β1-Na+,K+-ATPase gene therapy upregulates tight junctions to rescue lipopolysaccharide-induced acute lung injury
β1-Na+,K+-ATPase gene therapy upregulates tight junctions to rescue lipopolysaccharide-induced acute lung injury
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DOI：
10.1038/gt.2016.19
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发表时间：
2016-06-01
期刊：
影响因子：
5.1
通讯作者：
Dean, D. A.
中科院分区：
文献类型：
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作者：
Lin, X.;Barravecchia, M.;Dean, D. A.
急性肺损伤（ALI）和急性呼吸窘迫综合征（ARDS）与多种疾病相关，其特征是肺泡-毛细血管屏障破坏，水肿液渗漏到肺部，严重炎症导致急性呼吸衰竭。基因治疗是通过修复肺泡上皮功能来治疗ALI/ARDS的潜在有效途径。本研究表明，通过电穿孔将表达Na+， K+- atp酶β 1亚基（β 1-Na+， K+- atp酶）的质粒单独或与上皮钠通道(ENaC) α 1亚基联合递送，不仅可以保护随后的脂多糖（LPS）介导的肺损伤，还可以治疗损伤的肺。然而，单独转移ENaC的α 1亚单位（α 1-ENaC）仅提供保护效益，而不是治疗效益，尽管肺泡液清除率显着提高。基因转移β 1-Na+、K+-ATPase，而非α 1-ENaC，不仅在培养细胞和小鼠肺中增强了紧密连接蛋白-1 （ZO-1）和occludin的表达，而且在体内降低了原有的肺通透性增加。这些结果表明，β 1-Na+， K+-ATPase的基因转移上调紧密连接的形成，从而治疗已有损伤的肺，而α 1-ENaC的传递仅维持原有的紧密连接，而不是产生。这表明上皮/内皮屏障功能的恢复可能为ALI/ARDS提供更好的治疗。
Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are associated with diverse disorders and characterized by disruption of the alveolar-capillary barrier, leakage of edema fluid into the lung, and substantial inflammation leading to acute respiratory failure. Gene therapy is a potentially powerful approach to treat ALI/ARDS through repair of alveolar epithelial function. Herein, we show that delivery of a plasmid expressing beta 1-subunit of the Na+, K+-ATPase (beta 1-Na+, K+-ATPase) alone or in combination with epithelial sodium channel (ENaC) alpha 1-subunit using electroporation not only protected from subsequent lipopolysaccharide (LPS)-mediated lung injury, but also treated injured lungs. However, transfer of alpha 1-subunit of ENaC (alpha 1-ENaC) alone only provided protection benefit rather than treatment benefit although alveolar fluid clearance had been remarkably enhanced. Gene transfer of beta 1-Na+, K+-ATPase, but not alpha 1-ENaC, not only enhanced expression of tight junction protein zona occludins-1 (ZO-1) and occludin both in cultured cells and in mouse lungs, but also reduced pre-existing increase of lung permeability in vivo. These results demonstrate that gene transfer of beta 1-Na+, K+-ATPase upregulates tight junction formation and therefore treats lungs with existing injury, whereas delivery of alpha 1-ENaC only maintains pre-existing tight junction but not for generation. This indicates that the restoration of epithelial/endothelial barrier function may provide better treatment of ALI/ARDS.