Blockade of Electron Transport during Ischemia Protects Cardiac Mitochondria

Abstract

Subsarcolemmal mitochondria sustain progressive damage during myocardial ischemia. Ischemia decreases the content of the mitochondrial phospholipid cardiolipin accompanied by a decrease in cytochrome c content and a diminished rate of oxidation through cytochrome oxidase. We propose that during ischemia mitochondria produce reactive oxygen species at sites in the electron transport chain proximal to cytochrome oxidase that contribute to the ischemic damage. Isolated, perfused rabbit hearts were treated with rotenone, an irreversible inhibitor of complex I in the proximal electron transport chain, immediately before ischemia. Rotenone pretreatment preserved the contents of cardiolipin and cytochrome c measured after 45 min of ischemia. The rate of oxidation through cytochrome oxidase also was improved in rotenone-treated hearts. Inhibition of the electron transport chain during ischemia lessens damage to mitochondria. Rotenone treatment of isolated subsarcolemmal mitochondria decreased the production of reactive oxygen species during the oxidation of complex I substrates. Thus, the limitation of electron flow during ischemia preserves cardiolipin content, cytochrome c content, and the rate of oxidation through cytochrome oxidase. The mitochondrial electron transport chain contributes to ischemic mitochondrial damage that in turn augments myocyte injury during subsequent reperfusion. Subsarcolemmal mitochondria sustain progressive damage during myocardial ischemia. Ischemia decreases the content of the mitochondrial phospholipid cardiolipin accompanied by a decrease in cytochrome c content and a diminished rate of oxidation through cytochrome oxidase. We propose that during ischemia mitochondria produce reactive oxygen species at sites in the electron transport chain proximal to cytochrome oxidase that contribute to the ischemic damage. Isolated, perfused rabbit hearts were treated with rotenone, an irreversible inhibitor of complex I in the proximal electron transport chain, immediately before ischemia. Rotenone pretreatment preserved the contents of cardiolipin and cytochrome c measured after 45 min of ischemia. The rate of oxidation through cytochrome oxidase also was improved in rotenone-treated hearts. Inhibition of the electron transport chain during ischemia lessens damage to mitochondria. Rotenone treatment of isolated subsarcolemmal mitochondria decreased the production of reactive oxygen species during the oxidation of complex I substrates. Thus, the limitation of electron flow during ischemia preserves cardiolipin content, cytochrome c content, and the rate of oxidation through cytochrome oxidase. The mitochondrial electron transport chain contributes to ischemic mitochondrial damage that in turn augments myocyte injury during subsequent reperfusion. 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Chem. 2003; Full Text Full Text PDF PubMed Scopus Google leads to of in turn to of cytochrome c and decreased of oxidation through cytochrome oxidase. the of electron flow during ischemia during in ischemia in the heart, the electron transport chain was during ischemia and the content of cytochrome and the rate of oxidation through cytochrome oxidase were measured 45 min of ischemia in the isolated rabbit immediately before ischemia with rotenone, is a and irreversible inhibitor of complex preserved cardiolipin content, cytochrome c content, and oxidation through cytochrome oxidase in of electron transport during ischemia damage to more distal sites in the electron transport isolated of electron transport decreased the production of reactive oxygen species E.J. Moghaddas S. Hoppel C.L. E.J. J. Biol. Chem. 2003; Full Text Full Text PDF PubMed Scopus Google Scholar). Thus, the proximal electron transport chain damage to the distal electron transport chain during the progression of ischemia. damage to mitochondria in turn leads to myocyte damage during reperfusion (2Becker L.B. vanden Hoek T.L. Shao Z.H. Li C.Q. Schumacker P.T. Am. J. Physiol. 1999; 277: H2240-H2246PubMed Google Scholar). were of and were were were The rabbit was used (12Lesnefsky E.J. Tandler B. Ye J. Slabe T.J. Turkaly J. Hoppel C.L. Am. J. Physiol. 