Development of new therapy for airway goblet cell metaplasia by transition to ciliated cells

1.使用高度分化的上皮细胞培养物的体外研究。白细胞介素（IL）-13诱导杯状细胞化生，在哮喘粘液高分泌中起重要作用。在这项研究中，我们问是否消除IL-13可以逆转既定的杯状细胞化生成纤毛上皮。将豚鼠气管原代上皮细胞与含人重组IL-13的培养基在气液界面上培养14天，然后用去除IL-13的培养基继续培养，或在用抗IL-13抗体中和IL-13的条件下培养。IL-13消除2天后，PAS阳性面积以及MUC 5AC蛋白水平迅速下降。4天后，杯状细胞数量显著减少，而纤毛细胞数量则相反地增加。上皮细胞总数没有变化，IL-13消除后5-溴-2 '-脱氧尿苷摄取减少。去除IL-13后，具有纤毛和分泌颗粒的移行细胞增加。用IL-13抗体中和IL-13 5 d，可逆转杯状细胞向纤毛上皮化生，移行细胞也增多.在致敏豚鼠中使用重塑气道的体内研究。用卵蛋白致敏豚鼠气道杯状细胞，观察Th 2抑制剂、抗IL-13抗体和激素对杯状细胞化生的影响。取出气管并用PAS/阿辛蓝染色。通过计算机图像分析评估粘液面积。结果，抑制剂抑制了粘液面积，出现了既有纤毛又有粘液颗粒的移行细胞.抗IL-13治疗可逆转杯状细胞化生为纤毛上皮。杯状细胞向纤毛细胞的转变与此现象有关。抗IL-13治疗可能是哮喘杯状细胞化生的一种治疗策略。

1. In vitro study using highly differentiated epithelial cell culture. Interleukin (IL)-13 induces goblet cell metaplasia and plays an important role in mucus hypersecretion in asthma. In this study, we asked whether elimination of IL-13 could reverse the established goblet cell metaplasia into ciliated epithelia. Primary epithelial cells from guinea pig tracheas were cultured at an air-liquid interface with the medium containing human recombinant IL-13 for 14 days, and continuously cultured with IL-13-eliminated medium, or cultured under the condition of neutralization of IL-13 with anti IL-13 antibody. After 2 days from elimination of IL-13, PAS-positive area as well as MUC5AC protein level rapidly decreased. After 4 days, the number of goblet cells dramatically decreased, while that of ciliated cells inversely increased. Total number of epithelial cells did not change, and 5-bromo-2'-deoxyuridine uptake decreased after IL-13 elimination. Transitional cells with cilia and secretory granules increased after IL-13 elimination. Similarly, the neutralization of IL-13 with anti IL-13 antibody for 5 days reversed the goblet cell metaplasia into ciliated epithelia, and transitional cells also increased.2. In vivo study using remodeled airway in sensitized guinea pigs. Airway goblet cell metaplasia was obtained by repeated ovalbumin sensitization in guinea pigs, and the effects of Th2 inhibitor, anti IL-13 antibody and steroid on goblet cell metaplasia were investigated.,Tracheas were removed and stained with PAS/alcian blue. Mucus area was assessed by computer image analysis. As a result, mucus area was suppressed by the inhibitors, and transitional cells with both cilia and mucus granules appeared.3. Anti IL-13 therapy may reverse goblet cell metaplasia into ciliated epithelia. Transition of goblet cells to ciliated cells is provably involved in this phenomenon. Anti IL-13 therapy may be a therapeutic strategy of established goblet cell metaplasia in asthma.