Does prostaglandin-E1 modulate D-galactosamine induced cell death in primary culture of human hepatocytes?
Background: Cell death pathway can occur under physiological or pathological conditions. In vitro and in vivo studies have shown that D-galactosamine (DGA) induces hepatocyte damage.
Objective: The present study aims to evaluate the protective effect of prostaglandin E1 (PGE1) on DGA-induced apoptosis, necrosis and oxidative stress in primary culture of human hepatocytes.
Methods: Normal human hepatocytes were obtained from the safety margin of liver specimens, removed during hepatectomy operation to liver cancer patients, and isolated using the classical collagenase perfusion method. After culture stabilization, PGE1 (1µM) was added 2 h before DGA (5 mM). Cultures were maintained for 24 h before the parameters for apoptosis, necrosis and oxidative stress were measured. Apoptosis was studied by DNA-fragmentation, neutral (nSMase) and acid (aSMase) sphingomyelinase and caspase-3 activity. Necrosis was investigated by lactate dehydrogenase (LDH) and transaminases (ALT & AST) enzymes. The oxidative stress was assessed by malondialdehyde (MDA), glutathione (GSH), oxidized glutathione (GSSH), glutathione S-transferase (GST), glutathione peroxidase (GSPx), catalase (CAT), superoxide dismutase (SOD) and nitric oxide (NO). Results: The hepatotoxin DGA induced apoptosis and enhanced all parameters related to necrosis and intracellular oxidative stress. On the other hand, PGE1 reduced the measured values for the parameters indicative for the DAG induced apoptosis, necrosis and oxidative stress. In addition, PGE1 proved also to prevent GSH depletion. The obtained results provided evidences for the biochemical hepatotoxic effects of DGA (5 mM) especially through the induction of apoptosis,necrosis and oxidative stress alterations in the cultured human hepatocytes.
Conclusion: PGE1 could be a useful protective treatment against DGA-induced hepatocyte cell death.
KEYWORDS Prostaglandin-E1; D-Galactosamine; Hepatocyte injury; Apoptosis; Necrosis; Oxidative stress