Institut National de la Santé et de la Recherche Médicale, U845, Centre de Recherche Croissance and Signalization, 156 Rue de Vaugirard, Paris, France.
J Pharmacol Exp Ther. 2010 Apr;333(1):60-9. doi: 10.1124/jpet.109.162032. Epub 2010 Jan 5.
Two highly potent and selective cystic fibrosis (CF) transmembrane regulator (CFTR) inhibitors have been identified by high-throughput screening: the thiazolidinone CFTR(inh)-172 [3-[(3-trifluoromethyl)phenyl]-5-[(4-carboxyphenyl)methylene]- 2-thioxo-4-thiazolidinone] and the glycine hydrazide GlyH-101 [N-(2-naphthalenyl)-((3,5-dibromo-2,4-dihydroxyphenyl)methylene)glycine hydrazide]. Inhibition of the CFTR chloride channel by these compounds has been suggested to be of pharmacological interest in the treatment of secretory diarrheas and polycystic kidney disease. In addition, functional inhibition of CFTR by CFTR(inh)-172 has been proposed to be sufficient to mimic the CF inflammatory profile. In the present study, we investigated the effects of the two compounds on reactive oxygen species (ROS) production and mitochondrial membrane potential in several cell lines: the CFTR-deficient human lung epithelial IB3-1 (expressing the heterozygous F508del/W1282X mutation), the isogenic CFTR-corrected C38, and HeLa and A549 as non-CFTR-expressing controls. Both inhibitors were able to induce a rapid increase in ROS levels and depolarize mitochondria in the four cell types, suggesting that these effects are independent of CFTR inhibition. In HeLa cells, these events were associated with a decrease in the rate of oxygen consumption, with GlyH-101 demonstrating a higher potency than CFTR(inh)-172. The impact of CFTR inhibitors on inflammatory parameters was also tested in HeLa cells. CFTR(inh)-172, but not GlyH-101, induced nuclear translocation of nuclear factor-kappaB (NF-kappaB). CFTR(inh)-172 slightly decreased interleukin-8 secretion, whereas GlyH-101 induced a slight increase. These results support the conclusion that CFTR inhibitors may exert nonspecific effects regarding ROS production, mitochondrial failure, and activation of the NF-kappaB signaling pathway, independently of CFTR inhibition.
两种高效且选择性的囊性纤维化跨膜调节器 (CFTR) 抑制剂已通过高通量筛选被鉴定出来：噻唑烷二酮 CFTR(inh)-172 [3-[(3-三氟甲基)苯基]-5-[(4-羧基苯基)亚甲基]-2-硫代-4-噻唑啉酮]和甘氨酸酰肼 GlyH-101 [N-(2-萘基)-((3,5-二溴-2,4-二羟基苯基)亚甲基)甘氨酸酰肼]。这些化合物对 CFTR 氯离子通道的抑制作用被认为在治疗分泌性腹泻和多囊肾病方面具有药理学意义。此外，CFTR(inh)-172 对 CFTR 的功能抑制已被提出足以模拟 CF 的炎症特征。在本研究中，我们研究了这两种化合物对几种细胞系中活性氧 (ROS) 产生和线粒体膜电位的影响：CFTR 缺陷的人肺上皮细胞 IB3-1（表达杂合 F508del/W1282X 突变）、同源 CFTR 校正的 C38，以及 HeLa 和 A549 作为非 CFTR 表达对照。两种抑制剂都能够快速诱导 ROS 水平升高并使四种细胞类型中的线粒体去极化，表明这些作用独立于 CFTR 抑制。在 HeLa 细胞中，这些事件与耗氧量的降低有关，GlyH-101 的作用比 CFTR(inh)-172 更强。还在 HeLa 细胞中测试了 CFTR 抑制剂对炎症参数的影响。CFTR(inh)-172 但不是 GlyH-101 诱导核因子-κB (NF-κB) 的核易位。CFTR(inh)-172 轻度降低白细胞介素-8 的分泌，而 GlyH-101 则诱导轻度增加。这些结果支持 CFTR 抑制剂可能对 ROS 产生、线粒体衰竭和 NF-κB 信号通路的激活产生非特异性影响的结论，而与 CFTR 抑制无关。
