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Puromycin aminonucleoside induces glomerular epithelial cell apoptosis
V Sanwal1, M Pandya, M Bhaskaran
1Molecular Biology and Experimental Pathology Section, Long Island Jewish Medical Center and North Shore University Hospital, New York, New York 11040, USA.
Experimental and Molecular Pathology
|February 15, 2001
Summary
Puromycin aminonucleoside (PAN) induces glomerular epithelial cell (GEC) apoptosis in a dose- and time-dependent manner, potentially through reactive oxygen species. Free radical scavengers and actinomycin D attenuated this PAN-induced GEC apoptosis.
Area of Science:
- Nephrology
- Cell Biology
- Toxicology
Background:
- Glomerular epithelial cell (GEC) injury is implicated in puromycin aminonucleoside (PAN)-induced nephrosis.
- Understanding the mechanisms of GEC injury is crucial for developing therapeutic strategies.
Purpose of the Study:
- To investigate the effect of PAN on GEC apoptosis in both rat and human cells.
- To elucidate the role of gene synthesis and free radicals in PAN-induced GEC apoptosis.
Main Methods:
- Morphological evaluation of GEC apoptosis and necrosis using H-33342 and propidium iodide staining.
- DNA fragmentation assays (agarose gel electrophoresis, end-labeling) to confirm apoptosis.
- Treatment with varying PAN concentrations and durations, actinomycin D, and free radical scavengers (SOD, DMTU, catalase).
Main Results:
- PAN induced GEC apoptosis in a dose- and time-dependent manner.
- High concentrations of PAN also caused moderate GEC necrosis.
- DNA fragmentation assays showed a classic ladder pattern in PAN-treated GECs.
- Free radical scavengers and actinomycin D partially attenuated PAN-induced GEC apoptosis.
Conclusions:
- PAN induces GEC apoptosis, suggesting a significant role in PAN-induced nephrosis.
- The mechanism of PAN-induced GEC apoptosis may involve the generation of reactive oxygen species.
- Inhibition of gene synthesis also plays a role in mitigating PAN-induced GEC apoptosis.