论文部分内容阅读
目的检测组蛋白去乙酰化酶1(HDAC1)在人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增殖分化的分子调控机制。方法 5-氟尿嘧啶(5-FU)造成人离体气管上皮损伤,应用免疫组织化学和Western blotting方法检测修复过程中各时间点气管上皮HDAC1的表达。结果去除5-FU后0h,气管上皮脱落,仅残留少量G0期细胞,HDAC1表达阴性;3～6h,细胞数量增多,为扁平状,未见明显HDAC1阳性细胞;12h,上皮细胞由扁平变立方并连接成片,HDAC1阳性细胞数明显增多,并可见数个HDAC1阳性细胞围绕着底部1个HDAC1阴性细胞和多个HDAC1阳性细胞间夹着数个HDAC1阴性细胞分布的现象;24h,细胞数继续增多并变复层,HDAC1阳性细胞数最多,累及全层;48h,接近假复层结构,HDAC1阳性细胞数略减少,阳性细胞分布集中于假复层结构顶部即靠近管腔侧;正常气管上皮HDAC1表达阴性。Western blotting分析显示,HDAC1表达量的变化趋势与免疫组织化学结果一致。结论随着气管干细胞分化细胞数增多,HDAC1阳性细胞数明显增多,提示HDAC1可作为气管干细胞由增殖转向分化的分子开关。
Objective To detect the expression of HDAC1 in human tracheal epithelial injury and its changes, and to explore the molecular regulation mechanism that affects the proliferation and differentiation of tracheal stem cells. Methods Human 5-fluorouracil (5-FU) induced injury of human tracheal epithelium in vitro. The expression of HDAC1 in tracheal epithelium was detected by immunohistochemistry and Western blotting. Results After 0-h removal of 5-FU, only a small amount of G0-phase cells remained and the expression of HDAC1 was negative. After 3 to 6 hours, the number of cells was flat and no obvious HDAC1-positive cells were observed. On day 12, epithelial cells were transformed from flattened cubes And connected into a piece, HDAC1 positive cells increased significantly, and can be seen several HDAC1-positive cells around the bottom of a HDAC1-negative cells and a number of HDAC1-positive cells interspersed with several HDAC1-negative cells distribution; 24h, the number of cells continue The number of HDAC1 positive cells increased most and reached the whole layer at 48h. At 48h, the number of HDAC1 positive cells decreased slightly, and the distribution of positive cells concentrated at the top of the pseudostratified structure near the luminal side. In normal tracheal epithelium HDAC1 expression is negative. Western blotting analysis showed that the trend of HDAC1 expression was consistent with the immunohistochemical results. Conclusion With the increase of tracheal stem cell differentiation, the number of HDAC1 positive cells increased obviously, suggesting that HDAC1 can be used as a molecular switch of proliferation and differentiation of tracheal stem cells.