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Ferroptosis inhibitor liproxstatin-1 alleviates metabolic dysfunction-associated fatty liver disease in mice: potential involvement of PANoptosis

Acta Pharmacologica Sinica (2022)Cite this article

Abstract

Ferroptosis is a new form of regulated cell death characterized by excessive iron accumulation and uncontrollable lipid peroxidation. The role of ferroptosis in metabolic dysfunction-associated fatty liver disease (MAFLD) is not fully elucidated. In this study we compared the therapeutic effects of ferroptosis inhibitor liproxstatin-1 (LPT1) and iron chelator deferiprone (DFP) in MAFLD mouse models. This model was established in mice by feeding a high-fat diet with 30% fructose in water (HFHF) for 16 weeks. The mice then received LPT1 (10 mg·kg−1·d−1, ip) or DFP (100 mg·kg−1·d−1, ig) for another 2 weeks. We showed that both LPT1 and DFP treatment blocked the ferroptosis markers ACSL4 and ALOX15 in MAFLD mice. Furthermore, LPT1 treatment significantly reduced the liver levels of triglycerides and cholesterol, lipid peroxidation markers 4-hydroxynonenal (4-HNE) and malondialdehyde (MDA), and ameliorated the expression of lipid synthesis/oxidation genes (Pparα, Scd1, Fasn, Hmgcr and Cpt1a), insulin resistance, mitochondrial ROS content and liver fibrosis. Importantly, LPT1 treatment potently inhibited hepatic apoptosis (Bax/Bcl-xL ratio and TUNEL+ cell number), pyroptosis (cleavages of Caspase-1 and GSDMD) and necroptosis (phosphorylation of MLKL). Moreover, LPT1 treatment markedly inhibited cleavages of PANoptosis-related caspase-8 and caspase-6 in MAFLD mouse liver. In an in vitro MAFLD model, treatment with LPT1 (100 nM) prevented cultured hepatocyte against cell death induced by pro-PANoptosis molecules (TNF-α, LPS and nigericin) upon lipid stress. On the contrary, DFP treatment only mildly attenuated hepatic inflammation but failed to alleviate lipid deposition, insulin resistance, apoptosis, pyroptosis and necroptosis in MAFLD mice. We conclude that ferroptosis inhibitor LPT1 protects against steatosis and steatohepatitis in MAFLD mice, which may involve regulation of PANoptosis, a coordinated cell death pathway that involves apoptosis, pyroptosis and necroptosis. These results suggest a potential link between ferroptosis and PANoptosis.

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Fig. 1: The inhibitor LPT1 blocks ferroptosis without disrupting iron accumulation in a mouse MAFLD model.
Fig. 2: The ferroptosis inhibitor LPT1 decreases steatosis and mitochondrial ROS in a mouse MAFLD model.
Fig. 3: The ferroptosis inhibitor LPT1 mitigates liver injury and fibrosis in a mouse MAFLD models.
Fig. 4: The ferroptosis inhibitor LPT1 suppresses steatohepatitis and ‘PANoptosis’ in a mouse MAFLD models.
Fig. 5: The ferroptosis inhibitor LPT1 inhibits cleavage of PANoptosis-related Caspase-6 and Caspase-8.
Fig. 6: The ferroptosis inhibitor LPT1 directly blocks PANoptosis in hepatocytes upon lipid stress.
Fig. 7: An iron chelator DFP reduces ferroptosis but does not ameliorate steatosis.
Fig. 8: An iron chelator DFP alleviates steatohepatitis.
Fig. 9: An iron chelator DFP does not block PANoptosis.

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Funding

This work was supported by the grants from National Natural Science Foundation (82073915, 82274030, 81971306, and 81773719), National Key Research and Development Program Stem Cell and Translational Research Key Projects (2018YFA0108301), Shanghai Talent Development Fund (2020091), Shanghai Science and Technology Commission Grants (21XD1424900, 2020-JMRH1-KJ12, 21S11901200 and 19140904700), Shanghai Shuguang Program (19SG32) and Shanghai Rising Stars of Medical Talent Development Program-Clinical Pharmacist Project [SHWRS(2020)_087].

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  1. These authors contributed equally: Jie Tong, Xiu-ting Lan, Zhen Zhang

Authors and Affiliations

  1. Department of Pharmacy, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, 200072, China

    Jie Tong, Xiu-ting Lan, Zhen Zhang, Yi Liu, Xu-jie Wang, Shen-xi Ou-Yang, Fu-ming Shen & Dong-jie Li

  2. School of Pharmacy, Naval Medical University/Second Military Medical University, Shanghai, 200433, China

    Di-yang Sun, Chun-lin Zhuang & Pei Wang

  3. Institute of Nuclear Medicine, Tongji University School of Medicine, Shanghai, 200072, China

    Dong-jie Li

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  1. Jie Tong
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Contributions

JT: Writing-original draft, Methodology, Investigation. XTL: Methodology, Formal analysis, Investigation. ZZ: Formal analysis and Investigation. YL: Formal analysis, Investigation, Writing – review & editing. DYS: Formal analysis and Writing – review & editing. XJW: Investigation and Writing – review & editing. SXOY: Investigation and Writing – review & editing. CLZ: Resources (Synthesis of LPT1). FMS: Formal analysis, Resources and Writing – review & editing. PW: Conceptualization, Writing – original draft, Supervision, Funding acquisition, Writing-review & editing. DJL: Conceptualization, Writing – original draft, Supervision, Funding acquisition, Writing – review & editing.

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Correspondence to Pei Wang or Dong-jie Li.

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Tong, J., Lan, Xt., Zhang, Z. et al. Ferroptosis inhibitor liproxstatin-1 alleviates metabolic dysfunction-associated fatty liver disease in mice: potential involvement of PANoptosis. Acta Pharmacol Sin (2022). https://doi.org/10.1038/s41401-022-01010-5

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Keywords

  • ferroptosis
  • PANoptosis
  • MAFLD
  • liver disease

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