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背景与目的:上皮生长因子(keratinocyte growth factor,KGF)引起肺Ⅱ型细胞增殖,并保护肺免受外界多种刺激。以往,对慢反应正常肺组织的动力学参数难以测定,然而,近年来发展的技术使得准确测定正常组织细胞动力学指标成为可 能。BrdUrd掺入DNA后,用流式细胞仪分析可测定组织中细胞的增殖状况,本研究的目的是采用上皮生长因子刺激后,探讨鼠肺正常组织细胞增殖动力学改变的情况。方法:采用KGF(5 mg/kg)或生理盐水C_3Hf/Kam小鼠气管内注射,并分别在0,1,2,3,4,5,及7天予以处死,处死前20分钟或6小时,腹腔内给予BrdUrd(60 mg/kg),肺组织切下后,固定于60％酒精,经消化处理形成单个细胞核,在流式细胞仪分析前行BrdUrd及总DNA含量标记染色,计算出标记指数(LI),细胞S期时间(Ts)及倍增时间(T_(pot))等细胞动力学参数;免疫荧光染色技术用于鉴定增殖细胞的类型。结果:(1)建立了上皮生长因子(5 mg/kg)刺激肺Ⅱ型细胞增殖的方法及气管内给药的途径;(2)肺组织LI由正常值0.5％升至给药后第三天的5.5％,第七天回至正常;(3)细胞T_(pot)由75.5天降至4.7天;生理盐水治疗组增殖参数没有变化。结论:上皮生长因子可引起肺Ⅱ型细胞的增殖,主要表现为LI的升高及T_(pot)降低。这种增殖效应短暂,第七天回至正常。这为进一步将KGF作为
BACKGROUND & AIM: Keratinocyte growth factor (KGF) causes proliferation of lung type II cells and protects the lungs from various external stimuli. In the past, the kinetic parameters of slow reacting normal lung tissue have been difficult to determine, however, the techniques developed in recent years make it possible to accurately determine the indices of normal tissue cell kinetics. BrdUrd incorporation into DNA, the proliferation of cells can be measured by flow cytometry analysis of the purpose of this study is the use of epithelial growth factor stimulation to explore mouse lung tissue proliferation kinetics changes. METHODS: KGF (5 mg / kg) or saline C_3Hf / Kam mice were injected intratracheally and were sacrificed at 0, 1, 2, 3, 4, 5 and 7 days, respectively, 20 minutes or 6 hours before sacrifice , BrdUrd (60 mg / kg) was intraperitoneally administered. After the lung tissue was excised, it was fixed in 60% ethanol and digested to form a single nucleus. BrdUrd and total DNA content were stained before flow cytometry analysis, Index (LI), cell S phase (Ts) and doubling time (T pot), were determined. Immunofluorescence staining was used to identify the types of proliferating cells. Results: (1) A method of stimulating the proliferation of pulmonary type Ⅱ cells by epidermal growth factor (5 mg / kg) and the route of intratracheal administration were established. (2) The lung LI increased from 0.5% to the third Days, 5.5% of the days returned to normal on the seventh day; (3) The cell pot decreased from 75.5 days to 4.7 days; and there was no change in the proliferation parameters in the saline treatment group. Conclusion: Epithelial growth factor can induce the proliferation of lung type Ⅱ cells, which mainly includes the increase of LI and the decrease of pot. The proliferative effect was transient and returned to normal on the seventh day. This is for further KGF as