Weng Y H, Tatarov A, Bartos B P, Contag C H, Dennery P A
Department of Pediatrics, Stanford University School of Medicine, Palo Alto, California 94304, USA.
Am J Physiol Lung Cell Mol Physiol. 2000 Jun;278(6):L1273-9. doi: 10.1152/ajplung.2000.278.6.L1273.
Somatic cell gene transfer is a potentially useful strategy to alter lung function. However, achieving efficient transfer to the alveolar epithelium, especially in smaller animals, has not been demonstrated. In this study, the rat heme oxygenase-1 (HO-1) gene was delivered to the lungs of neonatal mice via transpulmonary injection. A bidirectional promoter construct coexpressing both HO-1 and a luciferase reporter gene was used so that in vivo gene expression patterns could be monitored in real time. HO-1 expression levels were also modulated with doxycycline and assessed in vivo with bioluminescent light transmitted through the tissues from the coregulated luciferase reporter. As a model of oxidative stress and HO-1-mediated protection, groups of animals were exposed to hyperoxia. After gene transfer, elevated levels of HO-1 were detected predominantly in alveolar type II cells by immunocytochemistry. With overexpression of HO-1, increased oxidative injury was observed. Furthermore, this model demonstrated a cell-specific effect of lung HO-1 overexpression in oxidative stress. Specific control of expression for therapeutic genes is possible in vivo. The transpulmonary approach may prove useful in targeting gene expression to cells of the alveolar epithelium or to circumscribed areas of the lung.
体细胞基因转移是一种改变肺功能的潜在有用策略。然而，尚未证明能有效地将基因转移到肺泡上皮，尤其是在较小的动物中。在本研究中，通过经肺注射将大鼠血红素加氧酶-1（HO-1）基因导入新生小鼠的肺中。使用了一种共表达HO-1和荧光素酶报告基因的双向启动子构建体，以便实时监测体内基因表达模式。HO-1表达水平也用强力霉素进行调节，并通过共调节的荧光素酶报告基因透过组织发出的生物发光在体内进行评估。作为氧化应激和HO-1介导的保护作用的模型，将动物分组暴露于高氧环境。基因转移后，通过免疫细胞化学检测到HO-1水平升高主要出现在肺泡II型细胞中。随着HO-1的过表达，观察到氧化损伤增加。此外，该模型证明了肺HO-1过表达在氧化应激中的细胞特异性作用。在体内对治疗性基因进行特异性表达控制是可能的。经肺途径可能被证明对将基因表达靶向肺泡上皮细胞或肺的特定区域有用。
