Song Hyunkeun, Han Il-Yong, Kim Yeonye, Kim Young Hwan, Choi Il-Whan, Seo Su-Kil, Jung So Young, Park SaeGwang, Kang Mi Seon
Departmentof Microbiology and Immunology, College of Medicine, INJE University, Bockjiro 75, BusanjinGu, Busan 614-735, Republic of Korea.
Department of Thoracic and Cardiovascular Surgery, College of Medicine, INJE University, Bockjiro 75, BusanjinGu, Busan 614-735, Republic of Korea.
Life Sci. 2015 Apr 1;126:69-75. doi: 10.1016/j.lfs.2015.02.004. Epub 2015 Mar 2.
Ischemia/reperfusion injury (IRI), resulting from hypoxic damage within a graft, is the leading cause of cell death and graft rejection. In this study, we investigated whether a HIF-1α inhibitor or various antioxidants were able to prevent ischemic injury in a cellular model in which experimental hypoxia was induced using CoCl2.
The ischemic injury induced in HK-2 cells by CoCl2 was validated by increased reactive oxygen species (ROS) production, reduced cell viability, and increased apoptosis at different times and doses. The preventative effects of various anti-oxidants on ischemic injury were evaluated using ROS levels, cell viability, and apoptosis. The MAPK phosphorylation status and Bcl2/Bax expression levels were evaluated after treatment with various antioxidants.
The increase in ROS induced by hypoxia was significantly inhibited by NAC and CAPE, but not by any other treatment. The reduction in cell viability induced by CoCl2 was significantly inhibited by NAC and DPI, but not by any other treatment. The apoptosis induced by CoCl2 was also significantly inhibited by NAC and DPI, but not by any other treatment. Moreover, NAC and DPI prevented CoCl2-induced apoptosis in HK-2 cells in a dose- and time-dependent manner. Treatment of CoCl2 and HK-2 cells treated with DPI, but not NAC, significantly induced ERK activation and Bcl2 expression. NAC and DPI treatment prevented the apoptosis of cells cultured under hypoxic conditions.
Our results suggest that DPI should be investigated further as a novel protective agent that prevents kidney ischemia.
移植器官内的缺氧损伤导致的缺血/再灌注损伤（IRI）是细胞死亡和移植排斥的主要原因。在本研究中，我们调查了缺氧诱导因子-1α（HIF-1α）抑制剂或各种抗氧化剂是否能够在使用氯化钴诱导实验性缺氧的细胞模型中预防缺血性损伤。
通过在不同时间和剂量下活性氧（ROS）生成增加、细胞活力降低和细胞凋亡增加，验证氯化钴在HK-2细胞中诱导的缺血性损伤。使用ROS水平、细胞活力和细胞凋亡评估各种抗氧化剂对缺血性损伤的预防作用。在用各种抗氧化剂处理后，评估丝裂原活化蛋白激酶（MAPK）的磷酸化状态和Bcl-2/Bax表达水平。
N-乙酰半胱氨酸（NAC）和咖啡酸苯乙酯（CAPE）可显著抑制缺氧诱导的ROS增加，但其他处理无此效果。NAC和二苯基碘鎓（DPI）可显著抑制氯化钴诱导的细胞活力降低，但其他处理无此效果。氯化钴诱导的细胞凋亡也被NAC和DPI显著抑制，但其他处理无此效果。此外，NAC和DPI以剂量和时间依赖性方式预防氯化钴诱导的HK-2细胞凋亡。用DPI而非NAC处理氯化钴和HK-2细胞可显著诱导细胞外信号调节激酶（ERK）激活和Bcl-2表达。NAC和DPI处理可预防缺氧条件下培养的细胞凋亡。
我们的结果表明，DPI作为一种预防肾脏缺血的新型保护剂应进一步研究。
