组蛋白去乙酰化酶1在气管干细胞增殖分化过程中的时空表达

发布时间：2018-06-13 23:50

  本文选题：支气管 + 气管干细胞　； 参考：《中国医科大学》2006年硕士论文

【摘要】：组蛋白去乙酰化酶1在人支气管干细胞增殖分化过程中的时空表达 前言 目前,在气管干细胞研究中,我们建立了氟脲嘧啶(Fluorouracil,5-FU)引发离体气管损伤修复模型,论证了气管干细胞位于具有5-FU抗性的G0期细胞中。为阐明气管干细胞增殖分化的调控机制,我们已检测了wnt信号中的wntl,β-catenin,cyclinD1等成员在气管干细胞增殖分化过程中的时空变化情况,证明了Wnt/β-catenin信号传导途径参与调控气管干细胞增殖分化全过程。近来有学者在研究斑马鱼视网膜发育中发现组蛋白去乙酰化酶1(Histone deacetylase 1,HDAC1)促进视网膜干细胞分化成神经元和胶质细胞,提出HDAC1是视网膜干细胞由增殖到分化的开关。那么HDAC1在气管干细胞的增殖分化过程中是否也具有这种开关作用?这在国际上尚无报道。本研究利用免疫组织化学和western blotting方法检测了HDAC1在气管干细胞增殖分化过程中的动态表达情况。 材料和方法 1.制备人支气管损伤模型:在因肺癌肺叶切除组织中取距肺癌组织2cm处支气管(该材料经患者家属及中国医科大学伦理委员会同意)放入无菌袋中在2h内用冰盒运回实验室,无菌PBS反复冲洗,洗净血液和粘液,置于DMEM/F12培养液中(含10%胎牛血清),剪成约2-3mm的气管环,于倒置显微镜下观察纤毛摆动良好,于培养液中加入终浓度为120mg/ml的5-FU,37℃,5%CO_2孵育12h,弃去上述培养液,换成新鲜DMEM/F12液(含10%胎牛血清)继续培养,于换液后0、3、6、12、24、48h分别取出一组织块,10%中性福尔马林固定,石蜡包埋,制成4μm厚的组织切片。另于换液后0、3、6、24、48h取气管组织,解剖显微镜下机械剥离上皮,每个样品约
[Abstract]:The temporal and spatial expression of histone deacetylase 1 in the proliferation and differentiation of human bronchial stem cells. In the study of tracheal stem cells, we established a model of tracheal injury repair induced by Fluorouraciline (5-FU) in vitro, and demonstrated that tracheal stem cells were located in G _ 0 cells with 5-FU resistance. In order to elucidate the regulatory mechanism of the proliferation and differentiation of tracheal stem cells, we have detected the temporal and spatial changes of the members of wnt signal, such as wntland 尾 -cateninine cyclin D1, during the proliferation and differentiation of tracheal stem cells. It is demonstrated that Wnt- 尾 -catenin signaling pathway is involved in regulating the proliferation and differentiation of tracheal stem cells. Histone deacetylase (histone deacetylase 1HDAC1) has been found to promote the differentiation of retinal stem cells into neurons and glial cells in zebrafish retina, suggesting that HDAC1 is the switch from proliferation to differentiation of retinal stem cells. Does HDAC1 have the same switching effect in the proliferation and differentiation of tracheal stem cells? This is not reported internationally. In this study, the dynamic expression of HDAC1 in the proliferation and differentiation of tracheal stem cells was detected by immunohistochemical and western blotting methods. Materials and methods 1. To prepare a human bronchial injury model: the bronchus from the lung cancer tissue 2cm (agreed by the patient's family members and the ethics committee of China Medical University) was removed from the lung lobectomy tissue of lung cancer and placed in an aseptic bag and transported back to the laboratory in an ice box within 2 hours. The aseptic PBS was washed repeatedly, the blood and mucus were washed and placed in the DMEM / F12 culture medium (containing 10% fetal bovine serum), the tracheal rings of about 2-3mm were cut, and the cilia swung well under inverted microscope. After incubation with 5-FU (120mg/ml) at 37 鈩，

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