發(fā)布時(shí)間：2018-06-13 23:50

  本文選題：支氣管 + 氣管干細(xì)胞�。� 參考：《中國醫(yī)科大學(xué)》2006年碩士論文

【摘要】：組蛋白去乙�；�1在人支氣管干細(xì)胞增殖分化過程中的時(shí)空表達(dá) 前言 目前,在氣管干細(xì)胞研究中,我們建立了氟脲嘧啶(Fluorouracil,5-FU)引發(fā)離體氣管損傷修復(fù)模型,論證了氣管干細(xì)胞位于具有5-FU抗性的G0期細(xì)胞中。為闡明氣管干細(xì)胞增殖分化的調(diào)控機(jī)制,我們已檢測了wnt信號中的wntl,β-catenin,cyclinD1等成員在氣管干細(xì)胞增殖分化過程中的時(shí)空變化情況,證明了Wnt/β-catenin信號傳導(dǎo)途徑參與調(diào)控氣管干細(xì)胞增殖分化全過程。近來有學(xué)者在研究斑馬魚視網(wǎng)膜發(fā)育中發(fā)現(xiàn)組蛋白去乙�；�1(Histone deacetylase 1,HDAC1)促進(jìn)視網(wǎng)膜干細(xì)胞分化成神經(jīng)元和膠質(zhì)細(xì)胞,提出HDAC1是視網(wǎng)膜干細(xì)胞由增殖到分化的開關(guān)。那么HDAC1在氣管干細(xì)胞的增殖分化過程中是否也具有這種開關(guān)作用?這在國際上尚無報(bào)道。本研究利用免疫組織化學(xué)和western blotting方法檢測了HDAC1在氣管干細(xì)胞增殖分化過程中的動態(tài)表達(dá)情況。 材料和方法 1.制備人支氣管損傷模型:在因肺癌肺葉切除組織中取距肺癌組織2cm處支氣管(該材料經(jīng)患者家屬及中國醫(yī)科大學(xué)倫理委員會同意)放入無菌袋中在2h內(nèi)用冰盒運(yùn)回實(shí)驗(yàn)室,無菌PBS反復(fù)沖洗,洗凈血液和粘液,置于DMEM/F12培養(yǎng)液中(含10%胎牛血清),剪成約2-3mm的氣管環(huán),于倒置顯微鏡下觀察纖毛擺動良好,于培養(yǎng)液中加入終濃度為120mg/ml的5-FU,37℃,5%CO_2孵育12h,棄去上述培養(yǎng)液,換成新鮮DMEM/F12液(含10%胎牛血清)繼續(xù)培養(yǎng),于換液后0、3、6、12、24、48h分別取出一組織塊,10%中性福爾馬林固定,石蠟包埋,制成4μm厚的組織切片。另于換液后0、3、6、24、48h取氣管組織,解剖顯微鏡下機(jī)械剝離上皮,每個(gè)樣品約
[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，

本文編號：2016051