AAV delivery of tumor necrosis factor-α short hairpin RNA attenuates cold-induced pulmonary hypertension and pulmonary arterial remodeling.
AAV delivery of tumor necrosis factor-α short hairpin RNA attenuates cold-induced pulmonary hypertension and pulmonary arterial remodeling.
复制标题
DOI：
10.1161/hypertensionaha.114.03791
复制
发表时间：
2014-11
期刊：
影响因子：
--
通讯作者：
Sun Z
中科院分区：
文献类型：
--
作者：
Crosswhite P;Chen K;Sun Z
低温与心血管和肺部疾病的死亡率和发病率增加有关。冷暴露可引起肺部炎症、肺动脉高压（PH）和右心室（RV）肥大，但由于机制不明，目前尚无有效的治疗方法。在这里，我们研究了TNF-α的RNAi沉默是否减少冷诱导的巨噬细胞浸润，PH和肺动脉（PA）重塑。我们首次发现，在RV收缩压升高之前，连续冷暴露（5.0°C）增加了肺和肺动脉（PA）中TNF-α的表达和巨噬细胞浸润。通过在冷暴露前24小时IV递送携带TNFα短发夹siRNA（TNFshRNA）的重组AAV-2来实现TNF-α的体内RNAi沉默。与温暖对照组相比，冷暴露8周显著增加RV压力（40.19±4.9 vs 22.9±1.1 mmHg），表明冷暴露导致PH。冷暴露增加肺和PA中TNF-α、IL-6和磷酸二酯酶-1C（PDE-1C）蛋白表达，并增加肺巨噬细胞浸润。值得注意的是，TNFshRNA阻止了冷诱导的TNF-α、IL-6和PDE-1C蛋白表达的增加，消除了肺巨噬细胞浸润，并减轻了PH（26.28±1.6 mmHg）、PA重塑和RV肥大。从冷暴露动物分离的PA SMC显示出细胞内超氧化物水平增加和细胞增殖沿着细胞内cGMP降低。这些冷诱导的变化被TNFshRNA阻止。TNF-α的上调通过促进肺巨噬细胞浸润和炎症反应在冷诱导PH的发病机制中起关键作用。AAV递送TNFshRNA可能是冷诱导的PH和PA重构的有效治疗方法。
Cold temperatures are associated with increased mortality and morbidity of cardiovascular and pulmonary disease. Cold exposure causes lung inflammation, pulmonary hypertension (PH) and right ventricle (RV) hypertrophy, but there is no effective therapy due to unknown mechanism. Here we investigated if RNAi silencing of TNF-α decreases cold-induced macrophage infiltration, PH, and pulmonary arterial (PA) remodeling. We found for the first time that continuous cold exposure (5.0°C) increased TNF-α expression and macrophage infiltration in the lungs and pulmonary arteries (PAs) right before elevation of RV systolic pressure. The in vivo RNAi silencing of TNF-α was achieved by IV delivery of recombinant AAV-2 carrying TNFα short hairpin siRNA (TNFshRNA) 24 hours prior to cold exposure. Cold exposure for eight weeks significantly increased RV pressure compared to the warm controls (40.19±4.9 vs. 22.9±1.1 mmHg), indicating that cold exposure caused PH. Cold exposure increased TNF-α, IL-6 and phosphodierterase-1C (PDE-1C) protein expression in the lungs and PAs and increased lung macrophage infiltration. Notably, TNFshRNA prevented the cold-induced increases in TNF-α, IL-6 and PDE-1C protein expression, abolished lung macrophage infiltration, and attenuated PH (26.28±1.6 mmHg), PA remodeling, and RV hypertrophy. PA SMCs isolated from cold-exposed animals showed increased intracellular superoxide levels and cell proliferation along with decreased intracellular cGMP. These cold-induced changes were prevented by TNFshRNA. Upregulation of TNF-α played a critical role in the pathogenesis of cold-induced PH by promoting pulmonary macrophage infiltration and inflammation. AAV delivery of TNFshRNA may be an effective therapeutic approach for cold-induced PH and PA remodeling.