Phosphate is not an absolute requirement for the inhibitory effects ofTI Phosphate is not an absolute requirement for the inhibitory effects of cyclosporin A or cyclophilin D deletion on mitochondrial permeability transition

摘要

CypD (cyclophilin D) has been established as a critical regulator of the MPT (mitochondrial permeability transition) pore, and pharmacological or genetic inhibition of CypD attenuates MPT in numerous systems. However, it has recently been suggested that the inhibitory effects of CypD inhibition only manifest when P-i (inorganic phosphate) is present, and that inhibition is lost when P-i is replaced by As-i (inorganic arsenate) or V-i (inorganic vanadate). To test this, liver mitochondria were isolated from wild-type and CypD-deficient (Ppif(-/-)) mice and then incubated in buffer containing P-i, As-i or V-i. MPT was induced under both energized and de-energized conditions by the addition of Ca2+, and the resultant mitochondrial swelling was measured spectrophotometrically. For pharmacological inhibition of CypD, wild-type mitochondria were pre-incubated with CsA (cyclosporin A) before the addition of Ca2+ In energized and de-energized mitochondria, Ca2+ induced MPT regardless of the anion present, although the magnitude differed between P-i, As-i and V-i. However, in all cases, pre-treatment with CsA significantly inhibited MPT. Moreover, these effects were independent of mouse strain, organ type and rodent species. Similarly, attenuation of Ca2+-induced MPT in the Ppif(-/-) mitochondria was still observed irrespective of whether P-i, As-i or V-i was present. We conclude that the pharmacological and genetic inhibition of CypD is still able to attenuate MPT even in the absence of P-i.