Genipin induces liver injury by disrupting the CPS1-regulated urea cycle and redox homeostasis

Hongzhan Xu; Xinhui Huang; Yujia Zhang; Shilin Gong; Yusha Luo; Yichao Fang; Xiaolan Hu; Miaomiao Luo; Yujia Zhai; Fangyuan Gao; Xinglong Chen; Rongping Zhang; Longshan Zhao; Yong Wang; Jianlin Wu; Xin Zhang; Huiqing Liang; Jun Wen; Tingting Zhou

doi:10.1016/j.jpha.2026.101647

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Hongzhan Xu, Xinhui Huang, Yujia Zhang, Shilin Gong, Yusha Luo, Yichao Fang, Xiaolan Hu, Miaomiao Luo, Yujia Zhai, Fangyuan Gao, Xinglong Chen, Rongping Zhang, Longshan Zhao, Yong Wang, Jianlin Wu, Xin Zhang, Huiqing Liang, Jun Wen, Tingting Zhou. Genipin induces liver injury by disrupting the CPS1-regulated urea cycle and redox homeostasis[J]. Journal of Pharmaceutical Analysis. doi: 10.1016/j.jpha.2026.101647

Citation:

Hongzhan Xu, Xinhui Huang, Yujia Zhang, Shilin Gong, Yusha Luo, Yichao Fang, Xiaolan Hu, Miaomiao Luo, Yujia Zhai, Fangyuan Gao, Xinglong Chen, Rongping Zhang, Longshan Zhao, Yong Wang, Jianlin Wu, Xin Zhang, Huiqing Liang, Jun Wen, Tingting Zhou. Genipin induces liver injury by disrupting the CPS1-regulated urea cycle and redox homeostasis[J]. Journal of Pharmaceutical Analysis. doi: 10.1016/j.jpha.2026.101647

Citation:

Hongzhan Xu, Xinhui Huang, Yujia Zhang, Shilin Gong, Yusha Luo, Yichao Fang, Xiaolan Hu, Miaomiao Luo, Yujia Zhai, Fangyuan Gao, Xinglong Chen, Rongping Zhang, Longshan Zhao, Yong Wang, Jianlin Wu, Xin Zhang, Huiqing Liang, Jun Wen, Tingting Zhou. Genipin induces liver injury by disrupting the CPS1-regulated urea cycle and redox homeostasis[J]. Journal of Pharmaceutical Analysis. doi: 10.1016/j.jpha.2026.101647

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Genipin induces liver injury by disrupting the CPS1-regulated urea cycle and redox homeostasis

doi: 10.1016/j.jpha.2026.101647

a. School of Pharmacy, Second Military Medical University, Shanghai, 200433, China;
b. Shanghai Key Laboratory for Pharmaceutical Metabolite Research, School of Pharmacy, Second Military Medical University, Shanghai, 200433, China;
c. Department of Pharmacy, The First Navy Hospital of Southern Theater Command, Zhanjiang, Guangdong, 524005, China;
d. Institute of Mass Spectrometry and Atmospheric Environment, Guangdong Provincial Key Laboratory of Speed Capability Research, Jinan University, Guangzhou, 510632, China;
e. Department of Pharmacy, Shanghai Xuhui District Central Hospital, Shanghai, 200020, China;
f. Naval Medical Center, Second Military Medical University, Shanghai, 200433, China;
g. Department of Health Toxicology, Faculty of Naval Medicine, Naval Medical University, Shanghai, 200433,China;
h. School of Chinese Materia Medica and Yunnan Key Laboratory of Southern Medicinal Resources, Yunnan University of Traditional Chinese Medicine, Kunming, 650500, China;
i. School of Pharmacy, Shenyang Pharaceutical University, Shenyang, 117004, China;
j. School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, 100029, China;
k. State Key Laboratory for Quality Research of Chinese Medicines, Macau University of Science and Technology, Taipa, Macau SAR, 999078, China;
l. Department of Gastroenterology, Changzheng Hospital, Naval Medical University, Shanghai, 200003,China;
m. Hepatology Unit, Xiamen Hospital of Traditional Chinese Medicine, Xiamen, Fujian, 361015, China

Funds:

This work was supported by the National Natural Science Foundation of China (Grant No.: 82474056), the Funds for International Cooperation and Exchange of the National Natural Science Foundation of China (Grant No.: 82461160264), the National Natural Science Foundation of China (Grant Nos.: 82404854 and 82304687), Noncommunicable Chronic Diseases-National Science and Technology Major Project, China (Grant No.: 2023ZD0502605), Shanghai Municipal Health Commission, China (Grant No.: 2022XD037), Science and Technology Development Fund, Macau SAR, China (Grant No.: FDCT0025/2021/A1), and Shanghai Magnolia Talent Plan Pujiang Project, China (Grant No.: 23PJD113). Graphical Abstract was created with BioRender.com.

Received Date: Jan. 15, 2026
Accepted Date: May 05, 2026
Rev Recd Date: May 05, 2026
Available Online: May 07, 2026

Abstract

Abstract

Genipin (GP) is the key metabolite of geniposide (GE), the primary active component of the traditional Chinese medicine Gardeniae Fructus. Its hepatotoxicity poses a potential risk to safe clinical administration, while its core pathophysiological mechanism remains unclear. This study systematically elucidated the molecular mechanisms of GP-induced hepatotoxicity through integrated multi-omics and functional validation. GP treatment induced significant liver damage and oxidative stress in rats, characterized by elevated serum transaminases (alanine aminotransferase (ALT) increased by approximately 63.5-fold; aspartate aminotransferase (AST) by 18.6-fold), an approximately 1.8-fold decrease in superoxide dismutase (SOD) activity, and an approximately 10-fold accumulation of malondialdehyde (MDA) (all P < 0.001). Metabolomic and proteomic analyses revealed that GP binds to carbamoyl phosphate synthetase 1 (CPS1), triggering urea cycle dysfunction marked by citrulline depletion and significant downregulation of its downstream product fumarate. In vitro experiments confirmed that disrupting CPS1 impairs the urea cycle and induces oxidative stress, while CPS1 overexpression or exogenous fumarate supplementation significantly reversed oxidative stress. Clinical sample analysis revealed similar urea cycle metabolic disturbances-characterized by citrulline depletion and reduced fumarate levels−in plasma from patients with liver injury due to inappropriate use of Gardeniae Fructus-containing formulas, confirming clinical relevance. This study reveals a novel hepatotoxic mechanism whereby GP targets CPS1 to disrupt the urea cycle, reducing fumarate production and impairing nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated antioxidant defense, thereby inducing oxidative stress. These findings provide a scientific basis for improving the safety evaluation and rational clinical use of Gardeniae Fructus-containing preparations.
- Gardeniae Fructus,
- Genipin,
- Carbamoyl phosphate synthetase 1,
- Urea cycle

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References(50)

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