Ceramide in the antiapoptotic effect of ischemic preconditioning
Ceramide in the antiapoptotic effect of ischemic preconditioning
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DOI：
10.1152/ajpheart.00638.2003
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发表时间：
2004-01-01
影响因子：
4.8
通讯作者：
Ovize, M
中科院分区：
文献类型：
--
作者：
Argaud, L;Prigent, AF;Ovize, M
虽然缺血预处理（PC）抑制缺血再灌注心肌细胞凋亡的机制尚不清楚，但有证据表明第二信使神经酰胺的作用。我们在体内研究PC是否可能影响神经酰胺和sn-1，2-二酰基甘油（DAG）的生产，并减弱缺血期间的细胞凋亡。家兔缺血30 min，再灌注4 h。在此之前，他们接受任何干预（对照组）或一次5分钟的缺血，随后5分钟的再灌注（PC组），或鞘磷脂酶抑制剂D 609的静脉给药。在实验的不同时间点，使用DAG激酶测定法测定心肌神经酰胺和DAG含量。通过夹心酶免疫测定法检测和定量细胞凋亡。AR和梗死面积均采用蓝色染料注射和氯化三苯基四氮唑染色进行测量。对照心脏在长时间缺血5分钟时表现出神经酰胺产生的峰值，平均值为64 +/-5 ng/mg组织（P < 0.05，基线时为48 +/-4 ng/mg）。相反，缺血PC和D 609防止神经酰胺增加在长期缺血。缺血30分钟时，PC心脏的心肌DAG含量仅增加。与对照组相比，预处理组和D 609组的细胞凋亡较少，梗死面积有限。这些结果表明，PC的抗凋亡作用可能是由于减少神经酰胺的生产过程中持续缺血在兔心脏。
Although the mechanism by which ischemic preconditioning (PC) inhibits myocardial apoptosis during ischemia-reperfusion is unclear, evidence indicates a role for the secondary messenger ceramide. We investigated in vivo whether PC may affect ceramide and sn-1,2-diacylglycerol (DAG) production, and attenuate apoptosis during ischemia. Rabbits underwent 30 min of ischemia, followed by 4 h of reperfusion. Before this, they received either no intervention (control group) or one episode of 5 min of ischemia, followed by 5 min of reperfusion (PC group), or an intravenous administration of the sphingomyelinase inhibitor D609. Myocardial content of ceramide and DAG was measured using the DAG kinase assay at different time points of the experiment. Apoptosis was detected and quantified by a sandwich enzyme immunoassay. Both AR and infarct size were measured using blue dye injection and triphenyltetrazolium chloride staining. Control hearts exhibited a peak of ceramide production at 5 min of the prolonged ischemia, with a mean value averaging 64 +/- 5 ng/mg tissue (P < 0.05 vs. 48 +/- 4 ng/mg at baseline). In contrast, ischemic PC and D609 prevented ceramide increase during the prolonged ischemia. Myocardial DAG content was increased only in PC hearts at 30 min of ischemia. Preconditioned and D609 groups developed less apoptosis, as well as a limited infarct size, compared with the control group. These results suggest that the antiapoptotic effect of PC may be due to a reduced ceramide production during sustained ischemia in the rabbit heart.