中文摘要：

目的：研究缺氧/复氧（hypoxia/reoxygenation,H/R）刺激是否可通过激活程序性坏死信号通路诱导心肌细胞坏死。方法 ：体外培养新生大鼠原代心肌细胞,采用缺氧2 h复氧4 h的方法复制心肌细胞损伤模型,碘化丙腚（propidium iodide,PI）染色检测心肌细胞坏死情况,Western blot和免疫共沉淀方法检测程序性坏死信号通路中受体相互作用蛋白1（receptor interacting protein-1,RIP1）、受体相互作用蛋白3（receptor interacting protein-3,RIP3）的蛋白表达水平和RIP1/RIP3复合物Ⅱ形成情况,以及RIP1、RIP3的泛素化水平。结果：与对照组相比,缺氧/复氧组心肌细胞坏死数量明显增多,RIP1、RIP3蛋白表达水平显著升高,RIP1/RIP3复合物Ⅱ的形成增多,且RIP1与RIP3的泛素化水平也有所增加。结论：缺氧复氧可诱导心肌细胞坏死,其机制可能与激活程序性坏死RIP1/RIP3信号通路有关。

英文摘要：

Objective：To study whether hypoxia and reoxygenation（H / R）injury induces the necrosis of cardiomyocyte via regulation of programmed necrosis（necroptosis）. Methods：Neonatal rats ventricular myocytes were isolated from neonatal Sprague –Dawley rats. The incubated cells were subjected to reoxygenation（4 h）after hypoxia（2 h）. The effect of H / R injury on the necrosis of cardiaomyocytes was determined by PI（propidium iodide）staining. Wsetern blot and Co-IP were performed to test the protein expression of receptor interacting protein（RIP）1 and 3,the formation of RIP1 / RIP3 complex Ⅱ,and the ubiquitination level of RIP1 and RIP3. Results：When subjected to reoxygenation（4 h） after hypoxia（2 h）,the protein expression levels of RIP1 and RIP3 were significantly increased compared with those of control. Besides,H / R injury also promoted the formation of RIP1 / RIP3 complex Ⅱand upregulated the ubiquitination of RIP1 and RIP3. Conclusion：Hypoxia and reoxygenation injury could induce the necrosis of cardiomyocytes,which may be involved in regulation of RIP1 / RIP3 dependent programmed necrosis signaling pathway.