In Vitro Induction of Apoptosis in Rat Hepatocytes by Cyclosporine A

摘要: In rat hepatocytes and isolated liver mitochondrial fractions, CsA is often used as a specific inhibitor of Ca2+ release blocker membrane potential permeability transition (MPT), which are all processes involved in the inhibition apoptosis. However, neither nor induction apoptosis by has yet been described hepatocyte primary culture during incubation for 4 20 hours. It was purpose present study to examine means morphological biochemical criteria effects on apoptosis, characterize underlying mechanisms. Rat were cultured or hours with at concentrations 0, 10, 25 50 ?M. Chromatin condensation fragmentation, DNA fragmentation (TUNEL), phosphatidylserine distribution (Annexin V), caspase-1, -3 -6 activity, (Rhodamine 123), cytochrome c into cytosol investigated. Four after treatment, chromatin number TUNEL- Annexin V-positive cells increased dose dependentlywithout any observable enzyme leakage, indicated integrity outer cell membrane. After parameters further accompanied activity cysteine protease, caspase-3 (CPP 32), slightly caspase-6 (Mch 2), but not caspase-1 (ICE). The inhibitor, Ac-DEVD-CHO, inhibited activation attenuated CsA-induced LDH leakage. Ac-VEIDCHO, only marginally Decreased went parallel ultrastructural changes might be regarded early events trigger cascade. Transmission electron microscopy (TEM) confirmed an increase necrotic hours, compared controls.