Tocopherol attenuates the oxidative stress of BMSCs by inhibiting ferroptosis through the PI3k/AKT/mTOR pathway

Dongmei Lan; Chao Yao; Xue Li; Haijiang Liu; Dan Wang; Yan Wang; Shengcai Qi

doi:10.3389/fbioe.2022.938520

Tocopherol attenuates the oxidative stress of BMSCs by inhibiting ferroptosis through the PI3k/AKT/mTOR pathway

Front Bioeng Biotechnol. 2022 Aug 17:10:938520. doi: 10.3389/fbioe.2022.938520. eCollection 2022.

Authors

Dongmei Lan^{1

2

3}, Chao Yao^{1

2

3}, Xue Li^{1

2

3}, Haijiang Liu^{2

4}, Dan Wang^{5

6}, Yan Wang^{2

7}, Shengcai Qi^{1

2

3}

Affiliations

¹ Department of Prosthodontics, Shanghai Stomatological Hospital & School of Stomatology, Fudan University, Shanghai, China.
² Shanghai Key Laboratory of Craniomaxillofacial Development and Diseases, Fudan University, Shanghai, China.
³ Medical College, Anhui University of Science and Technology, Huainan, China.
⁴ Department of Endodontics, Shanghai Stomatological Hospital & School of Stomatology, Fudan University, Shanghai, China.
⁵ Institute for Tissue Engineering and Regenerative Medicine, School of Biomedical Sciences, Ministry of Education Key Laboratory for Regenerative Medicine, Department of Orthopaedics and Traumatology, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, Hong Kong SAR, China.
⁶ Center for Neuromusculoskeletal Restorative Medicine, Hong Kong Science Park, Hong Kong, Hong Kong SAR, China.
⁷ Department of Preventive Dentistry, Shanghai Stomatological Hospital & School of Stomatology, Fudan University, Shanghai, China.

Abstract

Oxidative stress can induce bone tissue damage and the occurrence of multiple diseases. As a type of traditional medicine, tocopherol has been reported to have a strong antioxidant effect and contributes to osteogenic differentiation. The purpose of this study was to investigate the protective effect of tocopherol on the oxidative stress of rat bone marrow-derived mesenchymal stem cells (BMSCs) and the underlying mechanisms. By establishing an oxidative stress model in vitro, the cell counting kit-8 (CCK-8), reactive oxygen species (ROS) analysis, Western blot (WB), real-time PCR (RT-PCR), alkaline phosphatase (ALP) staining, and Alizarin Red staining (ARS) evaluated the effects of tocopherol on the cell viability, intracellular ROS levels, and osteogenic differentiation in BMSCs. In addition, ferroptosis-related markers were examined via Western blot, RT-PCR, and Mito-FerroGreen. Eventually, the PI3K/AKT/mTOR signaling pathway was explored. We found that tocopherol significantly maintained the cell viability, reduced intracellular ROS levels, upregulated the levels of anti-oxidative genes, promoted the levels of osteogenic-related proteins, and the mRNA of BMSCs stimulated by H₂O₂. More importantly, tocopherol inhibited ferroptosis and upregulated the phosphorylation levels of PI3K, AKT, and mTOR of BMSCs upon H₂O₂ stimulation. In summary, tocopherol protected BMSCs from oxidative stress damage via the inhibition of ferroptosis through the PI3K/AKT/mTOR pathway.

Keywords: BMSCs; PI3K/Akt/mTOR pathway; ferroptosis; osteogenic differentiation; tocopherol.

Grants and funding

This research was funded by: 1) Science and Technology Commission of Shanghai Municipality, grant numbers: 20ZR1443100 and 21140904500; 2) National Natural Science Foundation of China, grant number: 81771035.