Unraveling pyrrolizidine alkaloid-induced liver damage with an integrative spatial lipidomics framework

Yilin Chen; Jie Xu; Thomas Ka-Yam Lam; Yanqiao Xie; Jianing Wang; Aizhen Xiong; Zhengtao Wang; Zongwei Cai; Linnan Li; Li Yang

doi:10.1016/j.jpha.2025.101340

Volume 16 Issue 1

Jan. 2026

Turn off MathJax

Article Contents

Article Navigation > Journal of Pharmaceutical Analysis > 2026 > 16(1): 101340

Yilin Chen, Jie Xu, Thomas Ka-Yam Lam, Yanqiao Xie, Jianing Wang, Aizhen Xiong, Zhengtao Wang, Zongwei Cai, Linnan Li, Li Yang. Unraveling pyrrolizidine alkaloid-induced liver damage with an integrative spatial lipidomics framework[J]. Journal of Pharmaceutical Analysis, 2026, 16(1): 101340. doi: 10.1016/j.jpha.2025.101340

Citation:

Yilin Chen, Jie Xu, Thomas Ka-Yam Lam, Yanqiao Xie, Jianing Wang, Aizhen Xiong, Zhengtao Wang, Zongwei Cai, Linnan Li, Li Yang. Unraveling pyrrolizidine alkaloid-induced liver damage with an integrative spatial lipidomics framework[J]. Journal of Pharmaceutical Analysis, 2026, 16(1): 101340. doi: 10.1016/j.jpha.2025.101340

Citation:

Yilin Chen, Jie Xu, Thomas Ka-Yam Lam, Yanqiao Xie, Jianing Wang, Aizhen Xiong, Zhengtao Wang, Zongwei Cai, Linnan Li, Li Yang. Unraveling pyrrolizidine alkaloid-induced liver damage with an integrative spatial lipidomics framework[J]. Journal of Pharmaceutical Analysis, 2026, 16(1): 101340. doi: 10.1016/j.jpha.2025.101340

PDF( 5255 KB)

Unraveling pyrrolizidine alkaloid-induced liver damage with an integrative spatial lipidomics framework

doi: 10.1016/j.jpha.2025.101340

a. State Key Laboratory of Discovery and Utilization of Functional Components in Traditional Chinese Medicine, The MOE Key Laboratory of Standardization of Chinese Medicines, Shanghai Key Laboratory of Compound Chinese Medicines, SATCM Key Laboratory of New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China;
b. Department of Pharmacy, Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai, 200137, China;
c. State Key Laboratory of Environmental and Biological Analysis, Department of Chemistry, Hong Kong Baptist University, Kowloon 999077 Hong Kong SAR, China

Funds:

This work is financially supported by the National Natural Science Foundation of China (Grant Nos.: 82274223, 82474199, and 82404982) and National Key R&

D Program of China (Grant No.: 2024YFC3506600)\. We would like to express our gratitude to the laboratory staff, Dr. Hongyuan Hao, Dr. Mengxuan Chen and Ms. Fangli Zhang, from Shimadzu (China) Co., Ltd. for their technical assistance with the MALDI-MSI experiments.

Received Date: Feb. 14, 2025
Accepted Date: May 08, 2025
Rev Recd Date: Apr. 24, 2025
Publish Date: May 12, 2025

