【作者】 熊轶；

【导师】 黄中新；

【作者基本信息】 暨南大学 ， 组织学与胚胎学， 2003， 硕士

【摘要】 目的：检测人胎肺成纤维生长因子受体(FGFR)、骨形成蛋白—4(BMP-4)、rasp—21蛋白、甲状腺转录因子—1(TTF-1)和肺表面蛋白—B(SP-B)在Ⅱ型肺泡细胞发育过程中的表达特征，探讨它们在调节Ⅱ型肺泡细胞发育和分化过程中的生物活性作用，以及相互之间的调控意义。 方法：16-35周人胎肺组织，共15例。新鲜组织用4％多聚甲醛灌注，中性甲醛固定48小时以上，常规石蜡包埋，切片4μm；免疫组化SP法检测FGFR、BMP-4、rasp—21蛋白、TTF-1和SP-B在胎肺中的表达特征。 结果：1．胎肺发育早期(12w)，FGFR率先在近端支气管上皮细胞胞质内表达，随着发育，远端呼吸道上皮表达逐渐丰富。18w，近端、远端支气管和肺泡都呈现强阳性，并达到表达高峰。BMP-4在近端和远端的呼吸道上皮表达，呈现强阳性，而肺泡上皮的表达呈现弱阳性。22w至25w，远端呼吸道上皮和肺泡上皮的反应增强。rasp-21蛋白于16w在呼吸道支气管上皮胞质中有较弱表达。18w，不论是近端、远端或肺泡的上皮，表达都较强烈。而到了20w只有近端的反应依然强烈。至发育的末期，这三种因子几乎不表达； 2．TTF-1于16w表达在呼吸道上皮细胞核内，远端呼吸道上皮细胞始终较近端呼吸道上皮阳性细胞丰富。22w，TTF-1逐渐向新形成的呼吸道终末细胞迁移，同时，阳性细胞在间质内呈现成团分布的区域。晚期，呼吸道上皮TTF-1表达较前逐渐减弱，仅集中到终末囊泡和新生肺泡内。18w，SP-B在上皮细胞胞质内开始表达。阳性表达的细胞逐渐地由近端呼吸道上皮往远端呼吸道迁移，强度比早期有所增强，在近端呼吸道上皮中，SP-B阳性细胞逐渐呈现零星分散的状态。35w，SP-B阳性细胞的表达仅定位在肺泡Ⅱ型细胞。 结论：FGFR、BMP-4和rasp21蛋白促使Ⅱ型肺泡细胞增殖分化，使之达到形态上的成熟。TTF-1因子和SP-B促进Ⅱ型肺泡细胞达到功能上的成熟。它们在Ⅱ型肺泡细胞发育的过程中是相互制约、相互影响、相互调控的，Ⅱ型肺泡细胞正常发育有赖于这些细胞因子之间复杂的协调作用。更多还原

【Abstract】 Objective: In this work, distributions of the interrelated proteins: the fibroblast growth factor receptor (FGFR), bone morphogenetic protein-4 (BMP-4), rasp21 protein , thyroid transcription factor-1(TTF-1) and surface protein-b(SP-B) in human fetal lung were determined, their important function properties in regulating lung morphogenesis , differentiation and maturation were studied, as well as their relationships between each other.Methods: 15 specimens of human fetal lungs (from the 12thw to 35thw of gestation) were obtained. The expression of 5 kinds of polypeptides (FGFR, BMP-4, rasp21 protein, TTF-1, and SP-B) were examined by immunohistochemistry and re-stained by SP methods.Results: 1. It is showed that FGFR firstly localized in the airway surface epithelium during early lung (12thw ) development. Its staining in distal bronchiolar epithelial cells is always more intense than that of proximal epithelial cells. Its reaction reached the peak on fetal age 18th w. In this stage, the positive reactions were mainly detected in proximal and distal bronchiolar epithelial cells and also in type II alveolar cells. In 18thw, we can get the staining of BMP-4 in the airway surface epithelium, but its staining is more obvious in airway epithelium than that of type II alveolar cells. From 22nd w to 25th w, the expression of BMP-4 in distal bronchiolar epithelial cells and in type II alveolar cells has reinforced. In 16th w, rasp21 protein were expressed in proximal bronchioles but epithelial cells expressed at low levels. In 18thw, rasp21 protein were expressed throughout fetal lung development and showed at high levers. In 20thw, the intensity of rasp21 protein was expressed weakly except proximal bronchiolar epithelial cells. Furthermore, in the late stage of development, these threefactors all have no active reaction in the fetal lungs. 2. In 16thw, TTF-1 can be detected prominently in the nuclei of distal lung buds. With the development of fetal lungs, the staining of distal epithelial cells reinforced gradually. In the mid of development, TTF-1 located gradually in the newly-formed terminal cell of respiratory tract, meanwhile, several cell masses which located in the mesenchyme can be detected with the antibody of TTF-1. In the late of stage, they were slighter in the fetal lung and only apposition in the type II alveolar cells. The expression of SP-B was later than that of TTF-1 in lung. They were faint in the 18thw lung. With the development, the expression shifted gradually from proximal to distal epithelial cells, and the intensity is stronger than that of early stage. In 35thw, we only detected its expression in the cytoplasm of type II alveolar cells.Conclusion: The positive reactions of FGFR, BMP-4, rasp21 protein, TTF-1 and SP-B were respectively different during different stages, which suggests that their functions are changing at individual developmental periods. FGFR, BMP-4 and rasp21 protein mainly accelerate proliferation and differentiation of the type II alveolar cells, and make they more mature in morphology. TTF-1 and SP-B mainly make it mature in function and suitable in respiration. Normal development of fetal lung depends on these factors mutual coordination.更多还原