The pyrrolizidine alkaloid senecionine induces CYP-dependent destruction of sinusoidal endothelial cells and cholestasis in mice

Stefanie Hessel-Pras (Lead / Corresponding author), Albert Braeuning, Georgia Guenther, Alshaimaa Adawy, Anne-Margarethe Enge, Johanna Ebmeyer, Colin J. Henderson, Jan G. Hengstler, Alfonso Lampen, Raymond Reif

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Pyrrolizidine alkaloids (PAs) are widely occurring phytotoxins which can induce severe liver damage in humans and other mammalian species by mechanisms that are not fully understood. Therefore, we investigated the development of PA hepatotoxicity in vivo, using an acutely toxic dose of the PA senecionine in mice, in combination with intravital two-photon microscopy, histology, clinical chemistry, and in vitro experiments with primary mouse hepatocytes and liver sinusoidal endothelial cells (LSECs). We observed pericentral LSEC necrosis together with elevated sinusoidal marker proteins in the serum of senecionine-treated mice and increased sinusoidal platelet aggregation in the damaged tissue regions. In vitro experiments showed no cytotoxicity to freshly isolated LSECs up to 500 µM senecionine. However, metabolic activation of senecionine by preincubation with primary mouse hepatocytes increased the cytotoxicity to cultivated LSECs with an EC50 of approximately 22 µM. The cytochrome P450 (CYP)-dependency of senecionine bioactivation was confirmed in CYP reductase-deficient mice where no PA-induced hepatotoxicity was observed. Therefore, toxic metabolites of senecionine are generated by hepatic CYPs, and may be partially released from hepatocytes leading to destruction of LSECs in the pericentral region of the liver lobules. Analysis of hepatic bile salt transport by intravital two-photon imaging revealed a delayed uptake of a fluorescent bile salt analogue from the hepatic sinusoids into hepatocytes and delayed elimination. This was accompanied by transcriptional deregulation of hepatic bile salt transporters like Abcb11 or Abcc1. In conclusion, senecionine destroys LSECs although the toxic metabolite is formed in a CYP-dependent manner in the adjacent pericentral hepatocytes.

Original languageEnglish
Number of pages11
JournalArchives of Toxicology
Early online date12 Oct 2019
DOIs
Publication statusE-pub ahead of print - 12 Oct 2019

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Pyrrolizidine Alkaloids
Cholestasis
Endothelial cells
Liver
Cytochrome P-450 Enzyme System
Endothelial Cells
Poisons
Bile Acids and Salts
Hepatocytes
Cytotoxicity
Metabolites
Photons
NADPH-Ferrihemoprotein Reductase
Histology
Deregulation
senecionine
Platelets
Microscopic examination
Clinical Chemistry
Agglomeration

Keywords

  • Hepatotoxicity
  • Veno-occlusive disease
  • 2-Photon microscopy
  • Xenobiotic metabolism
  • Liver necrosis

Cite this

Hessel-Pras, Stefanie ; Braeuning, Albert ; Guenther, Georgia ; Adawy, Alshaimaa ; Enge, Anne-Margarethe ; Ebmeyer, Johanna ; Henderson, Colin J. ; Hengstler, Jan G. ; Lampen, Alfonso ; Reif, Raymond. / The pyrrolizidine alkaloid senecionine induces CYP-dependent destruction of sinusoidal endothelial cells and cholestasis in mice. In: Archives of Toxicology. 2019.
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abstract = "Pyrrolizidine alkaloids (PAs) are widely occurring phytotoxins which can induce severe liver damage in humans and other mammalian species by mechanisms that are not fully understood. Therefore, we investigated the development of PA hepatotoxicity in vivo, using an acutely toxic dose of the PA senecionine in mice, in combination with intravital two-photon microscopy, histology, clinical chemistry, and in vitro experiments with primary mouse hepatocytes and liver sinusoidal endothelial cells (LSECs). We observed pericentral LSEC necrosis together with elevated sinusoidal marker proteins in the serum of senecionine-treated mice and increased sinusoidal platelet aggregation in the damaged tissue regions. In vitro experiments showed no cytotoxicity to freshly isolated LSECs up to 500 µM senecionine. However, metabolic activation of senecionine by preincubation with primary mouse hepatocytes increased the cytotoxicity to cultivated LSECs with an EC50 of approximately 22 µM. The cytochrome P450 (CYP)-dependency of senecionine bioactivation was confirmed in CYP reductase-deficient mice where no PA-induced hepatotoxicity was observed. Therefore, toxic metabolites of senecionine are generated by hepatic CYPs, and may be partially released from hepatocytes leading to destruction of LSECs in the pericentral region of the liver lobules. Analysis of hepatic bile salt transport by intravital two-photon imaging revealed a delayed uptake of a fluorescent bile salt analogue from the hepatic sinusoids into hepatocytes and delayed elimination. This was accompanied by transcriptional deregulation of hepatic bile salt transporters like Abcb11 or Abcc1. In conclusion, senecionine destroys LSECs although the toxic metabolite is formed in a CYP-dependent manner in the adjacent pericentral hepatocytes.",
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author = "Stefanie Hessel-Pras and Albert Braeuning and Georgia Guenther and Alshaimaa Adawy and Anne-Margarethe Enge and Johanna Ebmeyer and Henderson, {Colin J.} and Hengstler, {Jan G.} and Alfonso Lampen and Raymond Reif",
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The pyrrolizidine alkaloid senecionine induces CYP-dependent destruction of sinusoidal endothelial cells and cholestasis in mice. / Hessel-Pras, Stefanie (Lead / Corresponding author); Braeuning, Albert; Guenther, Georgia; Adawy, Alshaimaa; Enge, Anne-Margarethe; Ebmeyer, Johanna; Henderson, Colin J.; Hengstler, Jan G.; Lampen, Alfonso; Reif, Raymond.

