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Endothelial Gsα deficiency promotes ferroptosis and exacerbates atherosclerosis in apolipoprotein E-deficient mice via the inhibition of NRF2 signaling

Acta Pharmacologica Sinica volume 46, pages 1289–1302 (2025)Cite this article

Abstract

The importance of ferroptosis in the occurrence and progression of atherosclerosis is gradually being recognized. The stimulatory G protein α subunit (Gsα) plays a crucial role in the physiology of endothelial cells (ECs). Our previous study showed that endothelial Gsα could regulate angiogenesis and preserve endothelial permeability. In this study, we investigated whether endothelial Gsα contributed to atherosclerosis through ferroptosis and oxidative stress. We generated endothelial Gsα-specific knockout mice in apolipoprotein E-deficient (ApoE−/−) background (ApoE−/−GsαECKO), and found that the mice exhibited aggravated atherosclerotic lesions and signs of ferroptosis compared with their wild-type littermates (ApoE−/−Gsαfl/fl). In human aortic endothelial cells (HAECs), overexpression of Gsα reduced lipid peroxidation and ferroptosis, whereas Gsα knockdown exacerbated oxidative stress and ferroptosis. Further, Gsα overexpression in HAECs increased the expression of antioxidant genes nuclear factor erythroid 2-related 2 (NRF2) and its downstream genes. Gsα regulated the expression of NRF2 through CCCTC-binding factor (CTCF). In conclusion, this study has revealed that Gsα acts as a defense factor against endothelial ferroptosis and is a potential target for the treatment of atherosclerosis and associated ischemic heart disease.

A model depicting the increase in the endothelial Gsα protein level in response to atherosclerotic stimuli. Gsα regulates NRF2 expression through cAMP/Epac/CTCF-mediated transcription and inhibits ferroptosis. Endothelial Gsα deficiency alleviates antioxidative stress and exacerbates atherosclerosis.

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Fig. 1: Endothelial Gsα expression is increased in atherosclerosis.
Fig. 2: Genetic deficiency of Gsα aggravates the progression of atherosclerosis in ApoE−/− mice.
Fig. 3: Gsα inhibits vascular endothelial injury by attenuating the ferroptosis pathway.
Fig. 4: Gsα deficiency inhibits NRF2 signaling in endothelial cells.
Fig. 5: Gsα overexpression enhances the NRF2 signaling pathway in endothelial cells.
Fig. 6: Gsα inhibits the ferroptosis pathway via NRF2.
Fig. 7: Endothelial Gsα regulates NRF2 via CTCF.
Fig. 8: Endothelial Gsα regulates CTCF via the cAMP/Epac signaling pathway.

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Acknowledgements

The authors thank Prof. Yulong He from Soochow University for providing the Cdh5-CreERT2 mice.

Funding

This study was supported by grants from the Taishan Scholar Project of Shandong Province of China (No. tstp20240852), the Natural Science Foundation of Shandong Provincial (ZR2021QH112, ZR2024ZD23), the Natural Science Foundation for Distinguished Young Scolars of Shandong Province (ZR2020JQ30), the National Natural Science Foundation of China (No. 82100471, 82470499, 82270457), and the Postdoctoral Innovation Project of Shandong Province (SDCX-ZG-202400016).

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  1. These authors contributed equally: Li-fan He, Lei Wang

Authors and Affiliations

  1. State Key Laboratory for Innovation and Transformation of Luobing Theory; Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education, Chinese National Health Commission and Chinese Academy of Medical Sciences; Department of Cardiology, Qilu Hospital of Shandong University, Jinan, 250012, China

    Li-fan He, Lei Wang, Jing-wei Li, Xiao Xiong, Xiao-lin Yue, Pei-dong Yuan, Han-lin Lu, Fang-pu Yu, Jian-min Yang, Cheng Zhang, Xiaoteng Qin & Wencheng Zhang

  2. Department of Critical Care Medicine, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, 250012, China

    Li-fan He & Lei Wang

  3. School of Life Science and Key Laboratory of the Ministry of Education for Experimental Teratology, Shandong University, Jinan, 250013, China

    Jian-gang Gao

  4. Metabolic Diseases Branch, National Institute of Diabetes, Digestive, and Kidney Diseases, National Institutes of Health, Bethesda, MD, 20814, USA

    Min Chen & Lee S. Weinstein

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Contributions

L-FH, LW, XTQ, WCZ, and CZ. designed the study. LFH, LW, J-WL, H-LL, XX, X-LY, P-DY, and J-GG, performed the experiments and analyzed the results. LFH, and WCZ drafted the manuscript. MC, F-PY, LSW, J-MY, JGG, CZ, and WCZ revised the manuscript for important intellectual content. All the authors have read and approved the final manuscript.

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Correspondence to Xiaoteng Qin or Wencheng Zhang.

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He, Lf., Wang, L., Li, Jw. et al. Endothelial Gsα deficiency promotes ferroptosis and exacerbates atherosclerosis in apolipoprotein E-deficient mice via the inhibition of NRF2 signaling. Acta Pharmacol Sin 46, 1289–1302 (2025). https://doi.org/10.1038/s41401-024-01446-x

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