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目的探索胰岛素样生长因子结合蛋白(IGFBP)2在新生鼠肺发育中的作用。方法SD孕鼠80只分为4组,即A:对照组;B:地塞米松(Dex)1组;C:Dex2组;D:维A酸(RA)组。A、B、D组分别于孕18～20d皮下注射生理盐水、Dex,腹腔注射RA;C组于生后1～3d皮下注射Dex。各组分别于孕18、20、21d(C组孕期不取)、生后1、3、5、7、10、14、21d取肺标本作形态学检查、免疫组织化学、Westernblot及RT PCR检测。结果1.肺组织形态学:B、C组早期肺泡发育提前,数目多,壁薄,晚期则明显落后于A、D组。2.IGFBP2肺表达:A组中IGFBP2主要在胎肺组织中表达,孕18d左右表达最强,随后表达渐减弱。B、C组表达趋势同A组,但生后各时间点均较A组增强;D组生后各时间点均较A组减弱。3.Westernblot:IGFBP2多肽表达强度各组孕18d表达最强,随后渐降低;B、C组生后各时间点表达强度明显高于A组(P均<0.01);D组各时间点浓度均明显低于A组(P均<0.01)。4.RT PCR:IGFBP2mRNA表达的强度变化规律与其肽浓度变化相似。结论IGFBP2在肺发育过程起重要作用,其浓度过高则影响肺发育。
Objective To explore the role of insulin-like growth factor binding protein (IGFBP) 2 in lung development of neonatal rats. Methods Eighty SD pregnant rats were divided into 4 groups: A: control group; B: Dex group 1; C: Dex2 group; D: retinoic acid (RA) group. Rats in groups A, B and D were injected subcutaneously with normal saline at 18-20 d of gestation, Dex, and intraperitoneal injection of RA respectively. Group C was injected with Dex subcutaneously 1 ~ 3d after birth. Each group was taken at 18, 20 and 21 days of pregnancy (group C was not taken during pregnancy), and lung samples were taken for morphological examination at 1, 3, 5, 7, 10, 14 and 21 days after birth. Immunohistochemistry, Western blot and RT PCR . Lung tissue morphology: B, C group early alveolar development in advance, the number of more, wall thin, significantly lagged behind in the late A, D group. 2.IGFBP2 lung expression: IGFBP2 in group A was mainly expressed in fetal lung tissue, the expression was strongest on day 18, and then decreased gradually. The trend of expression in group B and C was the same as that in group A, but it was higher than group A at all time points after birth. The expression of IGFBP2 polypeptide in each group was the highest at 18d, then gradually decreased. The expression intensity at each time point in group B and C was significantly higher than that in group A at all time points (P <0.01) Significantly lower than group A (all P <0.01). RT PCR: The intensity of IGFBP2 mRNA expression changes with the changes of its peptide concentration similar. Conclusion IGFBP2 plays an important role in the process of lung development. The high concentration of IGFBP2 affects lung development.