| Inhibitors directed towards caspase-1 and -3 are less effective than pan caspase inhibition in preventing renal proximal tubular cell apoptosis. | |
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MedLine Citation:
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PMID: 14988591 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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BACKGROUND: Uncontrolled apoptosis contributes to tubular cell deletion in renal scarring. Caspase-3 has a central role in the execution of apoptosis and may provide a target for regulating cell death. Here we evaluate three caspase inhibitors: B-D-FMK (pan caspase inhibitor), Z-DEVDFMK (predominantly Caspase-3 inhibitor) and Z-VAD-FMK (predominantly Caspase-1 and -3 inhibitor) to ameliorate apoptosis induced by cisplatin in rat proximal tubular (RPT) cells. METHODS: Caspase-3 activity (substrate cleavage assay) and protein (immunocytochemistry and Western blotting), apoptosis (Annexin V flow cytometry, in situend labelling of fragmented DNA, light/electron microscopy and DNA laddering) and cell survival (trypan blue exclusion and propidium iodide flow cytometry) were determined in RPT cells exposed to cisplatin with and without caspase inhibitors. RESULTS: Cisplatin induced a dose-dependent increase in Caspase-3 activity and 8-fold of increase in apoptosis (p < 0.01) when applied for 24 h at 100 microM. B-D-FMK (40 microM), Z-DEVD-FMK (15 microM) and Z-VAD-FMK (22 microM) almost completely inhibited the 25-fold increase in Caspase-3 activity and decreased apoptosis from 15.9 +/- 4.4 to 2.0 +/- 0.6% (p < 0.01), 15.0 +/- 2.2 and 15.0 +/- 2.2% respectively. DNA ladders were visible in cisplatin-treated cells, which disappeared following addition of B-D-FMK and decreased with Z-VAD-FMK and Z-DEVD-FMK. Cisplatin reduced cell survival to 61% by trypan blue exclusion. B-D-FMK and Z-VAD-FMK increased this to 87 and 75%, but Z-DEVD-FMK had no significant effect. CONCLUSIONS: Cisplatin causes an increase in RPT apoptosis that is associated with increased Caspase-3 activity. All caspase inhibitors were equally effective at reducing Caspase-3 activity, however the pan caspase inhibitor B-D-FMK was more effective at preventing apoptosis and increasing cell survival. Therefore, pan caspase inhibition offers the greatest potential for the prevention of renal tubular cell deletion by uncontrolled apoptosis. |
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Authors:
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Bin Yang; A Meguid El Nahas; Marie Fisher; Bart Wagner; Linhong Huang; Ian Storie; David Barnett; Jonathan Barratt; Alice C Smith; Timothy S Johnson |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Nephron. Experimental nephrology Volume: 96 ISSN: 1660-2129 ISO Abbreviation: Nephron Exp. Nephrol. Publication Date: 2004 |
Date Detail:
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Created Date: 2004-02-27 Completed Date: 2004-05-10 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 101159770 Medline TA: Nephron Exp Nephrol Country: Switzerland |
Other Details:
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Languages: eng Pagination: e39-51 Citation Subset: IM |
Copyright Information:
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Copyright 2004 S. Karger AG, Basel |
Affiliation:
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Sheffield Kidney Institute, Sheffield University Division of Clinical Sciences, Sheffield, UK. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Chloromethyl Ketones
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pharmacology Animals Apoptosis / drug effects* Caspase 1 / antagonists & inhibitors Caspase 3 Caspases / analysis, antagonists & inhibitors*, metabolism Cell Line Cell Survival / drug effects Cisplatin / antagonists & inhibitors Cysteine Proteinase Inhibitors / pharmacology* DNA Fragmentation / drug effects Flow Cytometry Kidney Tubules, Proximal / cytology*, drug effects, enzymology Necrosis Oligopeptides / pharmacology Rats |
| Chemical | |
Reg. No./Substance:
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0/Amino Acid Chloromethyl Ketones; 0/Cysteine Proteinase Inhibitors; 0/Oligopeptides; 0/benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone; 0/benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; 0/butyloxycarbonyl-O-methyl-aspartyl-fluoromethyl ketone; 15663-27-1/Cisplatin; EC 3.4.22.-/Casp3 protein, rat; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspases; EC 3.4.22.36/Caspase 1 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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