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目的复制中子和γ线致肠道损伤的体外模型,探讨ERK通路的变化规律及IL-11对其调控作用,为阐明中子和γ线致伤差异的分子机制并寻找有效的防治措施提供依据。方法采用4 Gy中子和10 Gyγ射线照射IEC-6大鼠肠上皮细胞,并于照射前12 h或照射后即刻给予100 ng/ml的rhIL-11,采用流式细胞术、Western blot和图像分析技术,分别检测IEC-6细胞凋亡和坏死率以及Raf-1、MEK1/2、ERK1/2表达及活化的变化。结果中子和γ线均可造成IEC-6细胞损伤,且前者损伤更重;应用IL-11后,二组凋亡率和坏死率均出现下降。γ射线照射后6~24 h,IEC-6细胞Raf-1表达和活化及MEK1/2活化升高,ERK1/2活化减弱,IL-11处理组于照射后5~15 min可增加Raf-1和MEK1/2活性,照射后6 h抑制ERK1/2活化。中子照后6~24 h,Raf-1表达和活化及MEK1/2活化无明显变化,ERK1/2表达和活化升高,与照射组比较,IL-11处理后6 h,Raf-1表达变化不明显,照射后6~24 h,MEK1/2活化升高,IL-11在照射后6 h抑制ERK1/2活化,照后24 h增加ERK1/2表达及活化。结论中子照射造成IEC-6细胞ERK1/2活化,而γ射线照射则表现为抑制作用;IL-11通过调控ERK通路对二者造成的肠损伤具有防护作用。
OBJECTIVE: To study the changes of ERK pathway and the regulatory role of IL-11 in vitro in order to clarify the molecular mechanism of neutron and γ-ray injury and to find effective prevention and control measures in accordance with. Methods Intestinal epithelial cells of IEC-6 rats were irradiated with 4 Gy neutrons and 10 Gy γ-rays and rhIL-11 (100 ng / ml) was given 12 h before irradiation or immediately after irradiation. Flow cytometry, Western blot and image Analyzed the changes of apoptosis and necrosis of IEC-6 cells and the expression and activation of Raf-1, MEK1 / 2 and ERK1 / 2 respectively. Results Both neutron and γ-ray could cause IEC-6 cell injury, and the former had more severe injury. The apoptosis rate and necrosis rate of both groups decreased after IL-11 administration. The expression of Raf-1 and activation of MEK1 / 2 and the activation of ERK1 / 2 in IEC-6 cells increased from 6 to 24 h after γ-ray irradiation. The activation of ERK1 / 2 in IL-11 treatment group increased Raf-1 And MEK1 / 2 activity, and inhibited ERK1 / 2 activation 6 h after irradiation. At 6-24 h after neutron irradiation, the expression and activation of Raf-1 and the activation of MEK1 / 2 did not change significantly, while the expression and activation of ERK1 / 2 increased. Compared with the irradiated group, the expression of Raf-1 at 6 h after IL- The change of MEK1 / 2 was not obvious at 6-24 h after irradiation. IL-11 inhibited ERK1 / 2 activation 6 h after irradiation, and increased ERK1 / 2 expression and activation 24 h after irradiation. Conclusions Neutron irradiation can induce ERK1 / 2 activation in IEC-6 cells, while γ-ray irradiation can inhibit ERK pathway. IL-11 has a protective effect on intestinal damage induced by both ERK pathways.