多肽pPRDX5tide对心肌低氧损伤保护作用的研究

The protective effect of the peptide pPRDX5tide on myocardial hypoxia injury

目的探讨过氧化物酶5来源的多肽(pPRDX5tide)对心肌细胞低氧损伤的保护作用。方法应用生物信息学对心肌低氧多肽组进行分析,初步筛选在低氧后显著上调的pPRDX5tide为候选生物活性多肽,进一步对其稳定性、保守性等基本信息分析,在细胞水平对H9c2大鼠心肌细胞进行pPRDX5tide的预处理后,分别应用低氧袋和化学模拟物氯化钴(Co Cl2)建立物理和化学性低氧损伤的实验模型,检测细胞上清液乳酸脱氢酶(LDH)的水平;台盼蓝染色法检测细胞死亡率;双氯荧光素(DCFH-DA)染色法检测细胞内活性氧(ROS)水平;蛋白质免疫印迹检测半胱天冬酶-3(Caspase3)、多聚腺苷酸二磷酸核糖聚合酶(PARP)的活化。结果预处理10、20、50μmol/L pPRDX5tide对物理低氧和化学低氧造成的心肌细胞损伤呈剂量依赖性的保护作用。50μmol/L pPRDX5tide能显著降低上清中LDH水平,提高细胞的存活率,降低ROS水平,抑制低氧造成的细胞凋亡。结论 pPRDX5tide能显著抑制低氧对细胞造成的损伤,发挥保护作用。

Objective To explore the protective effects of the peptide(pPRDX5tide)derived from peroxidase 5 on myocardial hypoxia injury.Methods The basic information of pPRDX5tide was analyzed,such as stability,conservatism,by using bioinformatics analysis.After rat myocardial cells H9c2 was pretreated with pPRDX5tide,anaeropacks and cobalt chloride(CoCl 2)was applied to establish physical and chemical hypoxia injury experimenal models.The level of lactate dehydrogenase(LDH)in cell supernatant was tested,the cell death rates was counted through trypan blue staining,the level of reactive oxygen species(ROS)in cells was detected by fluorescein(DCFH-DA)staining and the activation of Caspase 3 and PARP was identified through Western blot.Results Pretreatment with 10,20 and 50μmol/L pPRDX5tide had a dose-dependent protective effect on myocardial cell damage caused by physical hypoxia and chemical hypoxia.50μmol/L pPRDX5tide can significantly reduce the level of LDH in the supernatant,increase the survival rate of cells,decrease the level of ROS,and inhibit apoptosis induced by hypoxia.Conclusion pPRDX5tide can significantly inhibit the damage caused by hypoxia on H9c2 cells.