Stefano Fiorucci1, Elisabetta Antonelli1, Eleonora Distrutti1, Andrea Mencarelli1, Silvana Farneti1, Piero Del Soldato2, Antonio Morelli1
1Dipartimento di Medicina Clinica e Sperimentale, Clinica di Gastroenterologia ed Epatologia, Università degli Studi di Perugia, Italy.
2Nicox, SA, Sophia Antipolis, France
Tóm tắt
NCX‐1000, (3α, 5β, 7β)‐3,7‐dihydroxycholan‐24oic acid[2‐methoxy‐4‐[3‐[4‐(nitroxy)butoxy]‐3‐oxo‐1‐propenyl]phenyl ester, is a nitric oxide (NO)‐derivative of ursodeoxyxholic acid (UDCA) that selectively release NO in the liver.
Here, we demonstrated that administering mice with 40 μmol kg−1 NCX‐1000, but not UDCA, improves liver histopathology and reduces mortality caused by 330 μmol kg−1 APAP from 60 to 25% (P<0.01). Administration of NCX‐1000, in a therapeutic manner, that is, 2 h after acetaminophen (APAP) intoxication reduced mortality, improved liver histopathology and prevented liver IFN‐γ, TNF‐α, Fas/Fas ligand and inducible nitric oxide synthase (iNOS) mRNA accumulation caused by APAP.
In vitro exposure of primary cultures of mouse hepatocytes to APAP, 6.6 mM, resulted in apoptosis followed by necrosis. Loss of cell viability correlates with early mitochondrial membrane potential (Δψm) hyperpolarization followed by depolarization and cytochrome c translocation from mitochondria to cytosol. APAP‐induced apoptosis associated with procaspase‐3 and ‐9 cleavage, appearance of truncated Bid and activation of poly(ADP‐ribose) polymerase (PARP).
Treating primary culture of hepatocytes with 5 μM cyclosporine and 10 μM trifluoperazine for eight resulted in significant reduction of apoptosis induced by APAP suggesting that loss of Δψm was mechanistically involved in apoptosis induced by APAP in vitro.
NCX‐1000, but not UDCA, concentration‐dependently (ED50=16 μM) protected against Δψm depolarization and reduced transition from apoptosis to necrosis caused by 6.6 mM APAP.
Treating primary cultures of hepatocytes with the NO‐donor DETA‐NO, 100 μM, reduced apoptosis induced by APAP and prevented caspase activation.
In conclusion, NCX‐1000 is effective in protecting against APAP‐induced hepatotoxicity when administered in a therapeutic manner. This protection may involve the inhibition of apoptosis and the maintenance of mitochondrial integrity.
British Journal of Pharmacology (2004) 143, 33–42. doi:10.1038/sj.bjp.0705780