Abstract

Abstract

Pyrrolizidine alkaloids (PAs), a class of secondary metabolites widely distributed in plants and the accidental ingestion or improper use of foods and herbs containing PAs, can lead to irreversible liver damage. Considering that the toxic mechanism of PAs is closely associated with metabolism, the hepatotoxicity was analyzed from the perspective of lipid metabolism. An integrated analytical approach was employed, combining mass spectrometry imaging (MSI) with liquid chromatography-mass spectrometry (LC-MS), to comprehensively investigate the spatial and temporal dynamics of lipid metabolites during PA exposure. The final lipidomics results combined with RNA sequencing showed that time-dependent changes in metabolite levels after the administration of PAs, involving the pathways of fatty acids, glycerophospholipids, glycerolipids and sphingolipids. Among them, phosphatidylcholines (PC), phosphatidylethanolamines (PE), phosphatidylinositols (PI) and sphingomyelins (SM) were downregulated to varying degrees within 0–24 h, while phosphatidylglycerol (PG), ceramides (Cer), diacylglycerols (DG) and triacylglycerols (TG) were upregulated. Notably, certain lipids exhibited distinct spatial distributions; for example, elevated levels of TG (56:13) were localized near the hepatic portal vein. Subsequently, the changes of lipid subclasses recovered within 24–48 h. Transcriptome RNA sequencing was used to enrich for key pathway-related differential genes Pemt, Gpat, etc. to explain the regulation of the hepatotoxic lipid pathway. The integration of MSI with LC-MS spectroscopy of endogenous metabolites provided intuitive insights into the alterations and spatial distribution of lipid metabolism in mice. Consequently, this study may enhance specific assessments and facilitate early diagnosis of acute toxicity associated with PAs.
- Mass spectrometry imaging,
- DESI,
- Lipidomics,
- Pyrrolizidine alkaloids

FullText(HTML)

References(46)