1997; 273: H1544-H1554Crossref PubMed Google Scholar). The and of the of the of hearts min of min of with treatment followed by 45 min of ischemia. Rotenone was used to complex I of high complex and irreversible is a high inhibitor of complex of I and III was an in the present and the of oxidative was Rotenone was a of min immediately before ischemia. was at a of The of were the to a of inhibitor in the of the to the used in the oxygen (12Lesnefsky E.J. Tandler B. Ye J. Slabe T.J. Turkaly J. Hoppel C.L. Am. J. Physiol. 1997; 273: H1544-H1554Crossref PubMed Google Scholar). was during inhibitor We that and mitochondrial oxidative were preserved during min of E.J. Slabe T.J. Stoll M.S. Minkler P.E. Hoppel C.L. Am. J. Physiol. 2001; 280: H2770-H2778Crossref PubMed Google Scholar). of mitochondria were isolated (12Lesnefsky E.J. Tandler B. Ye J. Slabe T.J. Turkaly J. Hoppel C.L. Am. J. Physiol. 1997; 273: H1544-H1554Crossref PubMed Google Scholar, 16Palmer J.W. Tandler B. Hoppel C.L. J. Biol. Chem. 1977; 252: 8731-8739Abstract Full Text PDF PubMed Google Scholar, 18Lesnefsky E.J. Slabe T.J. Stoll M.S. Minkler P.E. Hoppel C.L. Am. J. Physiol. 2001; 280: H2770-H2778Crossref PubMed Google Scholar). in mitochondria was measured a oxygen at 30 (12Lesnefsky E.J. Tandler B. Ye J. Slabe T.J. Turkaly J. Hoppel C.L. Am. J. Physiol. 1997; 273: H1544-H1554Crossref PubMed Google Scholar). were the of (12Lesnefsky E.J. Tandler B. Ye J. Slabe T.J. Turkaly J. Hoppel C.L. Am. J. 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Minkler P.E. Hoppel C.L. Biochem. PubMed Scopus Google Scholar). was measured by the of in a E.J. Slabe T.J. Stoll M.S. Minkler P.E. Hoppel C.L. Am. J. Physiol. 2001; 280: H2770-H2778Crossref PubMed Google Scholar, E.J. Stoll M.S. Minkler P.E. Hoppel C.L. Biochem. PubMed Scopus Google Scholar). was in rotenone-treated hearts. heart, the in the the heart, was a of the phospholipid were in The rate of production was measured by the oxidation of the in the of S. Hoppel C. E.J. Arch. Biochem. Biophys. 2003; PubMed Scopus Google Scholar). with in the of to a was measured a hearts were at 30 in a of with production was measured in the of As the of decreased by that the in of are the of the were by of with the of R. J. and of Scholar). of was of rabbit was before ischemia 45 min of ischemia 45 min of ischemia with treatment immediately before ischemia. inhibitor and and flow The of inhibitor decreased pressure to than of the pressure before inhibitor an in pressure decrease in flow pressure 45 min of ischemia was in the of 45 with the of IFM with Rotenone was in with IFM also mitochondrial IFM heart, with and was in and IFM with IFM and As complex III was by in and and rate in myocardial mitochondria isolated rabbit hearts with and ischemia treated of complex I of 45 min ischemia 45 min ischemia through cytochrome oxidase in of rate of 45 min ischemia 45 min ischemia through cytochrome oxidase in of rate of 45 min ischemia rate of in a of and Mitochondria Rotenone in (12Lesnefsky E.J. Tandler B. Ye J. Slabe T.J. Turkaly J. Hoppel C.L. Am. J. Physiol. 1997; 273: H1544-H1554Crossref PubMed Google Scholar), and IFM in ischemic hearts were mitochondria in hearts a of Mitochondria in rabbit hearts treated immediately before ischemia with mitochondria in ischemic hearts The of the inner of the mitochondria were by an in and a of the mitochondria the subsarcolemmal and were in were of and were in in the mitochondrial ischemic hearts of a of of were in a Mitochondria isolated ischemic hearts to ischemic hearts. the of isolated hearts a a of the to the of of inner a that and of a of IFM, was a of that in mitochondria in that were a of by a membrane with the of the interfibrillar mitochondria in more structural than subsarcolemmal mitochondria a that during in and isolated of mitochondria rotenone-treated hearts a of with mitochondria the with Rotenone with through Cytochrome the oxidation of an electron to cytochrome oxidase cytochrome decreased 45 min of ischemia in (12Lesnefsky E.J. Tandler B. Ye J. Slabe T.J. Turkaly J. 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Physiol. 1997; 273: H1544-H1554Crossref PubMed Google Scholar). the decrease in through cytochrome oxidase in SSM, ischemia decrease cytochrome oxidase in with (12Lesnefsky E.J. Tandler B. Ye J. Slabe T.J. Turkaly J. Hoppel C.L. Am. J. Physiol. 