Biochemical characterisation of an in vitro model of hepatocellular apoptotic cell death.

摘要: This study was set up to critically evaluate a commonly-used in vitro model of hepatocellular apoptotic cell death, which freshly isolated hepatocytes, cultured monolayer configuration, are exposed combination Fas ligand and cycloheximide for six hours. A well-acknowledged death markers addressed: a) morphology studied by light microscopy; b) necrotic populations were quantified situ staining with Annexin-V, Hoechst 33342 propidium iodide (PI); c) activities monitored probing caspase 3-like activity measuring the extracellular leakage lactate dehydrogenase (LDH), respectively; d) expression apoptosis regulators investigated immunoblotting. The initiation evidenced activation 8 9, increased Annexin-V reactivity. Progression through process confirmed 3 Bid, enhanced Bax, occurrence nuclear fragmentation. Late transition appearance demonstrated an number PI-positive cells augmented release LDH. Thus, allows entire course Fas-mediated is not possible vivo. experimental system can serve broad range pharmaco-toxicological purposes, thereby directly assisting reduction animal experimentation.