Cytochrome-P450 2B1 gene silencing attenuates puromycin aminonucleoside-induced cytotoxicity in glomerular epithelial cells.
Previously, we demonstrated that cytochrome P450 2B1 (CYP2B1) can generate reactive oxygen species in puromycin aminonucleoside (PAN)-induced nephrotic syndrome, an animal model of minimal-change disease in humans. In this study we found that overexpression of CYP2B1 in rat glomerular epithelial cells in vitro significantly increased PAN-induced reactive oxygen species generation, cytotoxicity, cell death, and collapse of the actin cytoskeleton. All of these pathological changes were markedly attenuated by siRNA-induced CYP2B1 silencing. The cellular CYP2B1 protein content was significantly decreased whereas its mRNA level was markedly increased, suggesting regulation by protein degradation rather than transcriptional inhibition in the PAN-treated glomerular epithelial cells. This degradation of CYP2B1 was accompanied by the induction of heme oxygenase-1, an important indicator of heme-induced oxidative stress. In PAN-treated CYP2B1-silenced glomerular epithelial cells the induction of heme oxygenase-1 and caspase-3 activity were significantly decreased. Further, cleavage of the stress-induced pro-apoptotic endoplasmic reticulum-specific pro-caspase-12 was prevented in the silenced cells. Our results support a pivotal role of CYP2B1 for reactive oxygen species production in the endoplasmic reticulum in PAN-induced cytotoxicity.
Tian
Niu
N
Department of Pediatrics, University of Mississippi Medical Center, Jackson, Mississippi 39216, USA.
Arany
Istvan
I
Waxman
David J
DJ
Baliga
Radhakrishna
R
D000818:Animals; D002478:Cells, Cultured; D019362:Cytochrome P-450 CYP2B1 / Q000096:biosynthesis / Q000235:genetics*; D004847:Epithelial Cells / Q000187:drug effects*; D015870:Gene Expression; D020868:Gene Silencing; D007678:Kidney Glomerulus / Q000166:cytology* / Q000187:drug effects*; D011692:Puromycin Aminonucleoside / Q000633:toxicity*; D051381:Rats
D011692:Puromycin Aminonucleoside; D019362:Cytochrome P-450 CYP2B1
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