr-1.

Keywords: AMPK; BMSCs; Ferroptosis; Ferrostatin-1; GSDH.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

AMP-Activated Protein Kinases*
Antioxidants
Ferroptosis*
Reactive Oxygen Species

Substances

ferrostatin-1
AMP-Activated Protein Kinases
Antioxidants
Reactive Oxygen Species