1 Department of Neonatology, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing, China.
2 China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Children's Hospital of Chongqing Medical University, Chongqing, China.
J Interferon Cytokine Res. 2019 Feb;39(2):106-116. doi: 10.1089/jir.2018.0080. Epub 2019 Jan 18.
Abnormal alveolar formation and excessive disordered elastin accumulation are key pathological features in bronchopulmonary dysplasia. Transforming growth factor (TGF)-β is an important regulator of the extracellular matrix in the developing lung. To determine if increased TGF-β would injure alveolar development by activating TGF-β signaling and by influencing the expression of elastogenesis-related protein, we performed intraperitoneal injection of newborn mice with the TGF-β-neutralizing antibody 1D11 and observed whether 1D11 had a protective role in the oxygen (O)-exposed newborn mouse lung. The newborn mice were exposed to 85% O for 14 and 21 days. 1D11 was administered by intraperitoneal injection every day from postnatal days 3 to 20. Alveolar morphology was assessed by hematoxylin and eosin staining. The expression and distribution of elastin were evaluated by immunohistochemistry. The level of TGF-β signaling-related proteins were measured by immunohistochemistry, enzyme-linked immunosorbent assay, and Western blot. The expression levels of elastogenesis-related proteins, including tropoelastin, fibulin-5, and neutrophil elastase (NE), which participate in the synthesis, assembly, and degradation of elastin, were detected by real-time PCR and Western blot. In this research, impaired alveolar development and elastin deposition as well as the excessive activation of TGF-β signaling were observed in the newborn mouse lung exposed to hyperoxia. 1D11 improved alveolarization as well as the distribution of elastin in the newborn lung with hyperoxia exposure. The expression levels of tropoelastin, fibulin-5, and NE, which are important components of elastogenesis, were decreased by treatment with 1D11 in the injured newborn lung. These data demonstrate that 1D11 improved alveolarization by blocking the TGF-β signaling pathway and by reducing the abnormal expression of elastogenesis-related proteins in the O-exposed newborn mouse lung. 1D11 may become a new therapeutic method to prevent the development of bronchopulmonary dysplasia.
异常的肺泡形成和过度紊乱的弹性蛋白积累是支气管肺发育不良的关键病理特征。转化生长因子 (TGF)-β 是发育中肺细胞外基质的重要调节剂。为了确定增加 TGF-β 是否通过激活 TGF-β 信号通路和影响弹性蛋白生成相关蛋白的表达来损伤肺泡发育，我们用 TGF-β 中和抗体 1D11 对新生小鼠进行腹腔注射，并观察 1D11 在氧（O）暴露的新生鼠肺中是否具有保护作用。新生小鼠暴露于 85% O 中 14 和 21 天。1D11 从出生后第 3 天至第 20 天每天通过腹腔注射给药。通过苏木精和伊红染色评估肺泡形态。通过免疫组织化学评估弹性蛋白的表达和分布。通过免疫组织化学、酶联免疫吸附试验和 Western blot 测量 TGF-β 信号相关蛋白的水平。通过实时 PCR 和 Western blot 检测参与弹性蛋白合成、组装和降解的弹性蛋白生成相关蛋白，包括原肌球蛋白、纤维连接蛋白-5 和中性粒细胞弹性蛋白酶（NE）的表达水平。在这项研究中，在高氧暴露的新生鼠肺中观察到肺泡发育受损和弹性蛋白沉积以及 TGF-β 信号的过度激活。1D11 改善了高氧暴露新生鼠肺的肺泡化和弹性蛋白分布。用 1D11 处理可降低损伤新生肺中弹性蛋白生成相关蛋白原肌球蛋白、纤维连接蛋白-5 和 NE 的表达水平。这些数据表明，1D11 通过阻断 TGF-β 信号通路并减少 O 暴露新生鼠肺中弹性蛋白生成相关蛋白的异常表达来改善肺泡化。1D11 可能成为预防支气管肺发育不良的新治疗方法。