In: Archives of Toxicology, 12.10.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - The pyrrolizidine alkaloid senecionine induces CYP-dependent destruction of sinusoidal endothelial cells and cholestasis in mice

AU - Hessel-Pras, Stefanie

AU - Braeuning, Albert

AU - Guenther, Georgia

AU - Adawy, Alshaimaa

AU - Enge, Anne-Margarethe

AU - Ebmeyer, Johanna

AU - Henderson, Colin J.

AU - Hengstler, Jan G.

AU - Lampen, Alfonso

AU - Reif, Raymond

N1 - This work was supported by the German Research Foundation (Grant Number LA1177/12-1), by the German Federal Institute for Risk Assessment (Grant Numbers 1322-591 and 1322-624) and the BMBF Funded project LiSyM. ERL mice were generated under Cancer Research UK Programme Grant C4639/A10822 awarded to Professor C.R. Wolf.

PY - 2019/10/12

Y1 - 2019/10/12

N2 - Pyrrolizidine alkaloids (PAs) are widely occurring phytotoxins which can induce severe liver damage in humans and other mammalian species by mechanisms that are not fully understood. Therefore, we investigated the development of PA hepatotoxicity in vivo, using an acutely toxic dose of the PA senecionine in mice, in combination with intravital two-photon microscopy, histology, clinical chemistry, and in vitro experiments with primary mouse hepatocytes and liver sinusoidal endothelial cells (LSECs). We observed pericentral LSEC necrosis together with elevated sinusoidal marker proteins in the serum of senecionine-treated mice and increased sinusoidal platelet aggregation in the damaged tissue regions. In vitro experiments showed no cytotoxicity to freshly isolated LSECs up to 500 µM senecionine. However, metabolic activation of senecionine by preincubation with primary mouse hepatocytes increased the cytotoxicity to cultivated LSECs with an EC50 of approximately 22 µM. The cytochrome P450 (CYP)-dependency of senecionine bioactivation was confirmed in CYP reductase-deficient mice where no PA-induced hepatotoxicity was observed. Therefore, toxic metabolites of senecionine are generated by hepatic CYPs, and may be partially released from hepatocytes leading to destruction of LSECs in the pericentral region of the liver lobules. Analysis of hepatic bile salt transport by intravital two-photon imaging revealed a delayed uptake of a fluorescent bile salt analogue from the hepatic sinusoids into hepatocytes and delayed elimination. This was accompanied by transcriptional deregulation of hepatic bile salt transporters like Abcb11 or Abcc1. In conclusion, senecionine destroys LSECs although the toxic metabolite is formed in a CYP-dependent manner in the adjacent pericentral hepatocytes.

AB - Pyrrolizidine alkaloids (PAs) are widely occurring phytotoxins which can induce severe liver damage in humans and other mammalian species by mechanisms that are not fully understood. Therefore, we investigated the development of PA hepatotoxicity in vivo, using an acutely toxic dose of the PA senecionine in mice, in combination with intravital two-photon microscopy, histology, clinical chemistry, and in vitro experiments with primary mouse hepatocytes and liver sinusoidal endothelial cells (LSECs). We observed pericentral LSEC necrosis together with elevated sinusoidal marker proteins in the serum of senecionine-treated mice and increased sinusoidal platelet aggregation in the damaged tissue regions. In vitro experiments showed no cytotoxicity to freshly isolated LSECs up to 500 µM senecionine. However, metabolic activation of senecionine by preincubation with primary mouse hepatocytes increased the cytotoxicity to cultivated LSECs with an EC50 of approximately 22 µM. The cytochrome P450 (CYP)-dependency of senecionine bioactivation was confirmed in CYP reductase-deficient mice where no PA-induced hepatotoxicity was observed. Therefore, toxic metabolites of senecionine are generated by hepatic CYPs, and may be partially released from hepatocytes leading to destruction of LSECs in the pericentral region of the liver lobules. Analysis of hepatic bile salt transport by intravital two-photon imaging revealed a delayed uptake of a fluorescent bile salt analogue from the hepatic sinusoids into hepatocytes and delayed elimination. This was accompanied by transcriptional deregulation of hepatic bile salt transporters like Abcb11 or Abcc1. In conclusion, senecionine destroys LSECs although the toxic metabolite is formed in a CYP-dependent manner in the adjacent pericentral hepatocytes.

KW - Hepatotoxicity

KW - Veno-occlusive disease

KW - 2-Photon microscopy

KW - Xenobiotic metabolism

KW - Liver necrosis

U2 - 10.1007/s00204-019-02582-8

DO - 10.1007/s00204-019-02582-8

M3 - Article

C2 - 31606820

JO - Archives of Toxicology

JF - Archives of Toxicology

SN - 0340-5761

ER -