Abstract
Pyrrolizidine alkaloids (PAs) are phytotoxins widely present in various natural products and foodstuffs. The present study aims to investigate the effects of fasting on PA-induced hepatotoxicity and the underlying biochemical mechanisms. The results of hepatotoxic study showed that 15-h overnight fasting significantly exacerbated the hepatotoxicity of retrorsine (RTS, a representative toxic PA) in fasted rats compared to fed rats, as indicated by remarkably elevated plasma ALT and bilirubin levels and obvious liver histological changes. Further toxicokinetic studies revealed that fasting significantly enhanced cytochromes P450 enzymes (CYPs)-mediated metabolic activation of RTS leading to increased formation of pyrrole-protein adducts and thus decreased the in vivo exposure and excretion of both parent RTS and its N-oxide metabolite. Metabolic studies demonstrated that fasting induced enzyme activities of CYP1A2, CYP2B6 and CYP2E1 that participated in catalyzing RTS to its reactive pyrrolic metabolites. Moreover, fasting also dramatically decreased hepatic glutathione (GSH) content, which restricted the detoxification of GSH by neutralizing the reactive pyrrolic metabolite of RTS, further contributing to the enhanced hepatotoxicity. The present findings may have an impact on future PA toxicity tests with different dietary styles and/or risk assessment of metabolite-mediated toxins by considering the profound effects of fasting.
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Abbreviations
- CYPs:
-
Cytochrome P450 enzymes
- Dehydro-PAs:
-
Dehydro-pyrrolizidine alkaloids
- GSH:
-
Glutathione
- H&E staining:
-
Hematoxylin–eosin staining
- MRM:
-
Multiple reaction monitoring
- NADPH:
-
Reduced nicotinamide adenine dinucleotide phosphate
- PAs:
-
Pyrrolizidine alkaloids
- Pyrrole-GSH:
-
7,9-Diglutathione-(±)-6,7-dihydro-7-hydroxy-1-hydroxymethyl-5H-pyrrolizine
- RTS:
-
Retrorsine
- UHPLC-MS/MS:
-
Ultrahigh pressure liquid chromatography-triple quadrupole/mass spectrometry
References
Baillie TA, Rettie AE (2011) Role of biotransformation in drug-induced toxicity: influence of intra- and inter-species differences in drug metabolism. Drug Metab Pharmacokinet 26(1):15–29. https://doi.org/10.2133/dmpk.dmpk-10-rv-089
Brown BL, Allis JW, Simmons JE, House DE (1995) Fasting for less than 24 h induces cytochrome P450 2E1 and 2B1/2 activities in rats. Toxicol Lett 81(1):39–44. https://doi.org/10.1016/0378-4274(95)03407-2
Chauvin P, Dillon JC, Moren A (1994) An outbreak of Heliotrope food poisoning, Tadjikistan, November 1992-March 1993. Sante 4(4):263–268
Chojkier M (2003) Hepatic sinusoidal-obstruction syndrome: toxicity of pyrrolizidine alkaloids. J Hepatol 39(3):437–446. https://doi.org/10.1016/S0168-8278(03)00231-9
DeLeve LD, Wang XD, Kuhlenkamp JF, Kaplowitz N (1996) Toxicity of azathioprine and monocrotaline in murine sinusoidal endothelial cells and hepatocytes: the role of glutathione and relevance to hepatic venoocclusive disease. Hepatology 23(3):589–599
DeLeve LD, Ito Y, Bethea NW, McCuskey MK, Wang XD, McCuskey RS (2003) Embolization by sinusoidal lining cells obstructs the microcirculation in rat sinusoidal obstruction syndrome. Am J Physiol-Gastr L 284(6):G1045–G1052. https://doi.org/10.1152/ajpgi.00526.2002
Deleve LD, Kaplowitz N (1991) Glutathione metabolism and its role in hepatotoxicity. Pharmacol Therapeut 52(3):287–305. https://doi.org/10.1016/0163-7258(91)90029-L
Dusemund B, Nowak N, Sommerfeld C, Lindtner O, Schafer B, Lampen A (2018) Risk assessment of pyrrolizidine alkaloids in food of plant and animal origin. Food Chem Toxicol 115:63–72. https://doi.org/10.1016/j.fct.2018.03.005
Ebmeyer J, Rasinger JD, Hengstler JG et al (2020) Hepatotoxic pyrrolizidine alkaloids induce DNA damage response in rat liver in a 28-day feeding study. Arch Toxicol. https://doi.org/10.1007/s00204-020-02779-2
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(1):4–20. https://doi.org/10.1021/tx500403t
