Xu Wei, Xu Bo, Zhao Ying, Yang Ni, Liu Chunfeng, Wen Guangfu, Zhang Binglun
Department of Pediatrics, Shengjing Hospital of China Medical University, 36 Sanhao Street, Shenyang, Liaoning, 110004, People's Republic of China.
Department of Ophthalmology, The Affiliated People's Hospital of Jiangsu University, Zhenjiang, Jiangsu, 212002, People's Republic of China.
J Physiol Biochem. 2015 Dec;71(4):823-38. doi: 10.1007/s13105-015-0446-4. Epub 2015 Nov 7.
Transdifferentiation of alveolar epithelial type II cells (AECIIs) to type I cells (AECIs) is critical for reestablishment and maintenance of an intact alveolar epithelium. However, this process is frequently destroyed by hyperoxia treatment, which is commonly used in respiratory distress syndrome therapy in preterm infants. Wnt5a is considered to participate in this physiopathologic process, but the clear mechanisms still need to be further investigated. In this study, preterm rats and primary rat AECIIs were exposed to hyperoxia. Hematoxylin and eosin staining was used to examine the histological changes of the lungs. Real-time PCR and western blotting were used to examine Wnt5a expression and biomarkers of AECII and AECI expression. Immunohistochemistry and immunofluorescence were also used to determine the expression and location of selected biomarkers. Furthermore, AECIIs transfected with Wnt5a gene and exogenous Wnt5a were used to examine whether Wnt5a contributes to the transdifferentiation of AECIIs to AECIs. Results showed that hyperoxia inhibited the transdifferentiation of AECIIs to AECIs in vitro, which is represented by biomarkers of two types of cell that remained unchanged. In addition, Wnt5a protein expression was found to be decreased after hyperoxia exposure in vitro and in vivo. Furthermore, both the overexpression of Wnt5a and exogenous Wnt5a addition blocked the inhibitory effect of hyperoxia in vitro. In conclusion, our results suggest that the transdifferentiation of AECIIs to AECIs is impaired by hyperoxia, and this process may be associated with Wnt5a downregulation. Targeting Wnt5a may have the potential for the therapy of lung injury in preterm infants induced by hyperoxia.
肺泡Ⅱ型上皮细胞(AECIIs)向Ⅰ型细胞(AECIs)的转分化对于完整肺泡上皮的重建和维持至关重要。然而,这一过程经常因高氧治疗而被破坏,高氧治疗常用于早产儿呼吸窘迫综合征的治疗。Wnt5a被认为参与了这一病理生理过程,但其确切机制仍需进一步研究。在本研究中,将早产大鼠和原代大鼠AECIIs暴露于高氧环境。采用苏木精-伊红染色检查肺组织学变化。采用实时PCR和蛋白质印迹法检测Wnt5a表达以及AECII和AECI表达的生物标志物。免疫组织化学和免疫荧光也用于确定所选生物标志物的表达和定位。此外,用Wnt5a基因转染的AECIIs和外源性Wnt5a来研究Wnt5a是否有助于AECIIs向AECIs的转分化。结果显示,高氧在体外抑制了AECIIs向AECIs的转分化,这表现为两种类型细胞的生物标志物保持不变。此外,发现在体外和体内高氧暴露后Wnt5a蛋白表达均降低。此外,Wnt5a的过表达和外源性Wnt5a的添加均在体外阻断了高氧的抑制作用。总之,我们的结果表明,高氧损害了AECIIs向AECIs的转分化,这一过程可能与Wnt5a下调有关。靶向Wnt5a可能对治疗高氧诱导的早产儿肺损伤具有潜力。