Skip to main content
SpringerLink
Log in

First evidence of pyrrolizidine alkaloid N-oxide-induced hepatic sinusoidal obstruction syndrome in humans

  • Organ Toxicity and Mechanisms
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

Pyrrolizidine alkaloids (PAs) are among the most potent phytotoxins widely distributed in plant species around the world. PA is one of the major causes responsible for the development of hepatic sinusoidal obstruction syndrome (HSOS) and exerts hepatotoxicity via metabolic activation to form the reactive metabolites, which bind with cellular proteins to generate pyrrole-protein adducts, leading to hepatotoxicity. PA N-oxides coexist with their corresponding PAs in plants with varied quantities, sometimes even higher than that of PAs, but the toxicity of PA N-oxides remains unclear. The current study unequivocally identified PA N-oxides as the sole or predominant form of PAs in 18 Gynura segetum herbal samples ingested by patients with liver damage. For the first time, PA N-oxides were recorded to induce HSOS in human. PA N-oxide-induced hepatotoxicity was further confirmed on mice orally dosed of herbal extract containing 170 μmol PA N-oxides/kg/day, with its hepatotoxicity similar to but potency much lower than the corresponding PAs. Furthermore, toxicokinetic study after a single oral dose of senecionine N-oxide (55 μmol/kg) on rats revealed the toxic mechanism that PA N-oxides induced hepatotoxicity via their biotransformation to the corresponding PAs followed by the metabolic activation to form pyrrole-protein adducts. The remarkable differences in toxicokinetic profiles of PAs and PA N-oxides were found and attributed to their significantly different hepatotoxic potency. The findings of PA N-oxide-induced hepatotoxicity in humans and rodents suggested that the contents of both PAs and PA N-oxides present in herbs and foods should be regulated and controlled in use.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Björnsson ES (2015) Drug-induced liver injury: an overview over the most critical compounds. Arch Toxicol 89:327–334

    Article  PubMed  Google Scholar 

  • Bodi D, Ronczka S, Gottschalk C et al (2014) Determination of pyrrolizidine alkaloids in tea, herbal drugs and honey. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 31:1886–1895

    Article  CAS  PubMed  Google Scholar 

  • Cao Y, Colegate SM, Edgar JA (2008) Safety assessment of food and herbal products containing hepatotoxic pyrrolizidine alkaloids: interlaboratory consistency and the importance of N-oxide determination. Phytochem Anal 19:526–533

    Article  CAS  PubMed  Google Scholar 

  • Chou MW, Wang YP, Yan J et al (2003) Riddelliine N-oxide is a phytochemical and mammalian metabolite with genotoxic activity that is comparable to the parent pyrrolizidine alkaloid riddelliine. Toxicol Lett 145:239–247

    Article  CAS  PubMed  Google Scholar 

  • Chu PS, Lamé MW, Segall HJ (1993) In vivo metabolism of retrorsine and retrorsine-N-oxide. Arch Toxicol 67:39–43

    Article  CAS  PubMed  Google Scholar 

  • Dai N, Yu Y-C, Ren T-H et al (2007) Gynura root induces hepatic veno-occlusive disease: a case report and review of the literature. World J Gastroenterol 13:1628–1631

    Article  PubMed  PubMed Central  Google Scholar 

  • Dick AT, Dann A, Bull L, Culvenok C (1963) Vitamin B12 and the detoxification of hepatotoxic pyrrolizidine alkaloids in rumen liquor. Nature 197:207–208

    Article  CAS  Google Scholar 

  • Edgar JA, Molyneux RJ, Colegate SM (2015) Pyrrolizidine alkaloids: potential role in the etiology of cancers, pulmonary hypertension, congenital anomalies, and liver disease. Chem Res Toxicol 28:4–20

    Article  CAS  PubMed  Google Scholar 

  • Fan CQ, Crawford JM (2014) Sinusoidal obstruction syndrome (hepatic veno-occlusive disease). J Clin Exp Hepatol 4:332–346

