Unfractionated heparin reduces the elasticity of sputum from patients with cystic fibrosis
Unfractionated heparin reduces the elasticity of sputum from patients with cystic fibrosis
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DOI：
10.1152/ajplung.00206.2007
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发表时间：
2007-11-01
影响因子：
4.9
通讯作者：
Shute, Janis K.
中科院分区：
文献类型：
--
作者：
Broughton-Head, Victoria J.;Shur, Jagdeep;Shute, Janis K.
囊性纤维化（CF）患者的气道粘液阻塞降低了肺功能，引起感染，并限制了包括基因治疗载体在内的吸入药物向靶细胞的递送。并非所有患者对目前可用的粘液溶解剂都有反应，需要新的方法。我们的目的是研究普通肝素（UFH）对痰液形态学和流变学的体外影响，以及UFH对成人CF患者痰液中200 nm纳米球扩散的影响。共聚焦激光扫描显微镜被用来成像荧光染色的肌动蛋白和CF痰的DNA成分，和原子力显微镜被用来成像分离的DNA网络。采用动态振荡流变仪测定CF痰液的粘弹性。使用基于Boyden室的测定通过CF痰测量纳米球扩散。CF痰液中的肌动蛋白- DNA束被浓度为0.1-10 mg/ml的UFH分解，并且UFH增强了来自接受α-脱氧核糖核酸酶治疗的患者的痰液中的核酸内切酶活性。UFH显著降低了未接受α-羟链核酸酶治疗患者CF痰液的弹性和屈服应力，但未降低其粘度。肝素剂量依赖性地显著增加了纳米球通过未接受α-链脲醛酶治疗患者CF痰液的扩散，从基线时的10.5 +/- 2.5%增加至10 mg/ml时的36.9 +/- 4.4%，但在接受α-链脲醛酶治疗的患者中更有效，在0.1 mg/ml时具有最大效应。因此，UFH的粘膜活性特性表明其作为囊性纤维化患者的新治疗方法的潜力。
Mucus obstruction of the airway in patients with cystic fibrosis (CF) reduces lung function, invites infection, and limits delivery of inhaled drugs including gene therapy vectors to target cells. Not all patients respond to presently available mucolytics, and new approaches are needed. Our objectives were to investigate the in vitro effects of unfractionated heparin (UFH) on the morphology and rheology of sputum and the effect of UFH on diffusion of 200-nm nanospheres through sputum from adult CF patients. Confocal laser scanning microscopy was used to image fluorescently stained actin and DNA components of CF sputum, and atomic force microscopy was used to image isolated DNA networks. The viscoelasticity of CF sputum was measured using dynamic oscillatory rheometry. Nanosphere diffusion was measured through CF sputum using a Boyden chamber- based assay. Actin- DNA bundles in CF sputum were disaggregated by UFH at concentrations of 0.1-10 mg/ml, and UFH enhanced the endonuclease activity in sputum from patients on dornase alfa therapy. UFH significantly reduced the elasticity and yield stress, but not the viscosity, of CF sputum from patients not receiving dornase alfa therapy. Heparin dose- dependently significantly increased the diffusion of nanospheres through CF sputum from patients not on dornase alfa therapy from 10.5 +/- 2.5% at baseline to 36.9 +/- 4.4% at 10 mg/ml but was more potent, with maximal effect at 0.1 mg/ml, in patients who were on dornase alfa therapy. Thus the mucoactive properties of UFH indicate its potential as a new therapeutic approach in patients with cystic fibrosis.