Schophuizen Carolien M S, Hoenderop Joost G J, Masereeuw Rosalinde, Heuvel Lambert P van den
Department of Pediatric Nephrology, Radboudumc 6525 GA Nijmegen, The Netherlands.
Department of Physiology, Radboud Institute for Molecular Life Sciences, Radboudumc 6525 GA Nijmegen, The Netherlands.
Cells. 2015 Jun 26;4(3):234-52. doi: 10.3390/cells4030234.
In renal failure, the systemic accumulation of uremic waste products is strongly associated with the development of a chronic inflammatory state. Here, the effect of cationic uremic toxins on the release of inflammatory cytokines and endothelin-1 (ET-1) was investigated in conditionally immortalized proximal tubule epithelial cells (ciPTEC). Additionally, we examined the effects of ET-1 on the cellular uptake mediated by organic cation transporters (OCTs). Exposure of ciPTEC to cationic uremic toxins initiated production of the inflammatory cytokines IL-6 (117 ± 3%, p < 0.001), IL-8 (122 ± 3%, p < 0.001), and ET-1 (134 ± 5%, p < 0.001). This was accompanied by a down-regulation of OCT mediated 4-(4-(dimethylamino)styryl)-N-methylpyridinium-iodide (ASP+) uptake in ciPTEC at 30 min (23 ± 4%, p < 0.001), which restored within 60 min of incubation. Exposure to ET-1 for 24 h increased the ASP+ uptake significantly (20 ± 5%, p < 0.001). These effects could be blocked by BQ-788, indicating activation of an ET-B-receptor-mediated signaling pathway. Downstream the receptor, iNOS inhibition by (N(G)-monomethyl-l-arginine) l-NMMA acetate or aminoguanidine, as well as protein kinase C activation, ameliorated the short-term effects. These results indicate that uremia results in the release of cytokines and ET-1 from human proximal tubule cells, in vitro. Furthermore, ET-1 exposure was found to regulate proximal tubular OCT transport activity in a differential, time-dependent, fashion.
在肾衰竭中,尿毒症废物产物的全身蓄积与慢性炎症状态的发展密切相关。在此,我们研究了阳离子尿毒症毒素对条件永生化近端肾小管上皮细胞(ciPTEC)中炎性细胞因子和内皮素-1(ET-1)释放的影响。此外,我们还检测了ET-1对有机阳离子转运体(OCTs)介导的细胞摄取的影响。将ciPTEC暴露于阳离子尿毒症毒素会引发炎性细胞因子IL-6(117±3%,p<0.001)、IL-8(122±3%,p<0.001)和ET-1(134±5%,p<0.001)的产生。这伴随着ciPTEC中OCT介导的4-(4-(二甲氨基)苯乙烯基)-N-甲基吡啶碘化物(ASP+)摄取在30分钟时下调(23±4%,p<0.001),而在孵育60分钟内恢复。暴露于ET-1 24小时会显著增加ASP+摄取(20±5%,p<0.001)。这些效应可被BQ-788阻断,表明激活了ET-B受体介导的信号通路。在受体下游,用(N(G)-单甲基-L-精氨酸)L-NMMA乙酸盐或氨基胍抑制诱导型一氧化氮合酶(iNOS)以及蛋白激酶C激活可改善短期效应。这些结果表明,尿毒症在体外会导致人近端肾小管细胞释放细胞因子和ET-1。此外,发现ET-1暴露以不同的、时间依赖性的方式调节近端肾小管OCT转运活性。