References

[1]	T. Kopp, M. Abdel-Tawab, B. Mizaikoff, Extracting and analyzing pyrrolizidine alkaloids in medicinal plants: A review, Toxins (Basel) 12 (2020), 320.
[2]	Z. Song, W. Lian, Y. He, et al., Targeting erythrocyte-mediated hypoxia to alleviate lung injury induced by pyrrolizidine alkaloids, Arch. Toxicol. 97 (2023) 819-829.
[3]	Y. He, L. Zhu, J. Ma, et al., Metabolism-mediated cytotoxicity and genotoxicity of pyrrolizidine alkaloids, Arch. Toxicol. 95 (2021) 1917-1942.
[4]	L. Chen, P.P.J. Mulder, A. Peijnenburg, et al., Risk assessment of intake of pyrrolizidine alkaloids from herbal teas and medicines following realistic exposure scenarios, Food Chem. Toxicol. 130 (2019) 142-153.
[5]	P.P.J. Mulder, P. Lopez, M. Castellari, et al., Occurrence of pyrrolizidine alkaloids in animal- and plant-derived food: results of a survey across Europe, Food Addit Contam Part A Chem Anal Control Expo Risk Assess 35 (2018) 118-133.
[6]	L. Zhu, C. Zhang, D. Li, et al., Tu-San-qi (Gynura japonica): The culprit behind pyrrolizidine alkaloid-induced liver injury in China, Acta Pharmacol. Sin. 42 (2021) 1212-1222.
[7]	P.A. Patel-Rodrigues, L. Cundra, D. Alhaqqan, et al., Herbal- and dietary-supplement-induced liver injury: A review of the recent literature, Livers 4 (2024) 94-118.
[8]	S. Soylu, A. CevIk, N. TImurkaan, Pathological and immunohistochemical changes in the liver of monocrotaline-treated rats, Turk. J. Vet. Anim. Sci. 46 (2022) 795-802.
[9]	A. Xiong, Y. Shao, L. Fang, et al., Comparative analysis of toxic components in different medicinal parts of Gynura japonica and its toxicity assessment on mice, Phytomedicine 54 (2019) 77-88.
[10]	W. Zhang, L. Liu, M. Zhang, et al., Validation of the Nanjing criteria for diagnosing pyrrolizidine alkaloids-induced hepatic sinusoidal obstruction syndrome, J. Clin. Transl. Hepatol. 9 (2021) 345-352.
[11]	J. Xu, A. Xiong, X. Wang, et al., Hyperoside attenuates pyrrolizidine alkaloids-induced liver injury by ameliorating TFEB-mediated mitochondrial dysfunction, Arch. Pharm. Res. 46 (2023) 694-712.
[12]	Y. Chen, W. Wang, X. Jia, et al., Firm evidence for the detoxification of senecionine-induced hepatotoxicity via N-glucuronidation in UGT1A4-humanized transgenic mice, Food Chem. Toxicol. 165 (2022), 113185.
[13]	Y. Pan, J. Ma, H. Zhao, et al., Hepatotoxicity screening and ranking of structurally different pyrrolizidine alkaloids in zebrafish, Food Chem. Toxicol. 178 (2023), 113903.
[14]	J. Ma, J. Ruan, X. Chen, et al., Pyrrole-hemoglobin adducts, a more feasible potential biomarker of pyrrolizidine alkaloid exposure, Chem. Res. Toxicol. 32 (2019) 1027-1039.
[15]	Y. Chen, L. Li, J. Xu, et al., Mass spectrometric analysis strategies for pyrrolizidine alkaloids, Food Chem. 445 (2024), 138748.
[16]	Y. Li, Y. Tian, Q. Wang, et al., Serum metabolomics strategy for investigating the hepatotoxicity induced by different exposure times and doses of Gynura segetum (Lour.) Merr. in rats based on GC-MS, RSC Adv. 13 (2023) 2635-2648.
[17]	Y. Dai, Q. Guo, K. Xu, et al., Lactational retrorsine exposure changes maternal milk components and disturbs metabolism homeostasis of offspring rats, Sci. Total Environ. 894 (2023), 164929.
[18]	A. Xiong, L. Lu, K. Jiang, et al., Functional metabolomics characterizes the contribution of farnesoid X receptor in pyrrolizidine alkaloid-induced hepatic sinusoidal obstruction syndrome, Arch. Toxicol. 98 (2024) 2557-2576.
[19]	K. Huber, J. Saltzmann, S. Daenicke, Metabolite profiling in the liver, plasma and milk of dairy cows exposed to tansy ragwort (Senecio jacobae) pyrrolizidine alkaloids, Toxins (Basel) 15 (2023), 601.
[20]	A.L. Santos, G. Preta, Lipids in the cell: Organisation regulates function, Cell. Mol. Life Sci. 75 (2018) 1909-1927.
[21]	T. Harayama, H. Riezman, Understanding the diversity of membrane lipid composition, Nat. Rev. Mol. Cell Biol. 19 (2018) 281-296.
[22]	Q. Xiao, M. Xia, W. Tang, et al., The lipid metabolism remodeling: A hurdle in breast cancer therapy, Cancer Lett. 582 (2024), 216512.
[23]	K. Zan, W. Lei, Y. Li, et al., Integrative metabolomics and proteomics detected hepatotoxicity in mice associated with alkaloids from Eupatorium fortunei Turcz, Toxins (Basel) 14 (2022), 765.
[24]	M. Yang, J. Ma, J. Ruan, et al., Intestinal and hepatic biotransformation of pyrrolizidine alkaloid N-oxides to toxic pyrrolizidine alkaloids, Arch. Toxicol. 93 (2019) 2197-2209.
