Puromycin Aminonucleoside Induces Glomerular Epithelial Cell Apoptosis

• Published in: Experimental and molecular pathology 2001-02, Vol.70 (1), p.54-64
• Main Authors: Sanwal, Vibha, Pandya, Manish, Bhaskaran, Madhu, Franki, Nicholas, Reddy, Krishna, Ding, Guohua, Kapasi, Aditi, Valderrama, Elsa, Singhal, Pravin C.
• Format: Article
• Language: English
• Subjects: actinomycin D; Animals; Apoptosis - drug effects; Biological and medical sciences; Catalase - pharmacology; Cells, Cultured; Dactinomycin - pharmacology; Disease Models, Animal; DNA - drug effects; DNA Fragmentation - drug effects; Dose-Response Relationship, Drug; Electrophoresis, Agar Gel; Epithelial Cells - drug effects; Epithelial Cells - pathology; free radical scavengers; Free Radical Scavengers - pharmacology; Glomerulonephritis; glomerulosclerosis; Humans; In Situ Nick-End Labeling; Kidney Glomerulus - drug effects; Kidney Glomerulus - pathology; Medical sciences; mesangium; necrosis; Nephrology. Urinary tract diseases; Nephropathies. Renovascular diseases. Renal failure; Puromycin Aminonucleoside - toxicity; Rats; Rats, Sprague-Dawley; Superoxide Dismutase - pharmacology; Thiourea - analogs & derivatives; Thiourea - pharmacology
• ISSN: 0014-4800; 1096-0945
• DOI: 10.1006/exmp.2000.2345

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 μg/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 μg/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.