The Mechanism of Mitochondria-Mediated Pathway in the Apoptosis of Platelets in Immune-Induced Bone Marrow Failure

10.4077/CJP.2017.BAF484

Qin Zheng；Yiling Jiang；Aiping Zhang；Lele Cui；Lemin Xia；Meihong Luo

apoptosis ； cyclosporine ； mitochondrial pathway

The Chinese Journal of Physiology

60卷6期（2017 / 12 / 31）

338 - 344

英文

Excessive platelet apoptosis is one of the pathogenic causes of immune-induced bone marrow failure (BMF). The aim of the present study was to explore the role of mitochondria-mediated pathway in the apoptosis of platelets in immune-induced BMF. An immune-induced BMF model was established in mice, which were randomly divided into three groups: normal control (CTL) group, BMF group and cyclosporine (CSA) group (n = 10 in each group). Mice were given 0.027 g/kg CSA daily in the CSA group. Platelet count (PLT), mitochondrial transmembrane potential (ΔΨm), cytochrome C (CytC), phosphatidylserine (PS), calcium ion (Ca^(2+)) and expression of proteins of the mitochondrial apoptotic pathway, including Bak, Bax, caspase-3, caspase-8 and caspase-9, was examined and compared. Compared with the CTL group, the BMF group had significantly a lower level of PLC and ΔΨm, but higher levels of CytC, PS, Ca^(2+) and higher expression levels of Bak, Bax, cleaved caspase-9 and cleaved caspase-3 (P ＜ 0.05). CSA restored the above changes in the BMF model (P ＜ 0.05). Further studies showed that intravenous injection of the caspase-9 inhibitor Z-LE(OMe)HD(OMe)-fluoromethylketone (FMK) into the mice could significantly inhibit apoptosis of the platelets and the effect of CSA treatment when compared to the BMF group, and exerted a better protective effect from apoptosis if the caspase-9 inhibitor was combined with the CSA treatment. These results revealed that platelet apoptosis may play an important role in the reduction of platelet of immune-induced BMF probably through the mitochondrial pathway.