    Article  PubMed  PubMed Central  Google Scholar 

  • Field RA, Stegelmeier BL, Colegate SM, Brown AW, Green BT (2015) An in vitro comparison of the cytotoxic potential of selected dehydropyrrolizidine alkaloids and some N-oxides. Toxicon 97:36–45

    Article  CAS  PubMed  Google Scholar 

  • Fu PP, Yang Y-C, Xia Q, Chou MW, Cui YY, Lin G (2002) Pyrrolizidine alkaloids—tumorigenic components in Chinese herbal medicines and dietary supplements. J Food Drug Anal 10:198–211

    CAS  Google Scholar 

  • Fu PP, Xia Q, Lin G, Chou MW (2004) Pyrrolizidine alkaloids–genotoxicity, metabolism enzymes, metabolic activation, and mechanisms. Drug Metab Rev 36:1–55

    Article  CAS  PubMed  Google Scholar 

  • Gao H, Li N, Wang JY, Zhang SC, Lin G (2012) Definitive diagnosis of hepatic sinusoidal obstruction syndrome induced by pyrrolizidine alkaloids. J Dig Dis 13:33–39

    Article  PubMed  Google Scholar 

  • Gao H, Ruan JQ, Chen J et al (2015) Blood pyrrole-protein adducts as a diagnostic and prognostic index in pyrrolizidine alkaloid-hepatic sinusoidal obstruction syndrome. Drug Des Devel Ther 9:4861–4868

    Article  PubMed  PubMed Central  Google Scholar 

  • Helmy A (2006) Review article: updates in the pathogenesis and therapy of hepatic sinusoidal obstruction syndrome. Aliment Pharmacol Ther 23:11–25

    Article  CAS  PubMed  Google Scholar 

  • Kakar F, Akbarian Z, Leslie T et al (2010) An outbreak of hepatic veno-occlusive disease in Western afghanistan associated with exposure to wheat flour contaminated with pyrrolizidine alkaloids. J Toxicol 2010:313280

    Article  PubMed  PubMed Central  Google Scholar 

  • Kempf M, Reinhard A, Beuerle T (2010) Pyrrolizidine alkaloids (PAs) in honey and pollen-legal regulation of PA levels in food and animal feed required. Mol Nutr Food Res 54:158–168

    Article  CAS  PubMed  Google Scholar 

  • Li SL, Lin G, Fu PP et al (2008) Identification of five hepatotoxic pyrrolizidine alkaloids in a commonly used traditional Chinese medicinal herb, Herba Senecionis scandentis (Qianliguang). Rapid Commun Mass Spectrom 22:591–602

    Article  CAS  PubMed  Google Scholar 

  • Lin G, Zhou KY, Zhao XG et al (1998) Determination of hepatotoxic pyrrolizidine alkaloids by on-line high performance liquid chromatography mass spectrometry with an electrospray interface. Rapid Commun Mass Spectrom 12:1445–1456

    Article  CAS  PubMed  Google Scholar 

  • Lin G, Rose P, Chatson KB, Hawes EM, Zhao XG, Wang ZT (2000) Characterization of two structural forms of otonecine-type pyrrolizidine alkaloids from Ligularia hodgsonii by NMR spectroscopy. J Nat Prod 63:857–860

    Article  CAS  PubMed  Google Scholar 

  • Lin G, Li SL, Li M et al (2009) Qianliguang (Senecio scandens) safety dilemma: dose is the key? Planta Med 75:1107–1111

    Article  CAS  PubMed  Google Scholar 

  • Lin G, Wang JY, Li N et al (2011) Hepatic sinusoidal obstruction syndrome associated with consumption of Gynura segetum. J Hepatol 54:666–673

    Article  PubMed  Google Scholar 

  • Mattocks AR (1971a) The occurrence and analysis of pyrrolizidine alkaloid N-oxides. Xenobiotica 1(4):451–453

    Article  CAS  PubMed  Google Scholar 

  • Mattocks AR (1971b) Hepatotoxic effects due to pyrrolizidine alkaloid N-oxides. Xenobiotica 1:563–565

