Puromycin aminonucleoside induces glomerular epithelial cell apoptosis.
Glomerular epithelial cell (GEC) injury has been considered to play an important role in puromycin aminonucleoside (PAN)-induced nephrosis. We studied the effect of PAN on rat as well as human GEC apoptosis. Morphogic evaluation of GEC apoptosis and necrosis was carried out by staining with H-33342 and propidium iodide. GEC apoptosis was further confirmed by DNA fragmentation assay (by both agarose gel electrophoresis and end-labeling). To determine the dose- and time-response effect of PAN, GECs were treated with variable concentrations of PAN (10 to 500 microg/ml) for variable time periods (6 to 48 h). To determine the role of gene synthesis, we studied the effect of actinomycin D (a transcriptional inhibitor) on PAN-induced GEC apoptosis. To determine the role of free radicals, we evaluated the effect of superoxide dismutase (SOD), dimethylthiourea (DMTU), and catalase on PAN-induced GEC apoptosis. PAN induced GEC apoptosis in a dose- and time-dependent manner. PAN at a high concentration (PAN, 100 microg/ml) also induced a moderate degree of GEC necrosis. In DNA fragmentation assays PAN-treated GECs showed the classic ladder pattern. PAN-induced GEC apoptosis was partly attenuated with free radical scavengers, such as SOD, DMTU, and catalase. In addition, actinomycin D attenuated PAN-induced GEC apoptosis. PAN induces GEC apoptosis, which may be mediated through the generation of reactive oxygen species.
Sanwal
V
V
Molecular Biology and Experimental Pathology Section, Long Island Jewish Medical Center and North Shore University Hospital, New York, New York 11040, USA.
Pandya
M
M
Bhaskaran
M
M
Franki
N
N
Reddy
K
K
Ding
G
G
Kapasi
A
A
Valderrama
E
E
Singhal
P C
PC
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002374:Catalase / Q000494:pharmacology; D002478:Cells, Cultured; D004247:DNA / Q000187:drug effects; D053938:DNA Fragmentation / Q000187:drug effects; D003609:Dactinomycin / Q000494:pharmacology; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D004847:Epithelial Cells / Q000187:drug effects / Q000473:pathology; D016166:Free Radical Scavengers / Q000494:pharmacology; D006801:Humans; D020287:In Situ Nick-End Labeling; D007678:Kidney Glomerulus / Q000187:drug effects* / Q000473:pathology; D011692:Puromycin Aminonucleoside / Q000633:toxicity*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013482:Superoxide Dismutase / Q000494:pharmacology; D013890:Thiourea / Q000031:analogs & derivatives* / Q000494:pharmacology
D016166:Free Radical Scavengers; D003609:Dactinomycin; D011692:Puromycin Aminonucleoside; C038983:1,3-dimethylthiourea; D004247:DNA; D002374:Catalase; D013482:Superoxide Dismutase; D013890:Thiourea