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Establishment of a novel CYP3A4-transduced human hepatic sinusoidal endothelial cell model and its application in screening hepatotoxicity of pyrrolizidine alkaloids
- Source :
- Journal of environmental science and health. Part C, Toxicology and carcinogenesis. 38(2)
- Publication Year :
- 2020
-
Abstract
- Pyrrolizidine alkaloids (PAs) are extensively distributed in plants and are known to damage hepatic sinusoidal endothelial cells (HSECs) via metabolic activation mediated by hepatic cytochrome P450 enzymes (CYPs), particularly the CYP3A4 isozyme. Different PAs have distinct toxic potencies and their toxic effects on HSECs are difficult to be determined in cultured cells, because HSECs lack the key CYP3A4 isozyme for metabolic activation. This study aims to establish a novel, convenient and reliable CYP3A4-expressing HSEC model using human HSECs transduced with lentivirus carrying CYP3A4-ires-eGFP, for evaluating the hepatotoxicity of different PAs on their target HSECs. The developed CYP3A4-expressing HSEC (HSEC-CYP3A4) model was verified by the expression of GFP and CYP3A4 and by the ability to metabolize nifedipine, a classic CYP3A4 substrate. Treated with retrorsine, a representative toxic PA, HSEC-CYP3A4 cells showed significantly reduced cell viability, depletion of GSH, and increased formation of pyrrole-protein adducts. Furthermore, this newly developed cell model successfully discriminated the cytotoxic potency of different PAs evidenced by their IC40 values. In conclusion, the established HSEC-CYP3A4 cell model can be used as a rapid screening platform for assessing the relative potencies of individual PAs on their target HSECs and for investigating the mechanisms underlying PA-induced hepatic sinusoidal damage.
- Subjects :
- Cancer Research
Cell Survival
Health, Toxicology and Mutagenesis
Acinar Cells
Pharmacology
Hepatic Veins
Isozyme
Green fluorescent protein
Cell Line
Activation, Metabolic
chemistry.chemical_compound
Cytochrome P-450 Enzyme System
Cytochrome P-450 CYP3A
Humans
Viability assay
Mononuclear Phagocyte System
Cells, Cultured
Pyrrolizidine Alkaloids
CYP3A4
biology
Endothelial Cells
Glutathione
biology.organism_classification
Endothelial stem cell
chemistry
Liver
Pyrrolizidine
Lentivirus
Chemical and Drug Induced Liver Injury
Subjects
Details
- ISSN :
- 26896591
- Volume :
- 38
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Journal of environmental science and health. Part C, Toxicology and carcinogenesis
- Accession number :
- edsair.doi.dedup.....0d7ccb84c0488e40149e2657046de60a