Even-Tzur Nurit, Kloog Yoel, Wolf Michael, Elad David
Department of Biomedical Engineering, Faculty of Engineering, Tel Aviv University, Tel Aviv, Israel.
Biophys J. 2008 Sep 15;95(6):2998-3008. doi: 10.1529/biophysj.107.127142. Epub 2008 May 16.
The nasal epithelium is continuously subjected to wall shear stresses (WSS) induced by respiratory airflows. An in vitro experimental model was developed to expose nasal epithelial cells cultured under air-liquid interface conditions to steady airflow-induced WSS. Mucus secretion from epithelial goblet cells was quantified using an enzyme-linked lectinosorbent assay, and modifications of the cytoskeletal structure were qualitatively evaluated from fluorescent stains of actin and beta-tubulin fibers. The results show increased mucus secretion from cells subjected to WSS of 0.1 and 1.0 dyne/cm(2) for more than 15 min in comparison with unstressed cells. The integrity levels of beta-tubulin fibers were significantly lower in cells subjected to WSS than in unstressed cells. The increased mucus secretion in response to WSS was approximately the same in Taxol-free and Taxol-treated cultures, which indicates that there is no direct connection between beta-tubulin fragmentation and mucus secretion. The stressed cells regained their normal cytoskeletal appearance 24 h after the exposure to WSS. The results of this study suggest that WSS have an important role in the mechanical regulation of the nasal surface epithelium function.
鼻上皮持续受到呼吸气流诱导产生的壁面剪应力(WSS)作用。我们建立了一种体外实验模型,使在气液界面条件下培养的鼻上皮细胞暴露于稳定气流诱导的WSS中。使用酶联凝集素吸附测定法定量上皮杯状细胞的黏液分泌,并通过肌动蛋白和β微管蛋白纤维的荧光染色定性评估细胞骨架结构的变化。结果显示,与未受应力的细胞相比,承受0.1和1.0达因/平方厘米WSS超过15分钟的细胞黏液分泌增加。承受WSS的细胞中β微管蛋白纤维的完整性水平显著低于未受应力的细胞。在不含紫杉醇和经紫杉醇处理的培养物中,响应WSS而增加的黏液分泌大致相同,这表明β微管蛋白片段化与黏液分泌之间没有直接联系。受应力的细胞在暴露于WSS 24小时后恢复了正常的细胞骨架外观。本研究结果表明,WSS在鼻表面上皮功能的机械调节中起重要作用。