Fu PP (2017) Pyrrolizidine alkaloids: metabolic activation pathways leading to liver tumor initiation. Chem Res Toxicol 30(1):81–93. https://doi.org/10.1021/acs.chemrestox.6b00297
Fu PP, Xia QS, Lin G, Chou MW (2004) Pyrrolizidine alkaloid—genotoxicity, metabolism enzymes, metabolic activation, and mechanisms. Drug Metab Rev 36(1):1–55. https://doi.org/10.1081/Dmr-120028426
He Y, Zhu L, Ma J, Lin G (2021) Metabolism-mediated cytotoxicity and genotoxicity of pyrrolizidine alkaloids. Arch Toxicol 95(6):1917–1942. https://doi.org/10.1007/s00204-021-03060-w
Hessel-Pras S, Braeuning A, Guenther G et al (2020) The pyrrolizidine alkaloid senecionine induces CYP-dependent destruction of sinusoidal endothelial cells and cholestasis in mice. Arch Toxicol 94(1):219–229. https://doi.org/10.1007/s00204-019-02582-8
Hinson JA, Pumford NR, Nelson SD (1994) The role of metabolic activation in drug toxicity. Drug Metab Rev 26(1–2):395–412. https://doi.org/10.3109/03602539409029805
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. https://doi.org/10.1155/2010/313280
Lammers LA, Achterbergh R, van Schaik RHN, Romijn JA, Mathot RAA (2017) Effect of short-term fasting on systemic cytochrome P450-mediated drug metabolism in healthy subjects: a randomized, controlled, crossover study using a cocktail approach. Clin Pharmacokinet 56(10):1231–1244. https://doi.org/10.1007/s40262-017-0515-7
Lin G, Cui YY, Hawes EM (2000) Characterization of rat liver microsomal metabolites of clivorine, an hepatotoxic otonecine-type pyrrolizidine alkaloid. Drug Metab Dispos 28(12):1475–1483
Lin G, Wang JY, Li N et al (2011) Hepatic sinusoidal obstruction syndrome associated with consumption of Gynura segetum. J Hepatol 54(4):666–673. https://doi.org/10.1016/j.jhep.2010.07.031
Longo VD, Panda S (2016) Fasting, circadian rhythms, and time-restricted feeding in healthy lifespan. Cell Metab 23(6):1048–1059. https://doi.org/10.1016/j.cmet.2016.06.001
Lu Y, Ma J, Lin G (2019) Development of a two-layer transwell co-culture model for the in vitro investigation of pyrrolizidine alkaloid-induced hepatic sinusoidal damage. Food Chem Toxicol 129:391–398. https://doi.org/10.1016/j.fct.2019.04.057
Ma L, Zhao H, Xia Q, Cai L, Fu PP (2015) Synthesis and phototoxicity of isomeric 7,9-diglutathione pyrrole adducts: formation of reactive oxygen species and induction of lipid peroxidation. J Food Drug Anal 23(3):577–586. https://doi.org/10.1016/j.jfda.2015.06.001
Ma J, Xia Q, Fu PP, Lin G (2018) Pyrrole-protein adducts—a biomarker of pyrrolizidine alkaloid-induced hepatotoxicity. J Food Drug Anal 26(3):965–972. https://doi.org/10.1016/j.jfda.2018.05.005
Ma J, Ruan J, Chen X et al (2019) Pyrrole-hemoglobin adducts, a more feasible potential biomarker of pyrrolizidine alkaloid exposure. Chem Res Toxicol 32(6):1027–1039. https://doi.org/10.1021/acs.chemrestox.8b00369
Ma J, Zhang W, He Y et al (2021) Clinical application of pyrrole-hemoglobin adducts as a biomarker of pyrrolizidine alkaloid exposure in humans. Arch Toxicol 95(2):759–765. https://doi.org/10.1007/s00204-020-02947-4
Merz KH, Schrenk D (2016) Interim relative potency factors for the toxicological risk assessment of pyrrolizidine alkaloids in food and herbal medicines. Toxicol Lett 263:44–57. https://doi.org/10.1016/j.toxlet.2016.05.002
Moore RA, Derry S, Wiffen PJ, Straube S (2015) Effects of food on pharmacokinetics of immediate release oral formulations of aspirin, dipyrone, paracetamol and NSAIDs—a systematic review. Br J Clin Pharmacol 80(3):381–388. https://doi.org/10.1111/bcp.12628
Patterson RE, Sears DD (2017) Metabolic effects of intermittent fasting. Annu Rev Nutr 37:371–393. https://doi.org/10.1146/annurev-nutr-071816-064634
Pessayre D, Dolder A, Artigou JY et al (1979) Effect of fasting on metabolite-mediated hepatotoxicity in the rat. Gastroenterology 77(2):264–271
Pizzorno J (2014) Glutathione! Integr Med (encinitas) 13(1):8–12
Price VF, Miller MG, Jollow DJ (1987) Mechanisms of fasting-induced potentiation of acetaminophen hepatotoxicity in the rat. Biochem Pharmacol 36(4):427–433
