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

1.高分化上皮细胞培养的体外研究。白介素13诱导杯状细胞化生，在哮喘黏液高分泌中起重要作用。在这项研究中，我们询问消除IL-13是否可以逆转已建立的杯状细胞化生为纤毛上皮。将豚鼠原代气管上皮细胞在含有人重组IL-13的气液界面上培养14天，再用去IL-13的培养液继续培养，或在IL-13与抗IL-13抗体中和的条件下培养。IL-13清除2天后，PAS阳性面积和MUC5AC蛋白水平迅速下降。4天后杯状细胞数量显著减少，纤毛细胞数量则相反增加。清除IL-13后，上皮细胞总数无明显变化，5-溴-2‘-脱氧尿苷摄取减少。IL-13清除后移行细胞纤毛增多，分泌颗粒增多。同样，IL-13与抗IL-13抗体中和5天后，杯状细胞化生为纤毛上皮，移行细胞也增多。致敏豚鼠气道重塑的体内研究。用卵蛋白反复致敏的方法复制豚鼠气道杯状细胞化生模型，观察Th2抑制剂、抗IL-13抗体和类固醇对杯状细胞化生的影响，取气管行PAS/Alcian蓝染色。用计算机图像分析方法测定粘液面积。结果表明，抑制物抑制了粘液面积，出现了纤毛和粘液颗粒兼有的移行细胞。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可能与这一现象有关。抗IL-13治疗可能是哮喘已有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.