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Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205
The goals of this study were to determine 1) the expression of calpain isoforms in rabbit renal proximal tubules (RPT); 2) calpain autolysis and translocation, and calpastatin levels during RPT injury; and 3) the effect of a calpain inhibitor (PD-150606) on calpain levels, mitochondrial function, and ion transport during RPT injury. RT-PCR, immunoblot analysis, and FITC-casein zymography demonstrated the presence of only µ- and m-calpains in rabbit RPT. The mitochondrial inhibitor antimycin A decreased RPT µ- and m-calpain and calpastatin levels in conjunction with cell death and increased plasma membrane permeability. No increases in either µ- or m-calpain were observed in the membrane nor were increases observed in autolytic forms of either µ- or m-calpain in antimycin A-exposed RPT. PD-150606 blocked antimycin A-induced cell death, preserved calpain levels in antimycin A-exposed RPT, and promoted the recovery of mitochondrial function and active Na+ transport in RPT after hypoxia and reoxygenation. The present study suggests that calpains mediate RPT injury without undergoing autolysis or translocation, and ultimately they leak from cells subsequent to RPT injury/death. Furthermore, PD-150606 allows functional recovery after injury.
rabbit; mitochondrial inhibitor; hypoxia/reoxygenation; mitochondrial function; active sodium transport; calpastatin
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