Rasmussen MK, Bertholdt L, Gudiksen A, Pilegaard H, Knudsen JG (2018) Impact of fasting followed by short-term exposure to interleukin-6 on cytochrome P450 mRNA in mice. Toxicol Lett 282:93–99. https://doi.org/10.1016/j.toxlet.2017.10.011
Robinson O, Want E, Coen M et al (2014) Hirmi Valley liver disease: a disease associated with exposure to pyrrolizidine alkaloids and DDT. J Hepatol 60(1):96–102. https://doi.org/10.1016/j.jhep.2013.07.039
Ruan J, Liao C, Ye Y, Lin G (2014a) Lack of metabolic activation and predominant formation of an excreted metabolite of nontoxic platynecine-type pyrrolizidine alkaloids. Chem Res Toxicol 27(1):7–16. https://doi.org/10.1021/tx4004159
Ruan J, Yang M, Fu PP, Ye Y, Lin G (2014b) Metabolic activation of pyrrolizidine alkaloids: insights into the structural and enzymatic basis. Chem Res Toxicol 27(6):1030–1039. https://doi.org/10.1021/tx500071q
Smith PK, Krohn RI, Hermanson GT et al (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150(1):76–85. https://doi.org/10.1016/0003-2697(85)90442-7
Szkudelski T, Okulicz M, Bialik I, Szkudelska K (2004) The influence of fasting on liver sulfhydryl groups, glutathione peroxidase and glutathione-S-transferase activities in the rat. J Physiol Biochem 60(1):1–6. https://doi.org/10.1007/BF03168215
Tateishi N, Higashi T, Shinya S, Naruse A, Sakamoto Y (1974) Studies on the regulation of glutathione level in rat liver. J Biochem 75(1):93–103. https://doi.org/10.1093/oxfordjournals.jbchem.a130387
Turpeinen M, Uusitalo J, Jalonen J, Pelkonen O (2005) Multiple P450 substrates in a single run: rapid and comprehensive in vitro interaction assay. Eur J Pharm Sci 24(1):123–132. https://doi.org/10.1016/j.ejps.2004.10.006
Walker RM, Massey TE, McElligott TF, Racz WJ (1982) Acetaminophen toxicity in fed and fasted mice. Can J Physiol Pharmacol 60(3):399–404
Wang X, Qi X, Guo X (2015) Tusanqi-related sinusoidal obstruction syndrome in China: a systematic review of the literatures. Med (baltim) 94(23):e942. https://doi.org/10.1097/MD.0000000000000942
Whitcomb DC, Block GD (1994) Association of acetaminophen hepatotoxicity with fasting and ethanol use. JAMA 272(23):1845–1850
Willmot FC, Robertson GW (1920) Senecio disease, or cirrhosis of the liver due to senecio poisoning. Lancet 2:848–849
Yan CC, Huxtable RJ (1995) Relationship between glutathione concentration and metabolism of the pyrrolizidine alkaloid, monocrotaline, in the isolated, perfused liver. Toxicol Appl Pharm 130(1):132–139. https://doi.org/10.1006/taap.1995.1017
Yang MB, Ruan JQ, 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. https://doi.org/10.1016/j.cbi.2015.09.011
Yang M, Ma J, Ruan J, Ye Y, Fu PP, Lin G (2019) Intestinal and hepatic biotransformation of pyrrolizidine alkaloid N-oxides to toxic pyrrolizidine alkaloids. Arch Toxicol 93(8):2197–2209. https://doi.org/10.1007/s00204-019-02499-2
Zhou S, Chan E, Duan W, Huang M, Chen YZ (2005) Drug bioactivation, covalent binding to target proteins and toxicity relevance. Drug Metab Rev 37(1):41–213. https://doi.org/10.1081/dmr-200028812
Zhu L, Zhang CY, Li DP et al (2021) Tu-San-Qi (Gynura japonica): the culprit behind pyrrolizidine alkaloid-induced liver injury in China. Acta Pharmacol Sin 42(8):1212–1222. https://doi.org/10.1038/s41401-020-00553-9
Zhuge YZ, Wang Y, Zhang F et al (2018) Clinical characteristics and treatment of pyrrolizidine alkaloid-related hepatic vein occlusive disease. Liver Int 38(10):1867–1874. https://doi.org/10.1111/liv.13684
Acknowledgements
This work was supported by Research Grants Council of Hong Kong (GRF grant: 14106318) and The Chinese University of Hong Kong (Direct Grant: 4054428).
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Ma, J., Zhang, C., He, Y. et al. Fasting augments pyrrolizidine alkaloid-induced hepatotoxicity. Arch Toxicol 96, 639–651 (2022). https://doi.org/10.1007/s00204-021-03193-y
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DOI: https://doi.org/10.1007/s00204-021-03193-y