[25]	H. Gao, Y. Lv, W. Zhao, et al., Accurate lamb origin identification and molecular differentiation analysis using rapid evaporative ionization mass spectrometry, Anal. Methods 17 (2025) 1643-1647.
[26]	Q. Xuan, C. Hu, D. Yu, et al., Development of a high coverage pseudotargeted lipidomics method based on ultra-high performance liquid chromatography-mass spectrometry, Anal. Chem. 90 (2018) 7608-7616.
[27]	Y. Zhu, Q. Zang, Z. Luo, et al., An organ-specific metabolite annotation approach for ambient mass spectrometry imaging reveals spatial metabolic alterations of a whole mouse body, Anal. Chem. 94 (2022) 7286-7294.
[28]	C. Zhao, P. Xie, T. Yong, et al., MALDI-MS imaging reveals asymmetric spatial distribution of lipid metabolites from bisphenol S-induced nephrotoxicity, Anal. Chem. 90 (2018) 3196-3204.
[29]	C. Zhao, T. Yong, Y. Zhang, et al., Evaluation of the splenic injury following exposure of mice to bisphenol S: A mass spectrometry-based lipidomics and imaging analysis, Environ. Int. 135 (2020), 105378.
[30]	S.N. Nguyen, J.E. Kyle, S.E. Dautel, et al., Lipid coverage in nanospray desorption electrospray ionization mass spectrometry imaging of mouse lung tissues, Anal. Chem. 91 (2019) 11629-11635.
[31]	I. Kaya, S. Samfors, M. Levin, et al., Multimodal MALDI imaging mass spectrometry reveals spatially correlated lipid and protein changes in mouse heart with acute myocardial infarction, J. Am. Soc. Mass Spectrom. 31 (2020) 2133-2142.
[32]	W. Tang, J. Chen, J. Zhou, et al., Quantitative MALDI imaging of spatial distributions and dynamic changes of tetrandrine in multiple organs of rats, Theranostics 9 (2019) 932-944.
[33]	Y. Chen, Y. Xie, L. Li, et al., Advances in mass spectrometry imaging for toxicological analysis and safety evaluation of pharmaceuticals, Mass Spectrom. Rev. 42 (2023) 2207-2233.
[34]	H. Jiang, H. Gao, J. Li, et al., Integrated spatially resolved metabolomics and network toxicology to investigate the hepatotoxicity mechanisms of component D of Polygonum multiflorum Thunb, J. Ethnopharmacol. 298 (2022), 115630.
[35]	I. Rzagalinski, N. Hainz, C. Meier, et al., MALDI mass spectral imaging of bile acids observed as deprotonated molecules and proton-bound dimers from mouse liver sections, J. Am. Soc. Mass Spectrom. 29 (2018) 711-722.
[36]	R. Matsuyama, Y. Okada, S. Shimma, Metabolite alteration analysis of acetaminophen-induced liver injury using a mass microscope, Anal. Bioanal. Chem. 414 (2022) 3709-3718.
[37]	T. He, K. Lin, L. Xiong, et al., Disorder of phospholipid metabolism in the renal cortex and medulla contributes to acute tubular necrosis in mice after cantharidin exposure using integrative lipidomics and spatial metabolomics, J. Pharm. Anal., (2025), 101210.
[38]	S. Sezgin, R. Hassan, S. Zuhlke, et al., Spatio-temporal visualization of the distribution of acetaminophen as well as its metabolites and adducts in mouse livers by MALDI MSI, Arch. Toxicol. 92 (2018) 2963-2977.
[39]	W. Otkur, Y. Zhang, Y. Li, et al., Spatial multi-omics characterizes GPR35-relevant lipid metabolism signatures across liver zonation in MASLD, Life Metab. 3 (2024), loae021.
[40]	H. Zhang, X. Shao, H. Zhao, et al., Integration of metabolomics and lipidomics reveals metabolic mechanisms of triclosan-induced toxicity in human hepatocytes, Environ. Sci. Technol. 53 (2019) 5406-5415.
[41]	L. Fang, A. Xiong, X. Yang, et al., Mass-spectrometry-directed analysis and purification of pyrrolizidine alkaloid Cis/trans isomers in Gynura japonica, J. Sep. Sci. 37 (2014) 2032-2038.
[42]	D. Hornburg, S. Wu, M. Moqri, et al., Dynamic lipidome alterations associated with human health, disease and ageing, Nat. Metab. 5 (2023) 1578-1594.
[43]	H.J. Yang, K.H. Park, S. Shin, et al., Characterization of heme ions using maldi-tof ms and maldi ft-icr ms, Int. J. Mass Spectrom. 343 (2013) 37-44.
[44]	Y. Zhuge, Y. Liu, W. Xie, et al., Expert consensus on the clinical management of pyrrolizidine alkaloid-induced hepatic sinusoidal obstruction syndrome, J. Gastroenterol. Hepatol. 34 (2019) 634-642.
[45]	J.N. van der Veen, J.P. Kennelly, S. Wan, et al., The critical role of phosphatidylcholine and phosphatidylethanolamine metabolism in health and disease, Biochim. Biophys. Acta Biomembr. 1859 (2017) 1558-1572.
[46]	Y. Miura, E. Gotoh, F. Nara, et al., Hydrolysis of sphingosylphosphocholine by neutral sphingomyelinases, FEBS Lett. 557 (2004) 288-292.