Cyclosporine A regulate oxidative stress-induced apoptosis in cardiomyocytes: mechanisms via ROS generation, iNOS and Hsp70.
1. Previous study suggested that cyclosporine A (CsA) could partially reduce ischaemia/reperfusion-induced injury in isolated heart, but the mechanism was still unclear. In this study, the possible mechanisms of cyclosporine A in regulating oxidative stress-induced cardiomyocyte apoptosis were examined. 2. Morphological (cell shrinkage, apoptotic body formation, and DNA fragmentation) and biochemical (annexin-V staining for exposed phosphatidylserine residues) evidences showed that both hydrogen peroxide (H(2)O(2)) and hypoxia/reoxygenation could induce apoptotic change in the embryonal rat heart myoblast-derived cells (H9c2). These effects were inhibited by pre-treatment with CsA at concentration of 0.01-1.0 micro M for 24 h, but were increased with 10.0 micro M CsA. 3. While examining the mechanisms of CsA in protecting cardiomyocyte apoptosis, we found that the collapse of mitochondria membrane potential (DeltaPsim) induced by oxidative stress was partially reversed by CsA (0.01-1.0 micro M). 4. Compared to the control, CSA at the concentration of 0.1 and 10.0 micro M significantly increased the level of intracellular reactive oxygen species (ROS) to 117.2+/-12.4% and 234.4+/-9.3%, respectively. Co-incubating with the antioxidant, ascorbic acid (10.0 micro M), could partially reduce the protective effect of CsA (0.01-1.0 micro M) and the toxic effect of 10.0 micro M CsA. 5. Pre-treatment with CsA at concentration of 0.01-1.0 micro M for 24 h produced up-regulation of heat shock protein 70 (Hsp 70), inducible nitric oxide synthase (iNOS) and also induced NO production, indicating that these factors might be associated with the cell protective effects of CsA. 6. These results suggest that CsA could protect the oxidative stress-induced cardiomyocyte apoptosis not only by preventing the loss of DeltaPsim in mitochondria, but also through ROS generation, Hsp70, and iNOS up-regulation.
Chen
Huei-Wen
HW
Department of Medical Research, National Taiwan University Hospital, Taipei, Taiwan.
Chien
Chiang-Ting
CT
Yu
Sung-Liang
SL
Lee
Yuan-Teh
YT
Chen
Wen-Jone
WJ
D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D015687:Cell Hypoxia / Q000502:physiology; D002460:Cell Line; D016572:Cyclosporine / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D018840:HSP70 Heat-Shock Proteins / Q000187:drug effects / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007425:Intracellular Membranes / Q000187:drug effects / Q000502:physiology; D008564:Membrane Potentials / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000187:drug effects / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D018384:Oxidative Stress / Q000502:physiology*; D010100:Oxygen / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism
D018840:HSP70 Heat-Shock Proteins; D017382:Reactive Oxygen Species; D016572:Cyclosporine; D006861:Hydrogen Peroxide; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D010100:Oxygen