Toninello Antonio, Salvi Mauro, Schweizer Matthias, Richter Christoph
Dipartimento di Chimica Biologica, Università di Padova, Istituto di Neuroscienze del CNR, Unità per lo studio delle Biomembrane, Viale G. Colombo 3, 35121 Padūa, Italy.
Free Radic Biol Med. 2004 Oct 1;37(7):1073-80. doi: 10.1016/j.freeradbiomed.2004.06.044.
When rat liver mitochondria are allowed to cycle Ca(2+) and are incubated in the presence of the pro-oxidant menadione, they undergo swelling, membrane potential (DeltaPsi) collapse, and ion release. These effects, which are inhibited by cyclosporin A (CsA), are fully consistent with the opening of the so-called permeability transition pore. However, when Ca(2+) cycling is abolished by EGTA, the mitochondria remain energized (DeltaPsi collapse and swelling are avoided), but Ca(2+) efflux, promoted by the chelating agent, is stimulated by menadione. This stimulation goes together with the release of Mg(2+), K(+), and adenine nucleotides (AdN) and is inhibited by bongkrekic acid (BKA). The effect of menadione is also characterized by biphasic NAD(P)H oxidation which becomes monophasic in the presence of BKA, CsA, or EGTA and by the oxidation of thiol groups not restrained by the above-mentioned inhibitors. These results suggest that BKA acts indirectly by preserving in the matrix a critical amount of AdN without modifying the monophasic oxidation of pyridine nucleotides by menadione. A critical number of thiol groups also seems to be involved in the phenomenon. Their oxidation most probably causes a conformational change on adenine nucleotide translocase with the opening of the "low-conductance state" of the mitochondrial permeability transition, resulting in ion permeability without DeltaPsi disruption and mitochondrial swelling.
当大鼠肝线粒体被允许循环钙离子并在促氧化剂甲萘醌存在的情况下孵育时,它们会发生肿胀、膜电位(ΔΨ)崩溃以及离子释放。这些效应受到环孢素A(CsA)的抑制,与所谓的通透性转换孔的开放完全一致。然而,当用乙二醇双四乙酸(EGTA)消除钙离子循环时,线粒体仍保持能量状态(避免了ΔΨ崩溃和肿胀),但螯合剂促进的钙离子外流受到甲萘醌的刺激。这种刺激伴随着镁离子(Mg²⁺)、钾离子(K⁺)和腺嘌呤核苷酸(AdN)的释放,并受到邦克雷酸(BKA)的抑制。甲萘醌的作用还表现为双相烟酰胺腺嘌呤二核苷酸(磷酸)[NAD(P)H]氧化,在BKA、CsA或EGTA存在时变为单相氧化,以及不受上述抑制剂抑制的巯基氧化。这些结果表明,BKA通过在基质中保留临界量的AdN而间接发挥作用,而不改变甲萘醌对吡啶核苷酸的单相氧化。一定数量的关键巯基似乎也参与了这一现象。它们的氧化很可能导致腺嘌呤核苷酸转位酶的构象变化,伴随着线粒体通透性转换的“低电导状态”的开放,导致离子通透性增加而不破坏ΔΨ和线粒体肿胀。