    Article  CAS  PubMed  Google Scholar 

  • Mattocks AR (1986) Chemistry and toxicology of pyrrolizidine alkaloids. Academic Press, London

    Google Scholar 

  • Mattocks AR, Bird I (1983) Pyrrolic and N-oxide metabolites formed from pyrrolizidine alkaloids by hepatic microsomes in vitro: relevance to in vivo hepatotoxicity. Chem Biol Interact 43:209–222

    Article  CAS  PubMed  Google Scholar 

  • Molyneux R, Gardner D, Colegate S, Edgar J (2011) Pyrrolizidine alkaloid toxicity in livestock: a paradigm for human poisoning? Food Addit Contam Part A 28:293–307

    Article  CAS  Google Scholar 

  • Navarro VJ, Barnhart H, Bonkovsky HL et al (2014) Liver injury from herbals and dietary supplements in the US Drug-Induced Liver Injury Network. Hepatology 60:1399–1408

    Article  PubMed  PubMed Central  Google Scholar 

  • Robinson O, Want E, Coen M et al (2013) Hirmi valley liver disease: a disease associated with exposure to pyrrolizidine alkaloids and DDT. J Hepatol 60:96–102

    Article  PubMed  Google Scholar 

  • Ruan J, Li N, Xia Q, Fu PP, Peng S, Ye Y, Lin G (2012) Characteristic ion clusters as determinants for the identification of pyrrolizidine alkaloid N-oxides in pyrrolizidine alkaloid-containing natural products using HPLC–MS analysis. J Mass Spectrom 47:331–337

    Article  CAS  PubMed  Google Scholar 

  • Ruan J, Yang M, Fu P, Ye Y, Lin G (2014) Metabolic activation of pyrrolizidine alkaloids: insights into the structural and enzymatic basis. Chem Res Toxicol 27:1030–1039

    Article  CAS  PubMed  Google Scholar 

  • Ruan J, Gao H, Li N et al (2015) Blood pyrrole-protein adducts—a biomarker of pyrrolizidine alkaloid-induced liver injury in humans. J Environ Sci Health C 33(4):404–421

    Article  CAS  Google Scholar 

  • Schneider J, Tsegaye Y, W/Tensae M et al (2012) Veno-occlusive liver disease: a case report. Ethiop Med J 50(Suppl 2):47–51

    PubMed  Google Scholar 

  • Smith LW, Culvenor CC (1981) Plant sources of hepatotoxic pyrrolizidine alkaloids. J Nat Prod 44:129–152

    Article  CAS  PubMed  Google Scholar 

  • Song Y, Fan YH (2011) Clinical features of hepatic veno-occlusive disease induced by gynura root: analysis of 102 cases. Lin Chuang Gan Dan Bing Za Zhi 5:015

    Google Scholar 

  • Stickel F, Shouval D (2015) Hepatotoxicity of herbal and dietary supplements: an update. Arch Toxicol 89:851–865

    Article  CAS  PubMed  Google Scholar 

  • US Food and Drug Administration (2005) Guidance for industry: estimating the maximum safe starting dose in initial clinical trials for therapeutics in adult healthy volunteers. Center for Drug Evaluation and Research (CDER)

  • Wang YP, Yan J, Fu PP, Chou MW (2005) Human liver microsomal reduction of pyrrolizidine alkaloid N-oxides to form the corresponding carcinogenic parent alkaloid. Toxicol Lett 155:411–420

    Article  CAS  PubMed  Google Scholar 

  • Wang X, Qi X, Guo X (2015) Tusanqi-related sinusoidal obstruction syndrome in China: a systematic review of the literatures. Medicine 94:e942

    Article  PubMed  PubMed Central  Google Scholar 

  • Williams D, Reed R, Kedzierski B, Dannan G, Guengerich F, Buhler D (1989) Bioactivation and detoxication of the pyrrolizidine alkaloid senecionine by cytochrome P-450 enzymes in rat liver. Drug Metab Dispos 17(4):387–392

