In vivo inhibition by antioxidants of adriamycin-induced apoptosis in murine peritoneal macrophages.
Adriamycin (ADM) is an oncostatic of the anthracycline family with confirmed experimental and clinical efficiency. This antitumoral drug has been reported to stimulate macrophage activity and is able to induce apoptosis (AP) in some tumour cells. The objective of the present work was to investigate if in vivo administration of ADM to mice induces AP in their peritoneal macrophages (PM). AP was expressed by the apoptotic index (AI) of peritoneal macrophages observed under fluorescence microscope after ethidium bromide and acridine orange staining and confirmed by detection of the ladder pattern on DNA electrophoresis, indicates DNA fragmentation in 80-120 bp characteristic of apoptotic state. 24 hours after i.p. ADM administration, AP was observed in PM. The effect was best visible after the injection of 5 mg/kg ADM. (Al: 76.3+/-8.9 vs untreated control group AI: 2.8+/-1.1). In the ADM treated group a DNA ladder electrophoretic pattern was observed while DNA from normal PM was genomic. Since ADM toxicity has been attributed to reactive oxygen species generation, we investigated its possible participation in AP induction by pretreating mice with antioxidants: (+)-alpha-tocopherol acid succinate (30 IU/mouse per os) for 3 days before ADM administration with E. coli lipopolysacharide (0.15 microg/mouse i.p.) 24 hours before ADM administration or with superoxide dismutase (10,000 IU/mouse i.p.) 1 hour before ADM administration. AI was significantly decreased, with values close to those of the untreated control group (AI: 15+/-5.7, 9.6+/-8.0 and 32.9+/-6.9, respectively). Antioxidants given before ADM treatment significantly increased the live cell index (p < or = 0.001) in PM the groups while inactivated antioxidants no longer protect PM against the ADM AP induction. DNA analysis confirmed the effect: in the untreated control and in the antioxidant protected groups DNA was genomic while in either ADM or inactivated-antioxidants + ADM treated groups, DNA presented the ladder pattern. AP can thus be induced in PM by ADM and inhibited by antioxidants. These observations may have clinical applications.
Dominguez-Rodriguez
J R
JR
Centro de Investigación Biomédica de Occidente, IMSS, C. U. C. S. Universidad de Guadalajara, Mexico.
Gomez-Contreras
P C
PC
Hernandez-Flores
G
G
Lerma-Diaz
J M
JM
Carranco
A
A
Cervantes-Munguia
R
R
Orbach-Arbouys
S
S
Bravo-Cuella
A
A
D000165:Acridine Orange / Q000494:pharmacology; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis; D002470:Cell Survival; D053938:DNA Fragmentation; D004317:Doxorubicin / Q000494:pharmacology*; D004996:Ethidium / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D017737:Macrophages, Peritoneal / Q000473:pathology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D017382:Reactive Oxygen Species; D013482:Superoxide Dismutase / Q000494:pharmacology; D024505:Tocopherols; D014810:Vitamin E / Q000031:analogs & derivatives* / Q000494:pharmacology
D000970:Antineoplastic Agents; D000975:Antioxidants; D005456:Fluorescent Dyes; D008070:Lipopolysaccharides; D017382:Reactive Oxygen Species; D014810:Vitamin E; D004317:Doxorubicin; D013482:Superoxide Dismutase; D004996:Ethidium; D000165:Acridine Orange; D024505:Tocopherols