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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 observe the effect of necrostatin-1st variant (Nec-1s) on myocardial ischemia-reperfusion injury induced by programmed cell necrosis in neonatal rats. Methods: Adult male FVB / N mice were randomly divided into ischemia / reperfusion (I / R + Buffer) group, Nec-1s low dose group (Nec-1sL group), Nec-1s normal dose group -1sN group) and Nec-1s high-dose group (Nec-1sH group). The model of myocardial ischemia was established by blocking the left anterior descending coronary artery (coronary artery). The mice were injected intraperitoneally with different concentrations of Nec-1s or corresponding concentrations of DMSO as controls at 30 min before ligation. Myocardial infarction area was observed 24h after myocardial reperfusion, recovery of cardiac function was observed by Langendroff system in vitro, and necrosis of cardiomyocytes was observed by PI staining and LDH level. Western blotting was used to observe the expression of RIP1 / RIP3 / MLKL expression changes. Results: Compared with I / R + Buffer group, myocardial infarct size was significantly decreased in Nec-1sN and Nec-1sH groups, cardiac function was significantly restored during myocardial reperfusion, LDH release was reduced and the number of PI-positive cardiomyocytes was decreased. Western blotting results Showed decreased expression of the cytopathic necrotic pathway protein RIP1 / RIP3 / MLKL. Nec-1sL group related indicators although the change, but no statistically significant difference. CONCLUSION: Nec-1s preconditioning can relieve myocardial ischemia-reperfusion injury by inhibiting RIP1 kinase activity and attenuating cardiomyocyte necrosis in vivo and in vitro.