Necrostatin-1 stablevariant通过抑制细胞程序性坏死减轻心肌缺血再灌注损伤的实验研究  

Necrostatin-1 stable variant suppress necroptosis in ischemic-reperfusion hearts

作　　者：秦东泽[1] 张涌[1] 郭林静[1] 梁法禹[1] 韩清华[2] 

机构地区：[1]山西医科大学第一医院心胸外科,太原030001 [2]山西医科大学第一医院心内科

出　　处：《临床心血管病杂志》2017年第5期455-460,共6页Journal of Clinical Cardiology

基　　金：山西省科技发展计划(社会发展)项目(No:20130313016-12)

摘　　要：目的:观察necrostatin-1stable variant(Nec-1s)抑制心肌细胞程序性坏死对小鼠心肌缺血再灌注损伤的影响。方法:成年雄性FVB/N小鼠被随机分为缺血再灌注组(I/R+Buffer组)、Nec-1s低剂量处理组(Nec-1sL组)、Nec-1s正常剂量处理组(Nec-1sN组)和Nec-1s高剂量处理组(Nec-1sH组)。通过阻断小鼠冠状动脉(冠脉)左前降支建立心肌缺血模型,并于结扎前30min给予小鼠腹腔注射不同浓度Nec-1s或相应浓度的DMSO作为对照。在心肌再灌注24h后,观察不同组间心肌梗死面积的改变,离体Langendroff系统观察心功能的恢复,通过PI染色法和LDH水平观察心肌细胞的坏死,Western blotting观察程序性坏死相关蛋白RIP1/RIP3/MLKL的表达变化。结果:与I/R+Buffer组比较,Nec-1sN组和Nec-1sH组心肌梗死面积明显减少,心肌再灌注期心功能明显恢复,LDH释放降低,PI染色阳性心肌细胞数目减少;Western blotting结果显示,细胞程序性坏死通路蛋白RIP1/RIP3/MLKL表达降低。Nec-1sL组相关指标虽有改变,但无明显统计学差异。结论:Nec-1s预处理通过抑制RIP1激酶活性,减轻在体和离体缺血再灌注小鼠心肌细胞坏死,从而减轻心肌缺血再灌注损伤。Objective：To investigate the effects of necrostatin-1 stable variant on myocardial necroptosis in experimental mice with ischemical reperfusion injury. Method： Male FVB/N mice were divided randomly into 4 groups.. I/R plus DMSO buffer group （n=8）, I/R plus low dose Nec-ls group （0. 6 mg/kg, n=8）, I/R plus normal dose Nec-ls group （3 mg/kg, n=8） and I/R plus high dose Nec-ls group （6 mg/kg, n=8）. The hearts were performed occlusion of the left anterior descending coronary artery 30 min followed by 24 h reperfusion and intraperitional injection with Nec-ls or 0. 002% DMSO as control in I/R plus DMSO group 30 min before operation. The myocardial infarction size were exam- ined; protein expressions of RIP1, RIP3 and MLKL were determined by Western blotting analysis. The mice were pre- pared by Langendroff method through same grouping method. The myocardial necrosis were detected by PI staining and LDH level in the perfusion effluent. Result： Compared with I/R plus Buffer group, the Nec-ls N group and Nec-ls H group presented decreased infarction size, LDH level and PI positive cells, improved cardiac function, down-regulated protein level of RIP1, RIP3 and MLKL. Conclusion： Nec-ls may inhibite RIPK1 and reduce myocardial necroptosis, thereby attenuate I/R injury in mice heart.

关 键 词：心肌缺血再灌注损伤 Necrostatin-1stable VARIANT 细胞程序性坏死 心脏功能 

分 类 号：R542.2[医药卫生—心血管疾病]