1997; 273: H1544-H1554Crossref PubMed Google Scholar). treatment before ischemia decreased cytochrome oxidase in with ischemic and rotenone-treated ischemic hearts the preserved cytochrome oxidase in ischemic hearts treated with is with an of damage to cytochrome oxidase by The decrease in cytochrome oxidase in ischemic hearts treated with that inhibitor damage to cytochrome oxidase by a cytochrome oxidase in min ischemia min treated min treated by of mitochondria by of mitochondria in a of in and IFM and decreased the content of cardiolipin in SSM, whereas the cardiolipin content in IFM E.J. Slabe T.J. Stoll M.S. Minkler P.E. Hoppel C.L. Am. J. Physiol. 2001; 280: H2770-H2778Crossref PubMed Google Scholar). The decrease in cardiolipin content is that the contents of and are preserved during ischemia in E.J. Slabe T.J. Stoll M.S. Minkler P.E. Hoppel C.L. Am. J. Physiol. 2001; 280: H2770-H2778Crossref PubMed Google Scholar). Rotenone treatment preserved the content of cardiolipin in 45 min of ischemia with before ischemia also preserved the content of cardiolipin in content of was by ischemia in IFM ischemia rotenone-treated ischemia was measured in The of was by the of the of IFM, E.J. Slabe T.J. Stoll M.S. Minkler P.E. Hoppel C.L. Am. J. Physiol. 2001; 280: H2770-H2778Crossref PubMed Google Scholar, E.J. Stoll M.S. Minkler P.E. Hoppel C.L. Biochem. PubMed Scopus Google Scholar). Rotenone the of Cytochrome c in during content of cytochrome c decreases at 45 min of ischemia in and in Rotenone treatment before ischemia the decrease in cytochrome c content in before ischemia with also decreased the of cytochrome c to the with Inhibition of electron transport also preserved cytochrome c content in IFM ischemia rotenone-treated and ischemia Rotenone of in III is the of production of reactive oxygen species during ischemia (2Becker L.B. vanden Hoek T.L. Shao Z.H. Li C.Q. Schumacker P.T. Am. J. Physiol. 1999; 277: H2240-H2246PubMed Google Scholar). The complex III augments production during the oxidation of complex I and A. G. 1997; PubMed Scopus Google Scholar, J. J. Biol. 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The with treatment between oxidation through cytochrome oxidase in mitochondria and of the contents of cardiolipin and cytochrome c that treatment with cytochrome oxidase of cardiolipin This was by the of a decrease in cytochrome oxidase in in the of phospholipid in hearts Thus, treatment before ischemia cytochrome oxidase by a ischemia damage to of the that leads to decreased through cytochrome oxidase the preserved contents of cardiolipin and cytochrome c in hearts. The present that ischemic damage to mitochondria is at in by the mitochondria The electron transport chain in the of that cardiolipin during in turn to a decrease in cytochrome c content and decreased oxidation through cytochrome oxidase. damage to mitochondria leads to the production of reactive oxygen species (2Becker L.B. vanden Hoek T.L. Shao Z.H. Li C.Q. Schumacker P.T. Am. J. Physiol. 1999; 277: H2240-H2246PubMed Google Scholar), to the of A. R. J. Mol. Cell. Cardiol. 2003; Full Text Full Text PDF PubMed Scopus Google Scholar), and to the of cytochrome c A. R. J. Mol. Cell. Cardiol. 2003; Full Text Full Text PDF PubMed Scopus Google Scholar) during reperfusion. Cytochrome c A. R. J. Mol. Cell. Cardiol. 2003; Full Text Full Text PDF PubMed Scopus Google Scholar, 2003; PubMed Scopus Google Scholar), to an of in A. R. J. Mol. Cell. Cardiol. 2003; Full Text Full Text PDF PubMed Scopus Google Scholar, 2003; PubMed Scopus Google Scholar). The decrease in cytochrome oxidase in turn production complex III during reperfusion (1Ambrosio G. Zweier J.L. Duilio C. Kuppusamy P. Santoro G. Elia P.P. Tritto I. Cirillo P. Condorelli M. Chiariello M. J. Biol. Chem. 1993; 268: 18532-18541Abstract Full Text PDF PubMed Google Scholar). Thus, is that reperfusion periods of ischemia that damage the distal electron transport chain to damage during reperfusion (1Ambrosio G. Zweier J.L. Duilio C. Kuppusamy P. Santoro G. Elia P.P. Tritto I. Cirillo P. Condorelli M. Chiariello M. J. Biol. Chem. 1993; 268: 18532-18541Abstract Full Text PDF PubMed Google Scholar). mitochondrial injury during ischemia is a that contributes to damage during ischemia and reperfusion. of electron flow during ischemia a and a to mitochondrial damage during in turn the damage that during reperfusion. We J. S. K. and