Phosphodiesterase-5 inhibition with sildenafil attenuates cardiomyocyte apoptosis and left ventricular dysfunction in a chronic model of doxorubicin cardiotoxicity
Phosphodiesterase-5 inhibition with sildenafil attenuates cardiomyocyte apoptosis and left ventricular dysfunction in a chronic model of doxorubicin cardiotoxicity
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DOI：
10.1161/01.cir.0000160359.49478.c2
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发表时间：
2005-04-05
期刊：
影响因子：
37.8
通讯作者：
Kukreja, RC
中科院分区：
文献类型：
--
作者：
Fisher, PW;Salloum, F;Kukreja, RC
背景-西地那非，一种磷酸二酯酶-5抑制剂，通过开放线粒体K-ATP通道诱导心肌保护作用，对抗缺血/再灌注损伤。方法和结果-将雄性ICR小鼠随机分配至4种治疗中的1种：生理盐水、西地那非、多柔比星（5 mg/kg IP）和西地那非（0.7 mg/kg IP）加多柔比星（n = 6/组）。使用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记和原位寡核苷酸连接方法评估细胞凋亡。通过免疫荧光测定结蛋白分布。Western blot检测Bcl-2蛋白表达。采用Langendorff模式测量舒张压和率压乘积评价左心功能。还测量了指示阿霉素心脏毒性的ECG变化。对于体外研究，在多柔比星前1小时，将成年心室心肌细胞暴露于多柔比星（1 μ mol/L）、西地那非（1 μ mol/L）（含或不含N-G-硝基-L-精氨酸甲酯（L-NAME）（100 μ mol/L））或5-羟基癸酸酯（100 μ mol/L），并孵育18小时。多柔比星治疗的小鼠表现出增加的细胞凋亡和结蛋白破坏，这在西地那非+多柔比星组中减弱。多柔比星组Bcl-2降低，但西地那非+多柔比星组维持在基础水平。左心室发展压力和率压乘积在阿霉素组中显著降低，但在西地那非+阿霉素组中减弱。ST间期在阿霉素组中显著增加超过8周。在西地那非+多柔比星组中，ST间期相对于基线保持不变。阿霉素在体外引起细胞凋亡、半胱氨酸蛋白酶-3激活和线粒体膜电位破坏显着增加。相比之下，西地那非显着保护阿霉素心脏毒性，但是，这种保护被取消了两个L-NAME和5-hydroxydecanoic.Conclusions -预防性治疗西地那非预防细胞凋亡和左心室功能障碍的慢性模型阿霉素诱导的心肌病。
Background - Sildenafil, a phosphodiesterase-5 inhibitor, induces cardioprotection against ischemia/reperfusion injury via opening of mitochondrial K-ATP channels. It is unclear whether sildenafil would provide similar protection from doxorubicin-induced cardiotoxicity.Methods and Results - Male ICR mice were randomized to 1 of 4 treatments: saline, sildenafil, doxorubicin ( 5 mg/kg IP), and sildenafil (0.7 mg/kg IP) plus doxorubicin ( n = 6 per group). Apoptosis was assessed with the use of terminal deoxynucleotidyl transferase - mediated dUTP nick-end labeling and in situ oligo ligation methods. Desmin distribution was determined via immunofluorescence. Bcl-2 expression was analyzed by Western blot. Left ventricular function was assessed by measuring developed pressure and rate pressure product in Langendorff mode. ECG changes indicative of doxorubicin cardiotoxicity were also measured. For in vitro studies, adult ventricular cardiomyocytes were exposed to doxorubicin ( 1 mu mol/L), sildenafil ( 1 mu mol/L) with or without N-G-nitro-L-arginine methyl ester (L-NAME) ( 100 mu mol/L), or 5-hydroxydecanoate ( 100 mu mol/L) 1 hour before doxorubicin and incubated for 18 hours. Doxorubicin-treated mice demonstrated increased apoptosis and desmin disruption, which was attenuated in the sildenafil + doxorubicin group. Bcl-2 was decreased in the doxorubicin group but was maintained at basal levels in the sildenafil + doxorubicin group. Left ventricular developed pressure and rate pressure product were significantly depressed in the doxorubicin group but were attenuated in the sildenafil + doxorubicin group. ST interval was significantly increased in the doxorubicin group over 8 weeks. In the sildenafil + doxorubicin group, ST interval remained unchanged from baseline. Doxorubicin caused a significant increase in apoptosis, caspase-3 activation, and disruption of mitochondrial membrane potential in vitro. In contrast, sildenafil significantly protected against doxorubicin cardiotoxicity; however, this protection was abolished by both L-NAME and 5-hydroxydecanoate.Conclusions - Prophylactic treatment with sildenafil prevented apoptosis and left ventricular dysfunction in a chronic model of doxorubicin-induced cardiomyopathy.