1. Brown, S.B.,Clarke, M.C.,Magowan, L.,Sanderson, H.,Savill, J.(2000).Constitutive death of platelets leading to scavenger receptor-mediated phagocytosis. A caspase-independent cell clearance program.J. Biol. Chem.,275,5987-5996.
2. Chen, J.,Desierto, M.J.,Feng, X.,Biancotto, A.,Young, N.S.(2015).Immune-mediated bone marrow failure in C57BL/6 mice.Exp. Hematol.,43,256-267.
3. Desborough, M.,Hadjinicolaou, A.V.,Chaimani, A.,Trivella, M.,Vyas, P.,Doree, C.,Hopewell, S.,Stanworth, S.J.,Estcourt, L.J.(2016).Alternative agents to prophylactic platelet transfusion for preventing bleeding in people with thrombocytopenia due to chronic bone marrow failure: a meta-analysis and systematic review.Cochrane Database Syst. Rev.,10,CD012055.
4. Jacobson, M.D.,Burne, J.F.,Raff, M.C.(1994).Programmed cell death and Bcl-2 protection in the absence of a nucleus.EMBO J.,13,1899-1910.
5. Jagadish, S.,Rajeev, N.,NaveenKumar, S.K.,Sharath Kumar, K.S.,Paul, M.,Hegde, M.,Basappa, Sadashiva, M.P.,Girish, K.S.,Rangappa, K.S.(2016).Platelet protective efficacy of 3,4,5 trisubstituted isoxazole analogue by inhibiting ROS-mediated apoptosis and platelet aggregation.Mol. Cell. Biochem.,414,137-151.
6. Leytin, V.(2012).Apoptosis in the anucleate platelet.Blood Rev.,26,51-63.
7. Leytin, V.,Allen, D.J.,Mutlu, A.,Gyulkhandanyan, A.V.,Mykhaylov, S.,Freedman, J.(2009).Mitochondrial control of platelet apoptosis: effect of cyclosporin A, an inhibitor of the mitochondrial permeability transition pore.Lab. Invest.,89,374-384.
8. Leytin, V.,Allen, D.J.,Mykhaylov, S.,Lyubimov, E.,Freedman, J.(2006).Thrombin-triggered platelet apoptosis.J. Thromb. Haemost.,4,2656-2663.
9. Leytin, V.,Allen, D.J.,Mykhaylov, S.,Mis, L.,Lyubimov, E.V.,Garvey, B.,Freedman, J.(2004).Pathologic high shear stress induces apoptosis events in human platelets.Biochem. Biophys. Res. Commun.,320,303-310.
10. Leytin, V.,Freedman, J.(2003).Platelet apoptosis in stored platelet concentrates and other models.Transfus. Apher. Sci.,28,285-295.
11. Leytin, V.,Mykhaylov, S.,Starkey, A.F.,Allen, D.J.,Lau, H.,Ni, H.,Semple, J.W.,Lazarus, A.H.,Freedman, J.(2006).Intravenous immunoglobulin inhibits anti-glycoprotein IIb-induced platelet apoptosis in a murine model of immune thrombocytopenia.Brit. J. Haematol.,133,78-82.
12. Li, J.,Xia, Y.,Bertino, A.M.,Coburn, J.P.,Kuter, D.J.(2000).The mechanism of apoptosis in human platelets during storage.Transfusion,40,1320-1329.
13. Liu, H.T.、Zhao, J.M.、Chu, J.X.(2007)。Experimental study of low dose irradiation for treatment of immuno-mediated aplastic anemia in mice。J. Exp. Hematol.，15，510-514。
14. Martin, S.J.,Finucane, D.M.,Amarante-Mendes, G.P.,O'Brien, G.A.,Green, D.R.(1996).Phosphatidylserine externalization during CD95-induced apoptosis of cells and cytoplasts requires ICE/CED-3 protease activity.J. Biol. Chem.,271,28753-28756.
15. Nakao, S.(2016).Diagnostic problems in acquired bone marrow failure syndromes.Int. J. Hematol.,104,151-152.
16. Piguet, P.F.,Vesin, C.(2002).Modulation of platelet caspases and life-span by anti-platelet antibodies in mice.Eur. J. Haematol.,68,253-261.
17. Rand, M.L.,Wang, H.,Bang, K.W.,Poon, K.S.,Packham, M.A.,Freedman, J.(2004).Procoagulant surface exposure and apoptosis in rabbit platelets: association with shortened survival and steadystate senescence.J. Thromb. Haemost.,2,651-659.
18. Sat yamitra, M.,Ney, P.,Graves, J., 3rd,Mullaney, C.,Srinivasan, V.(2012).Mechanism of radioprotection by delta-tocotrienol: pharmacokinetics, pharmacodynamics and modulation of signaling pathways.Brit. J. Radiol.,85,e1093-e1103.
19. Shcherbina, A.,Remold-O'Donnell, E.(1999).Role of caspase in a subset of human platelet activation responses.Blood,93,4222-4231.
20. Vanags, D.M.,Orrenius, S.,Aguilar-Santelises, M.(1997).Alterations in Bcl-2/Bax protein levels in platelets form part of an ionomycin-induced process that resembles apoptosis.Brit. J. Haematol.,99,824-831.
21. Wang, B.,Zheng, J.(2016).Platelet generation in vivo and in vitro.Springerplus,5,787.
22. Wang, Z.,Cai, F.,Chen, X.,Luo, M.,Hu, L.,Lu, Y.(2013).The role of mitochondria-derived reactive oxygen species in hyperthermia-induced platelet apoptosis.PLoS One,8,e75044.
23. Wang, Z.,Wang, J.,Xie, R.,Liu, R.,Lu, Y.(2015).Mitochondriaderived reactive oxygen species play an important role in Doxorubicin-induced platelet apoptosis.Int. J. Mol. Sci.,16,11087-11100.
24. Winkler, J.,Rand, M.L.,Schmugge, M.,Speer, O.(2013).Omi/HtrA2 and XIAP are components of platelet apoptosis signalling.Thromb. Haemost.,109,532-539.
25. Wolf, B.B.,Goldstein, J.C.,Stennicke, H.R.,Beere, H.,Amarante-Mendes, G.P.,Salvesen, G.S.,Green, D.R.(1999).Calpain functions in a caspase-independent manner to promote apoptosis-like events during platelet activation.Blood,94,1683-1692.
26. Yamazaki, H.(2016)。Acquired aplastic anemia。Jpn. J. Clin. Hematol.，57，91-97。