摘要
目的:活性氧介导的氧化损伤是缺血再灌注损伤的重要机制,本研究通过观察H2O2预处理对氧化损伤的H9c2心肌细胞存活率和细胞凋亡的影响,探讨其保护H9c2心肌细胞的作用机制。方法:体外培养H9c2心肌细胞,取对数生长期细胞用于实验研究。建立H2O2预处理抵抗高浓度H2O2诱导的细胞氧化损伤模型,实验分组如下:(1)正常对照组(CTL);(2)损伤组(INJURY);(3)预处理组+损伤组(PC)。应用CCK8法检测细胞存活率;试剂盒检测胞内MDA水平和T-SOD活性;Hoechst 33258染色观察凋亡形态;Annexin-V/PI双染与流式细胞术检测细胞凋亡率。结果:25μmol/L的H2O2预处理90min能明显地保护H9c2心肌细胞抵抗400μmol/L H2O2诱导的氧化损伤,提高细胞存活率,下调MDA水平,上调SOD活性,抑制细胞凋亡,降低细胞凋亡率。结论:低浓度H2O2预处理能减轻H9c2心肌细胞的氧化损伤,抑制氧化损伤诱导的心肌细胞凋亡,具有很好的抗氧化损伤和抗心肌细胞凋亡的保护作用,其作用机制可能与细胞SOD活性上调有关。H2O2预处理为临床治疗心肌缺血/再灌注损伤提供了一项新策略。
Objective： Oxidative injury mediated by reactive oxygen species was an important mechanism of ischemia- reperfusion injury, The aim of this study is to explore the protection mechanism of H2O2 preconditioning by observing the effects of H2O2 precondi- tioning on H9c2 cells viability and apoptosis induced by oxidative injury. Methods： The H9c2 cells were cultured in vitro. The logarith- mic growth phase cells were used to carry on the research. The cells were divided into three groups as follows： （~ control group （CTL）; ~）injury group （1NGURY）; （~） preconditioning＋injury group （PC）. The cell viability was detected by CCK8. MDA content and T-SOD activities were simultaneously measured. Hoechst33258 staining was used to observe the morphology of apoptotic changes. The percent- age of apoptotic cells was quantified by Annexin V and PI double staining and flow cytometry. Results： It was obvious that the H9c2 cells could be protected from oxidative stress injury induced by 400 txmol/L H2O2 by means of 25 μmol/L H2O2 preconditioning for 90 min, and it could increase cell viability, down-regulate MDA content, up-regulate SOD activity, inhibit apoptosis induced by oxidative injury. Conclusion： Low concentration of H2O2 preconditioning could reduce the oxidative injury of H9c2 cells, and inhibit the H9c2 cells apop- tosis induced by oxidative injury. H2O2 preconditioning could obviously protect the H9c2 cells from oxidative stress injury and reduce apoptosis, and its mechanism may be related to increase SOD activity. The H2O2 pretreatrnent provided a new strategy for clinical treatment of myocardial ischemia/reperfusion injury.
出处
《现代生物医学进展》
CAS
2013年第34期6615-6618,6656,共5页
Progress in Modern Biomedicine
基金
国家自然科学基金面上项目(81170095)
