摘要
目的观察离体大鼠气管上皮损伤修复过程中气管干细胞的动态变化.方法用氟脲嘧啶（5-FU）造成离体大鼠气管上皮损伤,应用间接免疫荧光法和Western blotting动态观测了修复过程中各时间点气管上皮ABCG2的表达.结果 1.5-FU作用12 h后大部分气管上皮脱落,残留的G0期细胞中部分可见ABCG2表达阳性.去除5-FU后3 h细胞数目增多,为扁平状,ABCG2阳性细胞数也随之增多;6～9 h上皮细胞由扁平变为立方,ABCG2阳性细胞数继续增多;24 h上皮细胞呈立方状,并连接成片,ABCG2阳性细胞较前明显减少;48 h气管上皮接近恢复假复层结构,此时仅见极少量ABCG2阳性细胞.正常气管上皮未检测到ABCG2表达.2.Western blotting分析表明,ABCG2表达量的变化趋势与免疫荧光结果一致.结论 ABCG2的表达与干细胞的数量呈正相关,可作为气管干细胞的标志物.
Objective To explore the dynamic changes of tracheal stem cells during tracheal regeneration after injury induced by fluorouracil （5-FU） in rats. Methods Extracorporeal tracheal injury （Wistar rats） was induced by 5-FU. ABCG2 expression in tracheal epithelium during the process of regeneration was analyzed by indirect immunofluorescence and Western blotting. Results 1 .After treatment with 5-FU for 12 hours, the tracheal epithelium shed and there were ABCG2 positive cells among residual cells in G0. Three hours after the removal of 5-FU, the tracheal rings were covered with flattened epithelial cells. ABCG2 positive cells increased slightly. Six-nine hours after the removal of 5-FU, the epithelial cells changed into cuboidal, correspondingly, the ABCG2 positive cells increased obviously; At 24 hours after the removal of 5-FU, most of the epithelial cells were cuboidal and merged into pieces, but the ABCG2 positive cells decreased obviously; Until 48 hours after the removal of 5- FU,only a few positive ceils could be seen with the pseudostratified mucociliary epithelium restored similar to its original mode. There were no detectable ABCG2 positive cells in normal tracheal epithelium. 2. Western blotting analysis showed that there were different ABCG2 levels at different times after the removal of 5-FU which in accordance with the. change of immunofluorescene. ABCG2 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. Until 48 hours after the removal of 5-FU, very low ABCG2 level was detected. Conclusion The expression of ABCG2 is correlated to the number of tracheal stem cells, suggesting that ABCG2 may serve as a marker for isolating stem cells from tracheal epithelium.
出处
《解剖学报》
CAS
CSCD
北大核心
2006年第2期163-167,共5页
Acta Anatomica Sinica
基金
国家自然科学基金资助项目(30170407)
