Liu Xiaomei, Liu Caixia, Ma Tie, Jiao Yisheng, Miao Jianing, Gao Linlin
Department of Obstetrics and Gynecology, Shengjing Hospital of China Medical University, Shenyang, P. R. China.
Cell Physiol Biochem. 2016;39(1):385-94. doi: 10.1159/000445632. Epub 2016 Jul 4.
BACKGROUND/AIMS: Patients with esophageal atresia (EA) and tracheoesophageal fistula (TEF) often suffer chronic respiratory tract disease. We previously reported that primary lung maldevelopment caused by deficient branching of embryonal airways in experimental EA-TEF was induced by Adriamycin. In this study, we investigated the Vascular endothelial growth factor (VEGF) pathway in the developing lung in an EA-TEF rat model. We further analyzed the effect of recombinant VEGF treatment in vitro on branching morphogenesis of embryo lungs in experimental EA-TEF.
Pregnant rats received either Adriamycin or vehicle on E7, E8 and E9. Lungs were recovered at E15, E18 and E21. Expression of VEGF and receptors (Flk-1 and Flt-1) were assessed by quantitative PCR, immunohistochemistry and immunoblotting. E13 lungs were cultured for 72 hours with 50 ng/mL of recombinant rat VEGF in serum-free medium. The rates of increase in bud count and airway contour were evaluated.
Our results showed a significant downregulation of VEGF during pseudoglandular and canalicular stages. In contrast, there were significantly higher levels of the Flt-1 receptor in the canalicular stage, which may represent a compensatory response to decreased VEGF. However, both variables returned to normal levels at the saccular stage. Exogenous VEGF treatment enhanced hypoplastic lung growth, evidenced by the increase in bud count and airway contour.
A VEGF signaling defect possibly plays an important role in defective embryonic airway branching. Additionally, VEGF treatment may accelerate lung growth in EA-TEF lungs.
背景/目的:食管闭锁(EA)和气管食管瘘(TEF)患者常患有慢性呼吸道疾病。我们之前报道过,阿霉素可诱导实验性EA - TEF中胚胎气道分支缺陷导致的原发性肺发育不良。在本研究中,我们在EA - TEF大鼠模型中研究了发育中肺的血管内皮生长因子(VEGF)通路。我们进一步分析了体外重组VEGF治疗对实验性EA - TEF中胚胎肺分支形态发生的影响。
怀孕大鼠在胚胎第7天、第8天和第9天接受阿霉素或赋形剂处理。在胚胎第15天、第18天和第21天取出肺组织。通过定量PCR、免疫组织化学和免疫印迹法评估VEGF及其受体(Flk - 1和Flt - 1)的表达。将胚胎第13天的肺组织在无血清培养基中用50 ng/mL重组大鼠VEGF培养72小时。评估芽计数和气道轮廓的增加率。
我们的结果显示,在假腺期和小管期VEGF显著下调。相比之下,小管期Flt - 1受体水平显著升高,这可能代表对VEGF减少的一种代偿反应。然而,在囊状期这两个变量均恢复到正常水平。外源性VEGF治疗可促进发育不全的肺生长,芽计数和气道轮廓增加证明了这一点。
VEGF信号缺陷可能在胚胎气道分支缺陷中起重要作用。此外,VEGF治疗可能加速EA - TEF肺的生长。