Zhang Ze-Ying, Wu Yang-Qian, Luo Heng, Liu Dong-Xu
School of Life Sciences, Hubei University Wuhan, Hubei, P. R. China.
Int J Clin Exp Med. 2015 Sep 15;8(9):15535-41. eCollection 2015.
To investigate the relationship between NFAT and C5a/C5aR in C5a/C5aR-mediated kidney Ischemia/reperfusion (I/R) injury, the rats' NRK-52E cell line was used in this study and was distributed into 4 groups, I: the normal control (NC), II: the ischemia/reperfusion (I/R) injury cell model (MG), III: the ischemia/reperfusion (I/R) injury cell model treated with C5a (50 nmol/l) (MG + C5a), IV: the ischemia/reperfusion (I/R) injury cell model treated with C5aR antagonist (2.5 μmol/l) (MG + anti-C5aR). Reverse transcription polymerase chain reaction (RT-PCR), western blot, immunofluorescence and flow cytometry were performed. Nuclear Factor Activated T Cell (NFAT), tumor necrosis factor-α (TNF-α) and interleukin (IL-6) were detected in this study. The results of immunofluorescence showed that NFAT had a nuclear translocation phenomenon during the study. The RT-PCR and WB data indicated that the expression of TNF-α and IL-6 in group III were higher than any other groups. Apoptosis in group III was much serious than other groups. All the results in this study showed that NFAT plays an important role in ischemia/reperfusion injury, it can be induced to up-regulate the inflammatory factor TNF-α and IL-6 by the complement system member C5a/C5aR.
为研究核因子活化T细胞(NFAT)与C5a/C5aR在C5a/C5aR介导的肾脏缺血/再灌注(I/R)损伤中的关系,本研究采用大鼠NRK-52E细胞系,并将其分为4组,I:正常对照组(NC);II:缺血/再灌注(I/R)损伤细胞模型组(MG);III:用C5a(50 nmol/l)处理的缺血/再灌注(I/R)损伤细胞模型组(MG + C5a);IV:用C5aR拮抗剂(2.5 μmol/l)处理的缺血/再灌注(I/R)损伤细胞模型组(MG + anti-C5aR)。进行了逆转录聚合酶链反应(RT-PCR)、蛋白质免疫印迹法、免疫荧光和流式细胞术检测。本研究检测了核因子活化T细胞(NFAT)、肿瘤坏死因子-α(TNF-α)和白细胞介素(IL-6)。免疫荧光结果显示,在研究过程中NFAT存在核转位现象。RT-PCR和蛋白质免疫印迹法数据表明,III组中TNF-α和IL-6的表达高于其他任何组。III组的细胞凋亡比其他组严重得多。本研究的所有结果表明,NFAT在缺血/再灌注损伤中起重要作用,它可被补体系统成员C5a/C5aR诱导上调炎症因子TNF-α和IL-6。