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Genipin protects against acute liver injury by abrogating ferroptosis via modification of GPX4 and ALOX15-launched lipid peroxidation in mice

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Abstract

It is essential to further characterize liver injury aimed at developing novel therapeutic approaches. This study investigated the mechanistic basis of genipin against carbon tetrachloride (CCl4)-triggered acute liver injury concerning ferroptosis, a novel discovered modality of regulated cell death. All experiments were performed using hepatotoxic models upon CCl4 exposure in mice and human hepatocytes in vitro. Immunohistochemistry, immunoblotting, molecular docking, RNA-sequencing and ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) were conducted. CCl4 intoxication was manifested with lipid peroxidation-dictated ferroptotic cell death, together with changes in a cascade of ferroptosis-associated events and several regulatory pathways. Both the administration of genipin and ferrostatin-1 (Fer-1) significantly prevented this hepatotoxicity in response to CCl4 intoxication via upregulating GPX4 and xCT (i.e., critical regulators of ferroptosis). RNA-sequencing unraveled that arachidonic acid metabolism was considerably influenced upon genipin treatment. Accordingly, genipin treatment attenuated arachidonate 15-lipoxygenase (ALOX15)-launched lipid peroxidation in terms of UHPLC-MS/MS analysis and inflammation. In vitro, genipin supplementation rescued erastin-induced hepatocellular inviability and lipid ROS accumulation. The siRNA knockdown of GPX4 partially abrogated the protective effects of genipin on erastin-induced cytotoxicity, whereas the cytotoxicity was less severe in the presence of diminished ALOX15 expression in L-O2 cells. In conclusion, our findings uncovered that genipin treatment protects against CCl4-triggered acute liver injury by abrogating hepatocyte ferroptosis, wherein the pharmacological modification of dysregulated GPX4 and ALOX15-launched lipid peroxidation was responsible for underlying medicinal effects as molecular basis.

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Data Availability

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

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    Authors and Affiliations

    1. Department of Gastroenterology and Hepatology, Tianjin Medical University General Hospital, Anshan Road 154, Heping District, Tianjin, 300052, China

      Xiaofei Fan, Xiaoyu Wang, Yangyang Hui, Lihong Mao, Binxin Cui, Weilong Zhong & Chao Sun

    2. Tianjin Institute of Digestive Disease, Tianjin Medical University General Hospital, Anshan Road 154, Heping District, Tianjin, 300052, China

      Xiaofei Fan, Xiaoyu Wang, Yangyang Hui, Lihong Mao, Weilong Zhong & Chao Sun

    3. Department of Gastroenterology, Nanjing Drum Tower Hospital, Chinese Academy of Medical Science & Peking Union Medical College, Zhongshan Road 321,Gulou District, Nanjing, 210008, Jiangsu, China

      Tianming Zhao

    4. Department of Gastroenterology, Tianjin Medical University General Hospital Airport Hospital, Tianjin Airport Economic Area, East Street 6, Tianjin, 300308, China

      Binxin Cui & Chao Sun

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    Contributions

    Conception or design of the work: Xiaofei Fan, Xiaoyu Wang, Yangyang Hui and Chao Sun; Data collection: Tianming Zhao, Lihong Mao, Binxin Cui and Weilong Zhong; Drafting the article and critical revision of the article: Chao Sun; Final ap-proval of the version to be published: all authors.

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    Correspondence to Chao Sun.

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    All animal studies were approved by the Institutional Animal Care and Use Committee at Tianjin Medical University General Hospital (IRB2021-DWFL-142).

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    Xiaofei Fan, Xiaoyu Wang and Yangyang Hui contributed equally to this work.

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    Fan, X., Wang, X., Hui, Y. et al. Genipin protects against acute liver injury by abrogating ferroptosis via modification of GPX4 and ALOX15-launched lipid peroxidation in mice. Apoptosis 28, 1469–1483 (2023). https://doi.org/10.1007/s10495-023-01867-9

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