PMID- 8529915
OWN - NLM
STAT- MEDLINE
DCOM- 19960201
LR  - 20190825
IS  - 0891-5849 (Print)
IS  - 0891-5849 (Linking)
VI  - 19
IP  - 5
DP  - 1995 Nov
TI  - Cellular injury induced by oxidative stress is mediated through lysosomal damage.
PG  - 565-74
AB  - Cultured primary hepatocytes pretreated (protected) with the iron chelator 
      deferoxamine or the antioxidant N,N'-diphenyl-p-phenylenediamine (DPPD) were 
      resistant to the toxicity of 5 microM naphthazarin 
      (5,8-dihydroxy-1,4-naphthoquinone) during a 180-min exposure. Hepatocytes exposed 
      to naphthazarin without any protection were abruptly depleted of intracellular 
      reduced glutathione, and the level of cytosolic Ca2+ was rapidly increased. This 
      was followed by lipid peroxidation, measured as accumulation of malondialdehyde 
      (MDA) and 4-hydroxyalkenals (4-HNA) intra- and extracellularly; decrease in ATP 
      levels; destabilization of lysosomes; and finally cell death. The stability of 
      the lysosomal membranes was evaluated by determining retention of the 
      lysosomotropic weak base acridine orange (AO). Naphthazarin exposure caused 
      leakage of protons from the acidic compartment, as indicated by relocalization of 
      AO to the cytosol. Protection of the cell cultures with deferoxamine or DPPD 
      prevented destabilization of lysosomes and cell killing. It also reduced the loss 
      of ATP but did not prevent the depletion of glutathione or the increase in Ca2+. 
      In cells subjected to naphthazarin exposure, DPPD protection also completely 
      inhibited lipid peroxidation, whereas deferoxamine pretreatment only slightly 
      reduced the intracellular accumulation of MDA and 4-HNA but completely prevented 
      cell rupture and the leakage of these lipid peroxidation products to the medium 
      that took place in large amounts from unprotected cells exposed to naphthazarin. 
      Deferoxamine is taken up by endocytosis and is thus transported to the acidic 
      vacuolar apparatus, whereas the lipophilic DPPD is rapidly distributed throughout 
      the cells. Inhibiting endocytosis during deferoxamine pretreatment, by incubating 
      at +4 degrees C or by preexposure to a mixture of the endocytosis-inhibitors 
      cytochalasin B and monensin, abolished the protective effect of deferoxamine. The 
      findings suggest that naphthazarin-induced cell killing is not caused directly by 
      either thiol oxidation or an increase in cytosolic free Ca2+, but rather is 
      preceded by lysosomal destabilization, which may be prevented either by 
      inhibition of cellular peroxidation in general or by prevention of iron-catalyzed 
      oxidative reactions, and involves peroxidation of cellular membranes, energy 
      depletion, and leakage of lysosomal content. DPPD would protect against cell 
      killing by preventing lipid peroxidation of cellular membranes in general, 
      whereas deferoxamine seems to allow a limited general cellular peroxidation but 
      specifically prevents peroxidation and fragmentation of lysosomal membranes by 
      chelating intralysosomal iron and, consequently, leakage of destructive lysosomal 
      contents with ensuing cell rupture and death. Thus, a certain degree of cellular 
      peroxidation does not appear to be lethal as long as lysosomal membranes are 
      protected, placing lysosomes into a category of cellular loci minora resistentia.
FAU - Ollinger, K
AU  - Ollinger K
AD  - Department of Pathology II, Faculty of Health Sciences, University of Linkoping, 
      Sweden.
FAU - Brunk, U T
AU  - Brunk UT
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Free Radic Biol Med
JT  - Free radical biology & medicine
JID - 8709159
RN  - 0 (Antineoplastic Agents)
RN  - 0 (Antioxidants)
RN  - 0 (Iron Chelating Agents)
RN  - 0 (Naphthoquinones)
RN  - 0 (Phenylenediamines)
RN  - 3CHI920QS7 (Cytochalasin B)
RN  - 475-38-7 (naphthazarin)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - 906O0YJ6ZP (Monensin)
RN  - DD517SCM92 (N,N'-diphenyl-4-phenylenediamine)
RN  - GAN16C9B8O (Glutathione)
RN  - J06Y7MXW4D (Deferoxamine)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Adenosine Triphosphate/metabolism
MH  - Animals
MH  - Antineoplastic Agents/antagonists & inhibitors/*toxicity
MH  - Antioxidants/pharmacology
MH  - Calcium/metabolism
MH  - Cell Survival/drug effects
MH  - Cells, Cultured
MH  - Cytochalasin B/pharmacology
MH  - Cytosol/metabolism
MH  - Deferoxamine/pharmacology
MH  - Endocytosis/drug effects
MH  - Glutathione/metabolism
MH  - Iron Chelating Agents/pharmacology
MH  - Kinetics
MH  - Liver/drug effects/*pathology/physiology
MH  - Lysosomes/drug effects/*pathology/physiology
MH  - Male
MH  - Monensin/pharmacology
MH  - Naphthoquinones/antagonists & inhibitors/*toxicity
MH  - *Oxidative Stress
MH  - Phenylenediamines/pharmacology
MH  - Rats
MH  - Rats, Wistar
MH  - Time Factors
EDAT- 1995/11/01 00:00
MHDA- 1995/11/01 00:01
CRDT- 1995/11/01 00:00
PHST- 1995/11/01 00:00 [pubmed]
PHST- 1995/11/01 00:01 [medline]
PHST- 1995/11/01 00:00 [entrez]
AID - 0891584995000623 [pii]
AID - 10.1016/0891-5849(95)00062-3 [doi]
PST - ppublish
SO  - Free Radic Biol Med. 1995 Nov;19(5):565-74. doi: 10.1016/0891-5849(95)00062-3.