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Diclofenac induces apoptosis in hepatocytes
Maria-José Gómez-Lechón1, Xavier Ponsoda, Enrique O'Connor
1Centro de Investigación, Hospital La Fe, Avda de Campanar 21, E-46009, Valencia, Spain. gomez_mjo@gva.es
Summary
Diclofenac induces apoptosis in rat hepatocytes by activating caspase 3 and opening the mitochondrial permeability transition (MPT) pore, driven by oxidative stress. This effect may involve CYP-mediated metabolism.
Area of Science:
- Hepatology
- Toxicology
- Cell Biology
Background:
- Diclofenac, a widely used NSAID, is known to cause hepatotoxicity.
- Understanding the mechanisms of diclofenac-induced liver injury is crucial for patient safety.
Purpose of the Study:
- To investigate the early apoptotic events triggered by diclofenac in rat hepatocytes.
- To elucidate the role of mitochondrial permeability transition (MPT) and oxidative stress in diclofenac-induced apoptosis.
Main Methods:
- Incubation of rat hepatocytes with varying concentrations of diclofenac.
- Flow cytometry to assess apoptotic nuclei and DNA content.
- Measurement of Caspase 3 activation and lactate dehydrogenase (LDH) leakage.
- Assessment of MPT using inhibitors (Cyclosporine A, decylubiquinone) and antioxidants.
Main Results:
- Diclofenac induced apoptosis at sub-cytotoxic concentrations (450 microM) without causing necrosis.
- A time- and dose-dependent increase in apoptotic nuclei and Caspase 3 activation (3-5 fold) was observed.
- Diclofenac induced MPT pore opening, which was inhibited by Cyclosporine A and decylubiquinone.
- Antioxidants prevented Caspase 3 activation, indicating oxidative stress involvement in MPT induction.
- Differential effects in hepatocytes versus hepatoma cells suggest a role for CYP-mediated metabolism.
Conclusions:
- Diclofenac triggers apoptosis in hepatocytes via MPT pore opening and oxidative stress.
- Caspase 3 activation is a key event in diclofenac-induced hepatocyte apoptosis.
- CYP-mediated metabolism might contribute to diclofenac's apoptotic effects.