摘要
目的检测Wnt-1在大鼠气管上皮损伤修复中的表达及其变化,探讨影响气管干细胞早期增殖分化的分子调控机制。方法氟脲嘧啶(5-FU)作用大鼠离体气管,去除5-FU后分别于0、6、12、24及48 h用Westernblotting及免疫组织化学方法检测气管上皮中Wnt-1的表达。结果5-FU损伤后12 h,气管上皮脱落,仅残留少量G0期细胞表达Wnt-1;去除5-FU后6 h,上皮细胞数目增多,呈扁平状间断分布,Wnt-1表达最强;12 h后,上皮细胞呈立方状,并连接成片,Wnt-1表达逐渐减少;恢复48 h,气管上皮内出现假复层结构,仅见极少量Wnt-1阳性细胞。正常气管上皮Wnt-1表达极少。结论Wnt-1时间依赖性表达与气管上皮损伤修复进程相吻合,说明其在气管干细胞早期增殖分化中起重要调控作用。
Objective To determine the expression of Wnt-1 in rat tracheal epithelium during the regeneration after injury, to explore the mechanisms in the early proliferation and differentiation of the tracheal stem cells. Methods Extracorporeal tracheal injury was induced by 5-FU. Tracheas were taked out on 0, 6, 12, 24, 48 hours after removement of 5-FU. Wnt-1 expression in tracheal epithelium during the process of regeneration was analyzed by immunohistochemistry and Western blotting. Results 1. After treatment with 5-FU for 12 hours, the tracheal epithelium shed and some of the retained cells in GO were Wnt-1 positive. Six hours after the removal of 5-FU, the tracheal rings were covered with flattened epithelial cells. Wnt-1 positive cells increased obviously. At 12 hours after the removal of 5-FU, most of the epithelial cells were cuboidal and merged into pieces, but the Wnt-1 positive cells decreased obviously. At 48 hours, only a few positive cells could be seen with some of the pseudostratified mucociliary epithelium restored similar to its original mode. There were no detectable Wnt-1 positive cells in normal tracheal epithelium. 2.Western blotting analysis showed that there were different Wnt-1 levels at different times after the removal of 5-FU which in accordance with the change of immunohistochemistry. Wnt-1 was minimally detected after treatment with 5-FU for 12 hours, reaching a maximal level at 6 hours after the removal of 5-FU, and then decreased over time. At 48 hours, very low Wnt-1 level was detected. Conclusion The expression of Wnt-1 corresponds with the wound and healing process of tracheal epithelium, suggesting that Wnt-1 may play an important effect in regulating the tracheal stem cell proliferation and differentiation.
出处
《解剖学报》
CAS
CSCD
北大核心
2006年第2期168-171,共4页
Acta Anatomica Sinica
基金
国家自然科学基金资助项目(30170407)
关键词
WNT-1
气管干细胞
增殖
分化
免疫印迹法
大鼠
Wnt-1
Tracheal stem cell
Proliferation
Differentiation
Western blotting
Rat
