In vitro pharmacologic restoration of CFTR-mediated chloride transport with sodium 4-phenylbutyrate in cystic fibrosis epithelial cells containing Delta F508-CFTR
In vitro pharmacologic restoration of CFTR-mediated chloride transport with sodium 4-phenylbutyrate in cystic fibrosis epithelial cells containing Delta F508-CFTR
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DOI：
10.1172/jci119788
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发表时间：
1997-11-15
影响因子：
15.9
通讯作者：
Zeitlin, PL
中科院分区：
文献类型：
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作者：
Rubenstein, RC;Egan, ME;Zeitlin, PL
最常见的囊性纤维化跨膜传导调节因子突变Delta F508-CFTR是一个部分功能的氯通道，保留在内质网中并被降解。我们假设，已知的转录调节因子4-苯丁酸钠(4PBA)将使更大比例的Delta F508-CFTR逃脱降解并出现在细胞表面，来自CF患者的鼻息肉上皮原代培养(Delta F508纯合或杂合)，或CF支气管上皮细胞系IB3-1(Delta F508/W1282X)在培养中暴露于4PBA长达7d。4浓度为0.1和2 mM的PBA可使Forsklin激活的氯离子分泌功能恢复。用0.1-1 mM的4PBA和5 mM的4PBA处理IB3-1细胞和5 mM的原代鼻黏膜上皮细胞，均可出现与高尔基体低聚糖的添加和修饰相一致的高分子形式的CFTR。全细胞裂解物免疫印迹法检测到，4PBA处理的CF3-1细胞的免疫细胞化学与Delta F508-CFTR量的增加一致。这些数据表明，4PBA是一种有希望的药理药物，可用于纠正携带Delta F508突变的CF患者的CF表型。
The most common cystic fibrosis transmembrane conductance regulator mutation, Delta F508-CFTR, is a partially functional chloride channel that is retained in the endoplasmic reticulum and degraded. We hypothesize that a known transcriptional regulator, sodium 4-phenylbutyrate (4PBA), will enable a greater fraction of Delta F508-CFTR to escape degradation and appear at the cell surface, Primary cultures of nasal polyp epithelia from CF patients (Delta F508 homozygous or heterozygous), or the CF bronchial epithelial cell line IB3-1 (Delta F508/W1282X) were exposed to 4PBA for up to 7 d in culture. 4PBA treatment at concentrations of 0.1 and 2 mM resulted in the restoration of forskolin-activated chloride secretion. Protein kinase A-activated, linear, 10 pS chloride channels appeared at the plasma membrane of IB3-1 cells at the tested concentration of 2.5 mM, Treatment of IB3-1 cells with 0.1-1 mM 4PBA and primary nasal epithelia with 5 mM 4PBA also resulted in the appearance of higher molecular mass forms of CFTR consistent with addition and modification of oligosaccharides in the Golgi apparatus, as detected by immunoblotting of whole cell lysates with anti-CFTR antisera, Immunocytochemistry in CF epithelial cells treated with 4PBA was consistent with increasing amounts of Delta F508-CFTR. These data indicate that 4PBA is a promising pharmacologic agent for inducing correction of the CF phenotype in CF patients carrying the Delta F508 mutation.