2. Academic Validation
  3. Inhibition of SLC40A1 represses osteoblast formation via inducing iron accumulation and activating the PERK/ATF4/CHOP pathway mediated oxidative stress

Inhibition of SLC40A1 represses osteoblast formation via inducing iron accumulation and activating the PERK/ATF4/CHOP pathway mediated oxidative stress

  • Redox Rep. 2024 Dec;29(1):2428147. doi: 10.1080/13510002.2024.2428147.
Yu Fang 1,  Wei Li 2,  Chongyang Dong 3,  Binli Gao 1,  Wen Guo 1,  Mingyu Li 1,  Zhichao Jiao 1
Affiliations

Affiliations

  • 1 Department of Orthopedics, Medical Affairs Department, The Affiliated Hospital of Inner Mongolia Medical University, Hohhot, People's Republic of China.
  • 2 Department of Radiology, The Second Affiliated Hospital of Inner Mongolia Medical University, Hohhot, People's Republic of China.
  • 3 College of Traditional Chinese Medicine, Inner Mongolia Medical University, Hohhot, People's Republic of China.
PMID: 39607819 DOI: 10.1080/13510002.2024.2428147
Abstract

Objective: This study aimed to investigate the effects of solute carrier family 40 member 1 (SLC40A1) on iron accumulation, oxidative stress and differentiation in osteoblasts and potential mechanisms.

Methods: Mouse preosteoblastic MC3T3-E1 cells were transfected with the SLC40A1 overexpression vector (oeSLC40A1) and siRNA (siSLC40A1), then cell differentiation was induced via ascorbic acid and β-glycerophosphate. Besides, Ferrostatin-1 (Ferroptosis inhibitor) and GSK2606414 (PERK Inhibitor) were added with siSLC40A1.

Results: Fe2+, malondialdehyde (MDA), and Reactive Oxygen Species (ROS) were higher but reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio was lower after siSLC40A1 transfection, while reduced Fe2+ and ROS but elevated GSH/GSSG ratio was observed after oeSLC40A1 transfection. Alkaline Phosphatase (ALP) staining, Alizarin Red S (ARS) staining, osteopontin (OPN) and Bone Morphogenetic Protein 2 (BMP2) were lower after siSLC40A1 transfection but were greater after oeSLC40A1 transfection. Furthermore, SLC40A1 negatively regulated the PERK/ATF4/CHOP pathway. Further exploration revealed that Fe2+, MDA, ROS, and the PERK/ATF4/CHOP pathway were attenuated, while GSH/GSSG ratio, ALP staining, ARS staining, and OPN expression were increased after ferrostatin-1 treatment in the siSLC40A1-transfected cells. Similar trends were observed with respect to GSK2606414 treatment with siSLC40A1.

Conclusion: SLC40A1 inhibition suppresses osteoblast formation by facilitating iron accumulation and activating the PERK/ATF4/CHOP pathway-mediated oxidative stress.

Keywords

GSK2606414; PERK/ATF4/CHOP pathway; SLC40A1; ferrostatin-1; iron accumulation; osteoblast differentiation; osteoblast formation; oxidative stress.

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