Electroporation-mediated in vivo gene delivery of the Na+/K+-ATPase pump reduced lung injury in a mouse model of lung contusion.
Electroporation-mediated in vivo gene delivery of the Na+/K+-ATPase pump reduced lung injury in a mouse model of lung contusion.
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DOI：
10.1097/ta.0b013e31823f0606
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发表时间：
2012-01
期刊：
影响因子：
--
通讯作者：
Raghavendran K
中科院分区：
文献类型：
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作者：
Machado-Aranda DA;Suresh MV;Yu B;Raghavendran K
肺挫伤(LC)是成人呼吸窘迫综合征(ARDS)的独立危险因素。ARDS的最后一个共同途径是肺泡积液。在这项研究中，我们展示了一种潜在的基因治疗方法的应用，通过递送Na+/K+-ATPase泵亚单位在小鼠LC模型中。我们假设，恢复泵的活性将导致清除多余的肺泡液，并另外减少炎症。C57/BL6小鼠在麻醉状态下，用皮质挫伤冲击器沿肋缘上方1 cm处的右腋后线撞击。即刻将编码Na+/K+-ATPase泵α，β的100 mgDNA注入肺内(LC EP泵组)。对照组为单纯挫伤组(LC)和伤后假生理盐水组(LC EP-Sham)。使用BTX 830型电穿孔机，经胸腔内施加8个场强为200V/cm的电脉冲。分别于产后24小时、48小时和72小时处死小鼠。收集支气管肺泡灌洗液(BAL)，用ELISA法测定白蛋白和细胞因子。测量肺顺应性，并进行肺组织病理学分析。在电穿孔和运送编码Na+/K+-ATPase泵α，β亚单位的基因后，急性肺损伤显著减轻，表现为白蛋白水平降低，P-V曲线改善，组织学炎症减轻。电穿孔介导的Na+/K+-ATPase泵亚单位基因转移促进了LC后急性炎症性肺损伤的恢复。
Lung contusion (LC) is an independent risk factor for Adult Respiratory Distress Syndrome (ARDS). The final common pathway in ARDS involves accumulation of fluid in the alveoli. In this study we demonstrate the application of a potential gene therapy approach by delivering the Na+/K+-ATPase pump sub-units in a murine model of LC. We hypothesized that restoring the activity of the pump will result in removal of excess alveolar fluid and additionally reduce inflammation. Under anesthesia, C57/BL6 mice were struck along the right posterior axillary line 1 cm above the costal margin with a cortical contusion impactor. Immediately afterwards; 100 μg of plasmid DNA coding for the α,β of the Na+/K+-ATPase pump were instilled into the lungs (LC EP-pump group). Contusion only (LC only) and a sham saline instillation group after contusion were used as controls (LC EP-sham). Using a BTX 830 Electroporator, 8 electrical pulses of 200-V/cm field strength were applied transthoracically. Mice were sacrificed at 24hr, 48hr and 72hr post-delivery. Bronchial alveolar lavage (BAL) was recollected to measure albumin and cytokines by ELISA. Pulmonary compliance was measured and lungs were subject to histopathologic analysis. Following the electroporation and delivery of genes coding for the α,β subunits of the Na+/K+-ATPase pump, there was a significant mitigation of acute lung injury as evidenced by reduction in BAL levels of albumin, improved P-V curves and reduced inflammation seen on histology. Electroporation mediated gene transfer of the subunits of the Na+/K+-ATPase pump enhanced recovery from acute inflammatory lung injury following LC.