    CAS  PubMed  Google Scholar 

  • Williams MT, Warnock BJ, Betz JM et al (2011) Detection of high levels of pyrrolizidine-N-oxides in the endangered plant Cryptantha crassipes (Terlingua Creek Cat’s-eye) using HPLC-ESI-MS. Phytochem Anal 22:532–540

    Article  CAS  PubMed  Google Scholar 

  • Willmot F, Robertson G (1920) Senecio disease, or cirrhosis of the liver due to Senecio poisoning. Lancet 196(5069):848–849

    Article  Google Scholar 

  • Yang M, Ruan J, Fu PP, Lin G (2016) Cytotoxicity of pyrrolizidine alkaloid in human hepatic parenchymal and sinusoidal endothelial cells: firm evidence for the reactive metabolites mediated pyrrolizidine alkaloid-induced hepatotoxicity. Chem Biol Interact 243:119–126

    Article  CAS  PubMed  Google Scholar 

  • Zhu L, Ruan J, Li N, Fu PP, Ye Y, Lin G (2016) A novel ultra-performance liquid chromatography hyphenated with quadrupole time of flight mass spectrometry method for rapid estimation of total toxic retronecine-type of pyrrolizidine alkaloids in herbs without requiring corresponding standards. Food Chem 194:1320–1328

    Article  CAS  PubMed  Google Scholar 

  • Zhu L, Xue J, Xia Q, Fu PP, Lin G (2017) The long persistence of pyrrolizidine alkaloid-derived DNA adducts in vivo: kinetic study following single and multiple exposures in male ICR mice. Arch Toxicol 91:949–965

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The present study was supported by Research Grant Council of Hong Kong (GRF Grants Nos. 471013 and 14110714), One-off Funding for Joint Lab/Research Collaboration (Project Code: 3132968) by CUHK, and CUHK School of Biomedical Sciences–Seed Fund for Joint Establishments. This article is not an official US Food and Drug Administration (FDA) guidance or policy statement. No official support or endorsement by the US FDA is intended or should be inferred. Funding was provided by Chinese University of Hong Kong (Grants Nos. 4054302 and 3132968).

Author information

Author notes

  1. Mengbi Yang and Jianqing Ruan contributed equally to this work and shared the first authorship.

Authors and Affiliations

  1. School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, SAR, Hong Kong

    Mengbi Yang, Jianqing Ruan, Na Li, Jiang Ma, Junyi Xue & Ge Lin

  2. Division of Gastroenterology, Zhongshan Hospital, Fudan University, Shanghai, People’s Republic of China

    Hong Gao & Jiyao Wang

  3. Joint Research Laboratory for Promoting Globalization of Traditional Chinese Medicines Between Shanghai Institute of Materia Medica, Chinese Academy of Sciences and The Chinese University of Hong Kong, Hong Kong, People’s Republic of China

    Mengbi Yang, Jianqing Ruan, Jiang Ma, Junyi Xue, Yang Ye & Ge Lin

  4. State Key Laboratory of Drug Research and Natural Products Chemistry Department, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, People’s Republic of China

    Yang Ye

  5. National Center for Toxicological Research, Jefferson, AR, USA

    Peter Pi-Cheng Fu

Authors
  1. Mengbi Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jianqing Ruan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hong Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Na Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jiang Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Junyi Xue
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yang Ye
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Peter Pi-Cheng Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jiyao Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Ge Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ge Lin.

Ethics declarations

All procedures performed in studies involving human participants were approval by Ethics Committee of Fudan University, Shanghai, China. All procedures performed in studies involving animals were approval by Animal Experimental Ethics Committee, The Chinese University of Hong Kong, Hong Kong, China.

Conflict of interest

The authors declare no competing financial interest.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PPTX 180 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yang, M., Ruan, J., Gao, H. et al. First evidence of pyrrolizidine alkaloid N-oxide-induced hepatic sinusoidal obstruction syndrome in humans. Arch Toxicol 91, 3913–3925 (2017). https://doi.org/10.1007/s00204-017-2013-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-017-2013-y

Keywords

Search

Navigation