Mechanisms of tolvaptan-induced toxicity in HepG2 cells
Open accessAbstract
Tolvaptan, a vasopressin receptor 2 antagonist used to treat hyponatremia, has recently been reported to be associated with an increased risk of liver injury. In this study, we explored the underlying mechanisms of hepatotoxicity of tolvaptan using human HepG2 cells. Tolvaptan inhibited cell growth and caused cell death in a concentration- and time-dependent manner. Tolvaptan treatment led to delayed cell cycle progression, accompanied by decreased levels of several cyclins and cyclin-dependent kinases. Tolvaptan was found to cause DNA damage, as assessed by alkaline comet assays; this was confirmed by increased levels of 8-oxoguanine and phosphorylation of histone H2AX. Exposure of HepG2 cells to tolvaptan enhanced cytochrome C release and triggered apoptosis by modulating Bcl-2 family members. The activation of p38 contributed to tolvaptan-mediated apoptosis via down-regulation of Bcl-2. Proteasome inhibition altered tolvaptan-induced cell cycle deregulation and enhanced tolvaptan-induced apoptosis and cytotoxicity. Moreover, tolvaptan treatment induced autophagy. Inhibition of autophagy by knocking-down an autophagy-related gene increased tolvaptan-induced apoptosis and cytotoxicity. Taken together, our findings suggest that the cytotoxicity of tolvaptan results from delayed cell cycle progression, the induction of DNA damage, and the execution of apoptosis. In addition, a number of signaling pathways were perturbed by tolvaptan and played an important role in its cytotoxicity.
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HepG2 has sharply reduced expression of Phase I CYP450 enzymes compared to primary human hepatocytes, with mRNA levels of key isoforms hundreds of times lower.
“HepG2 is characterized by a sharply reduced expression of Phase I biotransformation enzymes— CYP450 —compared to primary human hepatocytes [102,103,104,105].”
From The HepG2 Cell Line as a Model for Studying Metabolic Dysfunction-Associated Steatotic Liver Disease (2026) · cites this paper as [104]
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- Authors
- Wu, Y., Beland, F. A., Chen, S., Liu, F., Guo, L., & Fang, J.-L.
- Affiliations
- National Center for Toxicological Research (US)
- Corresponding author
- Fang, Jia-Long
- Venue
- Biochemical Pharmacology
- Publisher
- Elsevier BV
- Published
- 2015-06
- Volume
- 95
- Issue
- 4
- Pages
- 324-336
- ISSN
- 0006-2952 (ISSN-L)
- PMC
- PMC5894093
- OpenAlex
- W2024407138
- Cited by
- 41
- References
- 72
- Citations by year
- 2026: 3 · 2025: 3 · 2024: 4 · 2023: 2 · 2022: 4 · 2021: 5 · 2020: 2 · 2019: 3 · 2017: 6 · 2016: 5 · 2015: 3 · 2014: 1
- FWCI
- 5.44
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Biochemical Pharmacology (acceptedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
- Funders
- U.S. Department of Energy
- FDA
- Topics
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Toxicology › Bioactive Compounds and Antitumor Agents
- Keywords
- tolvaptan, hepatotoxicity, drug-induced liver injury, HepG2 cells, apoptosis, cell cycle arrest, DNA damage, 8-oxoguanine, H2AX phosphorylation, cytochrome c release, Bcl-2 family proteins, p38 MAPK, proteasome inhibition, autophagy, cytotoxicity
- MeSH terms
- Tolvaptan, Autophagy, Benzazepines, Cell Cycle, DNA Damage, Enzyme Activation, Humans, Cell Death, Apoptosis, Mitogen-Activated Protein Kinases, Proteasome Endopeptidase Complex, Cell Proliferation, Cyclin-Dependent Kinase 4, Cytochrome P-450 CYP3A, Cyclin D3, Hep G2 Cells, Proteasome Inhibitors, Antidiuretic Hormone Receptor Antagonists
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